KR20100108544A - Vaccine - Google Patents

Abstract

The present invention provides a replication defect that encodes a protein comprising a CS protein derived from P. falciparum or a fragment thereof (eg, as shown in Seq ID No: 1 or Seq ID No: 3). And simian adenovirus vector C7. The present invention further relates to a method of producing said viral vector and to the use of said viral vector in the treatment / prevention of malaria infection. Also described are compositions, vaccines and kits comprising such viral vectors. In a first aspect, the invention uses a synthetic C7 virus vector. C7 viral vectors according to the invention are co-administered or co-formulated with malaria antigens such as RTS, S, optionally in the presence of an adjuvant comprising, eg, 3D-MPL and / or saponins such as QS21 Can be converted.

Description

Vaccine {VACCINE}

The present invention relates to simian-derived adenovirus vectors that specifically encode novel malaria antigens derived from circumsporozoite proteins of Plasmodium falciparum . The invention further relates to methods of making said viral vectors and to the use of said viral vectors in the treatment / prevention of malaria infections.

Malaria, one of the world's major health problems, kills between two and four million people every year.

One of the most acute form of the disease is a protozoan parasite that causes the majority of deaths due to malaria, falciparum parasite (Plasmodium falciparum ) ( P. falciparum ).

The life cycle of P. falciparum is complex and requires two hosts, humans and mosquitoes, to complete this cycle. Human infection begins by inoculation of sporozoites in saliva as a bite of an infected mosquito. Sporozoites migrate to the liver to infect hepatocytes, from which they undergo extracellular extracellular erythrocytes to differentiation into the mesozoite phase that infects red blood cells (RBCs), which initiate circulating replication in the asexual blood phase. The circulation is completed by differentiating into the sexual stage germ cells of a number of merozoids in RBCs, then being eaten by mosquitoes, developing through successive periods of the mid- to long-term, producing sporozoites and then moving to salivary glands.

The sporozoite phase of Plasmodium has been identified as a potential target for malaria vaccines. Vaccination with inactivated (irradiated) sporozoite has been shown to induce protection against experimental human malaria (Am. J, Trap. Med. Hyg 24: 297-402, 1975). However, based on this methodology, using sporozoites investigated, it was practically and logically impossible to produce vaccines against malaria for the masses.

Sporozoite's major surface protein is known as sulfophosphozyte protein (CS protein). In its migration from the initial site of inoculation by mosquitoes into the circulation (delivered to the liver), it is believed that the motility and invasiveness of the sporozoite are involved.

CS proteins of Plasmodia species are characterized by a central repeating domain (repeat region) next to non-repeating amino (N-terminus) and carboxy (C-terminus) fragments.

To date, the most advanced malaria vaccines in hospitals are based on lipoprotein particles (also known as virus-like particles) called RTS, S. The particles are substantially fused to the N- terminus of the S antigen derived from Hepatitis B Plasmodium falciparum CS protein of the amino acid (P. falciparum) (NF54 [3D7 ] system) 207-395DP corresponding to, P. falciparum (P. falciparum ) portion of the CS protein. The S antigen may comprise a portion of preS2.

RTS and S particles are usually delivered with a strong adjuvant.

Although malaria vaccines have been proposed to use recombinant adenovirus vectors (eg, WO 2004/055187 describes specific viral vectors comprising specific adeno 5 (Ad5) and adeno 35 (Ad 35) vectors). Both were derived from human adenovirus encoding the CS protein.

There are more than 40 different serotypes of human adenoviruses, and their pathogenicity varies, eg Ad5 is associated with mild respiratory infections in children, Ad4 and Ad7 are believed to be involved in adult respiratory infections, and Ad40 Is thought to cause diarrhea in infants.

Immunity of adenovirus infections is thought to last a lifetime after infection. In particular, pre-existing immunity to Ad5 and Ad35 results in neutralization of therapeutic adenovirus vectors based on human adenovirus. This can reduce the therapeutic effectiveness of the vector because it prevents the vector from entering the cell and producing the proper antigen in the body.

The present invention is believed to reduce the problem of existing immunity by providing a vaccine for the prevention and / or treatment of malaria, comprising: a protein comprising a CS protein derived from P. falciparum or its Replication defects encoding fragments appear in Simian Adenovirus Vector C7 (also referred to as Pan 7 or CV-33) such as Seq ID No: 1 or Seq ID No: 3.

Brief description of the sequence

Seq ID No: 1 falciparum (referred to herein as Ade2 protein) amino acid sequence of a protein / antigen derived from CS protein falciparum (P.falciparum).

Seq ID No: 2 Nucleic acid sequence encoding a protein of Seq ID No: 1, referred to herein as the Ade2 gene.

Seq ID No: 3 Alternative amino acid sequence of protein / antigen derived from CS protein of P. falciparum (herein referred to as Ade1 protein).

Seq ID No: 4 Nucleic acid sequence encoding a protein of Seq ID No: 3, referred to herein as the Ade1 gene.

Seq ID No: 5 Capsid protein sequence from Chimp Adeno 7 (seq ID No 17 from WO 03/046124).

Seq ID No: 6 amino acid sequence from chimpanzee adeno 7 (seq ID No 20 from WO 03/046124).

Seq ID No: 7 amino acid sequence from P. falciparum CS protein.

Seq ID No: 8 amino acid sequence from P. falciparum CS protein.

Seq ID No: 9 amino acid sequence from P. falciparum CS protein.

Seq ID No: 10 amino acid sequence from P. falciparum CS protein.

Nucleotide sequence for Seq ID No: 11 CpG 1826.

Seq ID No: nucleotide sequence for 12 CpG 1758.

Seq ID No: nucleotide sequence for 13 CpG.

Nucleotide sequence for Seq ID No: 14 CpG 2006.

Seq ID No: nucleotide sequence for 15 CpG 1668.

Seq ID No: nucleotide sequence for 16 CpG 5456.

Seq ID No: 17 shows nucleotide sequence of alternative expression cassette for Ade2 expression cassette (cloned into C7 adenovirus vector).

Seq ID No: shows nucleotide sequence of 18 Ade2 expression cassette (replicated with C7 adenovirus vector).

Seq ID No: 19 shows the complete nucleotide sequence of the synthetic recombinant vector C7 -Ade2.

Brief description of the drawings

1 shows a plasmid map for pCR2.1-Ade2.

2 shows the plasmid map for pShuttle6-Ade2.

3 shows the plasmid map for pC7000-CMV Ade2.

4-7 show comparisons between CS-specific T cell responses induced by C7 Ade1 and C7 Ade2 in C57B1 / 6 mice.

8-11 show a comparison between CS-specific T cell responses induced by C7 Ade2 and Ad5 Ade2 in C57B1 / 6 mice.

12-15 show a comparison between CS-specific T cell responses induced by C7 Ade1 and Ad5 Ade1 in C57B1 / 6 mice.

16 depicts anti-CS antibody response measured by ELISA in C57B1 / 6 mice.

17 and 18 show the kinetics of CS-specific CD8 T cell responses induced by C7-Ade2 in CB6F1 mice.

19 and 20 show the kinetics of CS-specific CD4 T cell responses induced by C7-Ade2 in CB6F1 mice.

21 and 22 show cytokine profiles of CS-specific CD8 T cell responses induced by C7-Ade2 in CB6F1 mice.

Figures 23 and 24 show cytokine profiles of CS-specific CD4 T cell responses induced by C7-Ade2 in CB6F1 mice.

25 and 26 show the kinetics of CS-specific CD8 T cell responses co-formulated with RTS, S / AS01B in CB6F1 mice or induced by C7-Ade2 in a prime / boost manner It is shown.

27 and 28 show the kinetics of CS-specific CD4 T cell responses co-formulated with RTS, S / AS01B in CB6F1 mice or induced by C7-Ade2 in a prime / boost manner.

FIG. 29 shows the kinetics of HBs-specific CD8 T cell responses co-formulated with RTS, S / AS01B in CB6F1 mice or induced by C7-Ade2 in a prime / boost manner.

30 depicts kinetics of HBs-specific CD4 T cell responses co-formulated with RTS, S / AS01B in CB6F1 mice or induced by C7-Ade2 in a prime / boost manner.

31 and 32 show cytokine profiles of CS-specific CD8 T cell responses co-formulated with RTS, S / AS01B in CB6F1 mice or induced by C7-Ade2 in a prime / boost manner.

Figure 33 depicts cytokine profiles of HBs-specific CD8 T cell responses co-formulated with RTS, S / AS01B in CB6F1 mice or induced by C7-Ade2 in a prime / boost manner.

34 and 35 show cytokine profiles of CS-specific CD4 T cell responses co-formulated with RTS, S / AS01B in CB6F1 mice or induced by C7-Ade2 in a prime / boost manner. .

FIG. 36 shows cytokine profiles of HBs-specific CD4 T cell responses co-formulated with RTS, S / AS01B in CB6F1 mice or induced by C7-Ade2 in a prime / boost manner.

37 and 38 show antibody responses co-formulated with RTS, S / AS01B in CB6F1 mice or induced by C7-Ade2 in a prime / boost manner.

39-41 show the kinetics of CS- and HBs-specific CD8 T cell responses induced by co-formulated C7-Ade2 + RTS, S / AS01B in CB6F1 mice.

42-44 depict the kinetics of CS- and HBs-specific CD4 T cell responses induced by co-formulated C7-Ade2 + RTS, S / AS01B in CB6F1 mice.

45-47 show cytokine profiles of CS- and HBs-specific CD8 T cell responses induced by co-formulated C7-Ade2 + RTS, S / AS01B in CB6F1 mice.

48-50 show cytokine profiles of CS- and HBs-specific CD4 T cell responses induced by co-formulated C7-Ade2 + RTS, S / AS01B in CB6F1 mice.

51 and 52 show antibody responses induced by C7-Ade2 + RTS, S / AS01B co-formulated in CB6F1 mice.

53-55 show the kinetics of CS- and HBs-specific CD8 T cell responses induced by C7-Ade2, RTS, S and ASOlB responses in CB6F1 mice.

56-58 show the kinetics of CS- and HBs-specific CD4 T cell responses induced by C7-Ade2, RTS, S and ASOlB responses in CB6F1 mice.

59-61 show cytokine profiles of CS- and HBs-specific CD8 T cell responses induced by C7-Ade2, RTS, S and ASOlB responses in CB6F1 mice.

62-64 show cytokine profiles of CS- and HBs-specific CD4 T cell responses induced by C7-Ade2, RTS, S and ASOlB responses in CB6F1 mice.

65 and 66 show antibody responses induced by C7-Ade2, RTS, S and ASOlB responses in CB6F1 mice.

67 and 68 show the kinetics of CS-specific CD8 T cell responses induced by synthetic C7 Ade2 in CB6F1 mice.

69 and 70 depict the kinetics of CS-specific CD4 T cell responses induced by synthetic C7 Ade2 in CB6F1 mice.

71 and 72 show cytokine profiles of CS-specific CD8 T cell responses induced by synthetic C7 Ade2 in CB6F1 mice.

73 and 74 depict cytokine profiles of CS-specific CD4 T cell responses induced by synthetic C7 Ade2 in CB6F1 mice.

The sequence and preparation of C7 is described in WO 2003/046124. SEQ ID Nos: 6 (Fenton sequence), 9 (nucleic acid sequence), 10 and 11 (hexon sequence), and 12 (fiber protein) of WO 2003/046124 are incorporated by reference. The registration number for C7 is [ATCC VR-593].

Although the hypothesis exists that vectors can be grouped into families, and that adenovirus vectors within a given family may have similar characteristics, the features and characteristics of any given adenovirus vector are largely individual.

It is considered particularly advantageous to use C7 because C7 will be more stable than certain other known vectors, such as C6 disclosed in WO 2003/046124, once the protein encoding the gene is inserted. This means that C7 is less prone to re-organisation. Of course, it is very important that any adenovirus vectors used in the vaccines are stable, because they need to be proven and well characterized, before they can be marketed.

Existing immunity to C7 is believed to be very low and therefore the risk of neutralization of the viral vector after the initial administration to the patient is low.

In addition, it is believed that one or more properties of C7 exist, which makes it particularly suitable to produce a favorable immune response in administration and / or in vivo.

In a first aspect, the present invention uses synthetic C7 viral vectors, which may be particularly suitable for obtaining regulatory approval for administration to humans.

In a first embodiment, the malaria antigen component derived from the CS protein has been removed from the last 12-14 amino acids.

In a first embodiment, the malaria antigen encoded by the adenovirus vector is modified to remove potential glycosylation sites, such as amino acid alanine can be replaced with serine (eg, at about 379 positions of Seq ID No: 1). As shown).

In a first aspect, the protein / antigen used in the present invention comprises the following amino acids:

Optionally located at about amino acids 81-99.

In a first embodiment, the protein / antigen used comprises the following amino acids:

Eg located at the C terminus.

In a first aspect, the invention uses a protein comprising the following amino acids:

In a first aspect, the invention uses a protein comprising the following amino acids:

In further embodiments, the protein / antigen used is selected from Seq ID No. 1. 7 and / or Seq ID No. 8 and / or the sequence of Seq ID No 9;

In further embodiments, the protein / antigen used is selected from Seq ID No. 1. 7 and / or Seq ID No. 8 and / or the sequence of Seq ID No 10.

In a first embodiment, the encoded protein / antigen is selected from Seq ID No. 1. 1 or 3.

The protein sequence given in Seq ID No: 1 is novel and forms an aspect of the present invention.

In addition, polynucleotides encoding the protein sequence of Seq ID No: l, in particular the polynucleotide sequence of Seq ID No: 2, form an aspect of the present invention. This polynucleotide sequence (ID No: 2) is already codon-optimized for expression in humans.

Optionally, the polynucleotide sequence encoding the protein of Seq ID No: 1 can be codon-optimized.

The invention also extends to the vectors / plasmids / hosts used in the preparation of novel hybrid fusion proteins of Seq ID No: 1 or in the preparation of viral vectors according to the invention.

When the preparation and isolation of a protein is required, a suitable plasmid can be used to insert the sequence encoding the protein into a suitable host for synthesis. Examples of suitable plasmids are pRIT 15546, 2 micron-based vectors for delivering suitable expression cassettes. The plasmid will generally contain intrinsic markers to aid selection, such as antibiotic resistance or genes encoding for LEU2 or HIS auxotrophy.

The host cell may be a prokaryotic or progressive cell, but preferably, yeasts such as Saccharomyces (eg Saccharomyces cerevisiae , such as DC5 (Accession No. 20820) under the name RIT DC5 cir (o) in the ATCC database. Trustee Smith Kline-RIT) and non - Saccharomyces yeast. This ski irradiation Caro My process (Schizosaccharomyces) (e.g., ski irradiation Caro My process pombe (Schizosaccharomyces pombe)), Cluj Vero My process (Kluyveromyces) (e. G., Vero Cluj My process lactis (Kluyveromyces lactis)), blood teeth (Pichia) (for example, pitch APA storage switch (Pichiapastoris)), a century Cronulla (Hansenula) (e.g., poly century Cronulla amorphastilbol (Hansenula polymorpha )) , Yarrowia (e.g., Yarrowia lipolytica)) and ski Wani Oh, my process (Schwanniomyces) (e.g., ski Wani Oh, my process come denta less (Schwanniomyces occidentalis )).

In a first aspect, the invention provides a protein of Seq ID No 1 for the use of the vector according to the invention or for the treatment or prevention of malaria.

In a first aspect the invention provides a pharmaceutical formulation comprising a viral vector according to the invention and an excipient such as an isotonic carrier suitable for infusion. Suitable excipients are discussed in more detail below.

In a first embodiment, the formulation comprises:

Adenovirus vectors according to the invention,

Malaria antigens such as lipoprotein particles, in particular RTS, S, and

Optionally including an adjuvant such as saponin and / or 3D-MPL.

If the vector encodes the sequence of Seq ID No: 1, the vector is particularly suitable for use in therapeutic regimens with proteins known as RTS, S. This is because the protein encoded by the adenovirus vector corresponds as closely as possible to the "RT" component of RTS, S. The use of the vector in regimens with RTS, S is thought to have the ability to effectively enhance the efficacy of RTS, S.

The viral vectors described herein are suitable for use as components for malaria vaccines. The viral vectors of the invention will need to be used in combination with other components, including other antigens, to provide adequate protection against infection. Nevertheless, the vectors of the invention are suitable for use at least as a component of a vaccine or in a therapeutic regimen.

RTS , S

RTS, S can be prepared as described in WO 93/10152 (eg from the P. falciparum NF54 / 3D7 strain). Nucleotide sequences for the RTS expression cassette and expected translation products are provided in FIG. 9 of WO 93/10152 (referred to herein as RTS *).

In the description of the invention, an excipient refers to one component in a pharmaceutical formulation that does not have a therapeutic effect by itself. Diluents or carriers are included within the definition of excipients. Suitable carriers include PBS, saline and the like. Adjuvants are also included within the definition of excipients, since the adjuvant may have a physiological effect in vivo, but this effect is common and in the absence of a therapeutic component does not exhibit a particular therapeutic effect.

Adjuvant

Certain adjuvants are metal salts, oil in water emulsions, toll like receptors agonists (especially toll like receptor 2 agonists, toll like receptor 3 agonists, toll like receptor 4 efficacy Agent, toll like receptor 7 agonist, toll like receptor 8 agonist, toll like receptor 9 agonist), saponin, or a combination thereof.

In a first embodiment, the adjuvant is a toll like receptor (TLR) 4 ligand, such as an agonist such as a lipid A derivative, in particular monophosphoryl lipid A or more particularly 3-diacylated monophosphoryl lipid A (3D-MPL). )to be.

3-Diacylated monophosphoryl lipid A is known from US Pat. No. 4,912,094 and UK Patent Application No. 2,220,211 (Ribi) and is available from Ribi Immunochem, Montana, USA.

3D-MPL is marketed under the trademark MPL® by Corixa Corporation and promotes CD4 + T cell responses with a predominantly IFN-g (ThI) phenotype. 3D-MPL can be produced according to the method described in GB 2 220 211 A. Chemically, 3D-MPL is a mixture of 3-diacylated monophosphoryl lipid A with 3, 4, 5 acylated chains. Generally, in the compositions of the present invention, small particle 3D-MPL is used. Small particle 3D-MPL has a particle size that can be sterile filtered through a 0.22 μm filter. Such preparations are described in WO 94/21292. Synthetic derivatives of lipid A are known and are believed to be TLR 4 agonists, including but not limited to:

OM174 (2-deoxy-6-O- [2-deoxy-2-[(R) -3-dodecanoyloxytetra-decanoylamino] -4-o-phosphono-β-D-glucopyra Nosyl] -2-[(R) -3-hydroxytetradecanoylamino] -α-D-glucopyranosyldihydrogenphosphate) (WO 95/14026),

OM 294 DP (3S, 9R) -3-[(R) -dodecanoyloxytetradecanoylamino] -4-oxo-5-aza-9 (R)-[(R) -3-hydroxytetradeca Noylamino] decane-1,10-diol, 1,10-bis (dihydrogenophosphate) (WO 99/64301 and WO 00/0462),

OM 197 MP-Ac DP (3S-, 9R) -3-[(R) -dodecanoyloxytetradecanoylamino] -4-oxo-5-aza-9-[(R) -3-hydroxytetra Decanoylamino] decane-1,10-diol, 1-dihydrogenophosphate 10- (6-aminohexanoate) (WO 01/46127).

Generally, 3D-MPL is used as the antigen, and 3D-MPL is delivered in an oil in water emulsion or a complex oil in water emulsion. Fusion of 3D-MPL is advantageous because it becomes a stimulator of T-cell response effectors.

Other TLR4 ligands that may be used are alkyl glucosamide phosphates (AGP), such as those disclosed in WO9850399 or US6303347 (also described in the process for preparing AGP), or pharmaceutically acceptable salts of AGP disclosed in US6764840. to be. Some AGPs are TLR4 agonists and some are TLR4 antagonists. Both are thought to be useful as an adjuvant.

Another immunostimulant for use in the present invention is Quil A and its derivatives. Quil A is a tree in South America, Quilaja Saponaria Molina ( Quilaja) Saponaria Saponin preparation isolated from Molina ), see Dalsgaard et al. in 1974 (“Saponin adjuvants”, Archiv fur die gesamte Virusforschung, Vol. 44, Springer Verlag, Berlin, p243-254), was first described as having adjuvant activity. Purified fragments of Quil A, such as QS7 and QS21 (also known as QA7 and QA21), that retain adjuvant activity without toxicity associated with Quil A, were isolated by HPLC (EP 0 362 278). QS-21 is a natural saponin derived from the bark of Quilaza saponaria Molina that induces CD8 + cytotoxic T cells (CTL), Th1 cells and predominant IgG2a antibody responses.

Certain formulations of QS21 have been described and these formulations further comprise sterols (WO 96/33739). The ratio of QS21: sterol will generally be from about 1: 100 to 1: 1 weight to weight. Generally excess sterol is present and the ratio of QS21: sterol is at least 1: 2 w / w. In general, for human administration, QS21 and sterol will be present in the vaccine in the range of about 1 μg to about 100 μg, such as about 10 μg to about 50 μg per dose.

Liposomal formulations generally include natural lipids such as phosphatidylcholine that is non-crystalline at room temperature, such as egg yolk phosphatidylcholine, dioleoyl phosphatidylcholine or dilauryl phosphatidylcholine. Liposomes can also include charged lipids that increase the stability of the liposome-QS21 structure for liposomes composed of saturated lipids. In this case, the amount of charged lipids is usually 1-20% w / w, such as 5-10%. The ratio of sterols to phospholipids is 1-50% (mol / mol), such as 20-25%.

This composition may comprise MPL (3-diacylated mono-phosphoryl lipid A, also known as 3D-MPL). 3D-MPL is known from GB 2 220 211 (Ribi), as a mixture of type 3 de-O-acylated monophosphoryl lipid A with 4, 5, or 6 acylated chains, by Ribi Immunochem, Montana Are manufactured.

Saponins can be in the form of micelles, mixed micelles (typically but not exclusively, comprising bile salts), ISCOM matrices (EP 0 109 942), liposomes or related colloidal structures, such as worm-like forms. ) Or ring-formed multimeric complexes or lipids and lipids / layered structures and lamellae, or oil-in-water emulsion forms (eg WO 95/17210). have.

Usually, saponins are present in the form of liposome formulations, ISCOMs or oil-in-water emulsions.

Immunostimulatory oligonucleotides may also be used. Examples of oligonucleotides for use in a vaccine or adjuvant of the invention include CpG containing oligonucleotides, which are generally two or more dinucleotide CpGs separated into three or more, more preferably six or more nucleotides Contains motifs. CpG motifs are cytosine nucleotides followed by guanine nucleotides. CpG oligonucleotides are generally deoxynucleotides. In a first embodiment, internucleotide phosphorothiothioate, or more preferably phosphorothioate, bonds in oligonucleotides, although phosphodiester and other internucleotide linkages are also within the scope of the present invention. Oligonucleotides within mixed internucleotide bonds are also included within the scope of the present invention. Processes for producing phosphorothioate oligonucleotides or phosphorodithioates are described in US 5,666,153, US 5,278,302 and WO 95/26204.

Examples of oligonucleotides are as follows:

The sequence may comprise phosphorothioate modified internucleotide bonds.

Alternative CpG oligonucleotides may comprise one or more of the above sequences and have minor deletions or additions to them.

CpG oligonucleotides can be synthesized by any method known in the art (see eg EP 468520). Conveniently, such oligonucleotides can be synthesized using an automated synthesizer.

Examples of TLR 2 agonists include peptidoglycans or lipoproteins.

Imidazoquinolines such as Imiquimod and Resiquimod are known as TLR7 agonists. Single-stranded RNA is also known as TLR agonist (TLR8 in humans and TLR7 in mice), while double-stranded RNA and poly ICs (imiles of polyinosonic-polycytidylic acid-commercially synthesized viral RNA) One example is a TLR 3 agonist. 3D-MPL is an example of a TLR4 agonist, while CpG is an example of a TLR9 agonist.

Immunostimulants may alternatively or additionally be included. In a first embodiment, this immune stimulant will be 3-diacylated monophosphoryl lipid A (3D-MPL).

In a first aspect, the adjuvant comprises 3D-MPL.

In a first aspect, the adjuvant comprises QS21.

In a first aspect, the adjuvant comprises CpG.

In a first embodiment, the adjuvant is formulated in an oil-in-water emulsion.

In a first embodiment, the adjuvant is formulated into liposomes.

Adjuvant combinations include 3D-MPL and QS21 (EP 0 671 948 Bl) oil-in-water emulsions or 3D-MPL (EP 0 689 454 Bl) formulated with liposome formulations or other carriers including 3D-MPL and QS21. do. Other preferred adjuvant systems include combinations of 3D-MPL, QS21 and CpG oligonucleotides as described in US 6558670 and US 6544518.

Formulation

Vaccine preparation is generally described in New Trends and Developments in Vaccines, edited by Voller et al., University Park Press, Baltimore, Maryland, U.S.A., 1978. Encapsulation into liposomes is described, eg, US Pat. No. 4,235,877 to Fullerton.

The formulations of the present invention can be used for both prophylactic and therapeutic purposes. The present invention therefore provides a vaccine composition as described herein for medical use, such as for the treatment and / or prevention of malaria.

In a first aspect, the invention comprises a C7 adenovirus vector according to the invention and a malaria antigen such as a novel antigen of RTS, S or Seq ID No: 1 or the same virus like particles and excipients, optionally an adjuvant It provides a composition.

Immunogenic in this specification is intended to refer to the ability to induce an immune response, which response is specific to the malaria component in the appropriate formulation. This reaction may require the presence of appropriate adjuvant and / or boosting. Boosters, such as those containing doses that are less or similar to conventional doses, may be required to obtain an appropriate immunogenic response.

Compositions / pharmaceutical formulations according to the invention may also comprise one or more additional antigens in the mixture, such as antigens derived from P. falciparum and / or P. vivax , for example such antigens. Is selected from DBP, PvTRAP, PvMSP2, PvMSP4, PvMSP5, PvMSPβ, PvMSP7, PvMSP8, PvMSP9, PvAMAl and RBP or fragments thereof.

In another example, antigens derived from P. falciparum include PfEMP-1, Pfs 16 antigen, MSP-1, MSP-3, LSA-1, LSA-3, AMA-1 and TRAP. Other Plasmodium antigens are P. falciparum EBA, GLURP, RAP1, RAP2, Sequencerin, Pf332, STARP, SALSA, PfEXP1, Pfs25, Pfs28, PFS27 / 25, Pfs48 / 45, Pfs230 and other Plasma Their analogues in the medium spp.

The invention also relates to the use of C7 for encoding a malaria antigen, for example for the treatment and / or prophylaxis of malaria, especially as described herein, or for the manufacture of a medicament of the same use.

The invention also includes a method of treatment comprising administering a therapeutically effective amount of one or more aspects of the invention. Optionally, the C7 virus vector according to the present invention may be a malaria antigen such as RTS, S or Seq ID No. The antigen of 1 can be co-administered or co-formulated, optionally in the presence of an adjuvant comprising eg 3D-MPL and / or savonin such as QS21.

C7 vectors can also be co-administered or co-formulated with another adenovirus vector, which encodes the same different antigen of different serotypes and / or origins.

The invention also extends to the use of any embodiment as defined herein in a prime boost regime, such that the priming dose or dosages are given at time point 0 (and subsequent primes are, for example, within 3 months). The boost is administered, for example, at about 4, 5, 6, 7, 8, 9, 10, 11 or 12 weeks after the last priming dose, optionally with additional boosting shots or shots up to after the initial boosting shot. It is administered after one year.

Advantageously, one or more embodiments of the present invention comprising the combination vaccines described above are directed to specific body fluids (ie, antibody responses) and / or cellular immune responses (eg, CD8 + and / or CD4 +), such as antibody responses and CD 8+ and And / or stimulate the CD4 + response, in particular the CD8 + and antibody responses. In other words, the response is specific to the CS protein and / or S antigen (as appropriate).

This type of balanced immune response may be required to provide so-called aseptic protection against malaria infection.

In addition, the antibody response to the combination may be enhanced with respect to the antibody response to the regimen of the adjuvanted protein only.

In a first embodiment, the present invention provides the use of C7 as a prime or boost in a prime boost regimen:

C7 adenovirus vectors encoding the same or different malaria antigens,

Simian of another viral vector, such as a human adenovirus vector, such as Ad5 or Ad35, that encodes a CS protein from a malaria antigen, such as P. falciparum , or a different serotype (ie, not C7) Adenovirus vectors and / or

Includes malaria antigens such as RTS, S and adjuvants, saponins and / or 3D-MPL,

As a complementary component of the regimen.

The invention also provides any of the embodiments described above for the manufacture of a medicament for the treatment and / or prevention of malaria infection.

dose

The amount of 3D-MPL used is generally small but depending on the vaccine formulation can be in the range of 1-1000 μg per dose, such as 1-500 μg per dose, and for example in the range of 1-100 μg per dose, such as 50 or 25 μg per dose. have.

The amount of CpG or immunostimulatory oligonucleotides in the adjuvant or vaccine of the invention is generally small, but depending on the vaccine formulation, ranges from 1-1000 μg per dose, such as 1-500 μg per dose, and eg 1-100 μg per dose. It can be a range.

The amount of saponin for use in the adjuvant of the invention ranges from 1-1000 μg per dose, such as 1-500 μg per dose, such as 1-250 μg per dose, and especially 1-100 μg per dose, such as 50 per dose. Or 25 μg.

When the protein is administered, the dosage is, for example, 1 to 500 μg, such as 10 to 100 μg, in particular 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75 or 80 μg per dose. Can be.

When an adenovirus vector is administered, the dosage may be, for example, 10 ³ to 10 ¹⁶ vpu, such as 10 ⁶ to 10 ¹⁰ vpu.

When used in combination, the amount used for each component of the combination will correspond to the dosage given to the component alone.

The invention also extends to kits comprising the factors used in the combination according to the invention.

The present invention relates to a process for the preparation of adenovirus vectors according to the invention and to formulations comprising adenovirus vectors.

The invention also relates to a method for producing a protein of Seq ID No 1.

In the text of this specification, "comprising" is interpreted as "including".

The invention encompasses certain factors, but extends to embodiments corresponding to the embodiments described herein consisting or consisting substantially of the relevant factors.

The discussion in the Background section of this specification has been provided for the purpose of incorporating the invention into the text. It is not a confession of the prior art, but especially of constructing common knowledge.

The following examples are presented to illustrate the methodology that can be used herein.

Example

Example 1

The synthetic gene was manufactured by Medigenomix. The gene was cloned into pCR2.1 -TOPO-TA cloning vector (see Invitrogen Figure 1). This vector was cleaved with Not I and Bam HI and a recombinant shuttle plasmid vector (-Ade2) was constructed. A map of the shuttle plasmid is shown in FIG.

Expression cassette and virus extraction ( rescue Description of methodology for

Expression cassettes include cytomegalovirus (CMV) early promoter and first exon, introns derived from plasmid pCI (purchased from Promega), DNA encoding Ade2, and rabbit globin polyadenylation signals. The complete cassette is flanking the recognition site for each of the restriction enzymes I-CeuI and PI-SceI. The expression cassette was removed from the shuttle plasmid using I-CeuI and PI-SceI and described by Roy et al. Hum Gene Ther. (2004) 5: 519-530, incorporated into the plasmid molecular clone of the E1 deleted genome of SAdV-24 (ie C7) -pC7 000 pkGFP to obtain the plasmid shown in FIG. 3. The plasmid molecular clone DNA was cut and linearized with restriction enzyme PacI and transfected with HEK 293 cells to extract recombinant adenovirus. The adenovirus was propagated, amplified and purified using standard techniques.

The sequence of the expression cassette for the recognition site from I-CeuI to PI-Sce is shown in Seq ID Nos: 18.

Example 2

C57B1 / 6 in mouse C7 - Ade1  And C7 - Of Ade2  Immunogenicity

C57B1 / 6 mice were inoculated intramuscularly at a dose range (10e10, 10e9, 10e8 virus particles) of C7 chimpanzee adenovirus expressing both constructs Ade1 or Ade2. As a positive control, several mice were inoculated with human adenovirus 5 (dose 10e9 and 10e8) expressing both Ade1 or Ade2. As a negative control, some mice were inoculated with C7 deficient and Ad5 deficient viral vectors.

Peripheral blood was collected and grouped 14, 28, 34 and 49 days post inoculation, and Ag-specific CD4 and CD8 T cell responses producing IL-2 and / or IFN-gamma were of interest in vitro overnight After restimulation with a pool of 15mer peptides comprising the sequence, i.e., the N-terminal region (N-terminus) or C-terminal region (C-terminus) of the CS protein, Measured by an analyzer. As a negative control, some cells were also cultured overnight in vitro in culture medium (unstimulated). Ag-specific responses were calculated by subtracting the average cytokine response produced by unstimulated cells from the average cytokine response produced by peptide-stimulated cells.

The results show that under the experimental conditions described above, the two constructs induce CS-specific CD4 and CD8 T cell responses (FIGS. 4-15). Among them, only C-terminal specific CD8 T cell responses were detected in mice inoculated with adenovirus carrying Ade2 inserts (FIGS. 4 and 8). In particular, this C-terminal specific CD8 T cell response was similar in mice inoculated with lOelOvp of C7 Ade2 or 10e9vp of Ad5 Ade2 (FIG. 8).

In addition, anti-CS antibody responses were measured by ELISA in serum collected 48 days after inoculation. In particular, the total Ig response to the R32LR polypeptide (including the middle portion of the P. falciparum CSP) was measured (Mettens et al., Vaccine 2008). This result indicates that single inoculation with C7 Adel or C7 Ade2 induces low levels of R32LR-specific antibody responses. The intensity of this reaction correlates with the number of virus particles used for inoculation (dose range effect).

Example 3

CB6F1  From the mouse C7 - Ade2 Immunogenicity of

CB6F1 mice were inoculated intramuscularly at a dose range of C7 chimpanzee adenovirus expressing Ade2 constructs (10e10, 10e9, 10e8 virus particles) (5 pools of mice / group). Peripheral blood was collected and grouped 21, 28 and 35 days post inoculation, and CS C-terminal and CS N-terminal-specific CD4 and CD8 T cell responses producing IL-2 and / or IFN-gamma Was restimulated in vitro overnight with a pool of 15mer peptides containing the sequence of interest, namely the N-terminal region (N-terminus) or C-terminal region (C-terminus) of the CS protein. It was then measured by flow cytometry. As a negative control, some cells were also cultured overnight in vitro in culture medium (unstimulated). Ag-specific responses were calculated by subtracting the average cytokine response produced by unstimulated cells from the average cytokine response produced by peptide-stimulated cells.

The results indicate that in this mouse lineage, a single inoculation of 10e9vp or lOelOvp of C7 Ade2 induces C-terminal and N-terminal specific CD4 and CD8 T cell responses (FIGS. 17-20). In particular, the average intensity of the observed response is the same, if not high, when the 10e9 vp dose is used. CS-specific CD8 T cell responses are mainly N-terminal specific, while CS-specific CD4 T cell responses likewise target the N-terminal and C-terminal regimens of CS proteins.

Cytokine profiles of CS-specific CD4 and CD8 T cell responses were also measured, which were similar throughout all time points tested. The profiles shown after 28 days of inoculation are shown in FIGS. 21-24, which represent other tested time points. Briefly, CS-specific CD8 T cell responses consist mostly of CD8 T cells producing only IFNg (FIGS. 21 and 22). In addition, the CS-specific CD4 T cell response consisted of IFNg producing CD4 T cells as well as smaller amounts of CD4 T cells producing IL2 alone or both IL2 and IFNg (FIGS. 23 and 24).

Example 4

Prime Of Boost  of mine C7 - Ade2 Immunogenicity of or CB6F1  Within mouse RTS , S / AS01B With concurrent- Formulation

We tested the immunogenicity of C7 chimpanzee adenoviruses expressing Ade2 construct co-formulated (combo) with RTS, S / AS01B in prime-boost or CB6F1 mice. ASOlB is an adjuvant system comprising 3D-MPL and QS21 formulated with liposomes. Mice were inoculated intramuscularly at days 0, 14 and 28 as follows:

In the table above,

A = 10e9 vp of C7 Ade2

P = 5 μg RTS, S / 50 μl AS01B

C = lOelO vp + 5 μg RTS, S / 50 μl ASOlB of C7 Ade2.

Peripheral blood was collected and grouped on days 21 (7d pII), 35 (7d pIII), 49 (21d pIII), 63 (35d pIII), 77 (49d pIII) and inoculated with IL-2 and / or IFN-gamma CS C-terminus, CS N-terminus, and HBs specific CD4 and CD8 T cell responses to produce a overnight in vitro into a pool of 15mer peptides containing the sequence of interest (CS N-terminus, CS C-terminus or HBs). After restimulation, it was measured by flow cytometry. As a negative control, some cells were also cultured overnight in vitro in culture medium (unstimulated). Ag-specific responses were calculated by subtracting the average cytokine response produced by unstimulated cells from the average cytokine response produced by peptide-stimulated cells.

The reaction shows the following:

APP, PPA and CCC all induce N-terminal specific CD8 T cell responses. At time point 7dpII, N-terminal specific CD8 T cell responses are observed in the CCC group followed by the APP group. At later time points tested (21d pIII, 35d pIII and 49d pIII), the reaction is present and remains of similar intensity in the APP, PPA and CCC groups (FIG. 25).

APP, PPA and CCC induce a C-terminal specific CD8 T cell response, which still persists 49 days after the third inoculation (FIG. 26).

N-term specific CD4 T cell responses are detected mainly in mice inoculated with APP or CCC, in the APP group this response is detected with higher intensity (FIG. 27).

All groups show C-terminal specific CD4 T cell responses. However, APP and CCC induce similar high levels of C-terminal specific CD4 T cell responses, which are about two to three times higher than responses induced by PPA and PPP at all time points tested.

HBs-specific CD4 and CD8 T cell responses are higher in animals inoculated with PPP than animals inoculated with APP, PPA or CCC (FIGS. 29 and 30).

CCC treatment regimens are the only ones involved in the simultaneous induction of CS and HBs-specific CD4 and CD8 T cell responses.

Cytokine profiles of CS- and HBs-specific CD4 and CD8 T cell responses were also measured, which were similar over the time points tested. Measurements from the 21 d pIII time point are shown below as representative of all time points tested (FIGS. 31-36). Briefly, Ag-specific CD8 T cell responses mostly consisted of IFNg producing CD8 T cells (FIGS. 31-33). Ag-specific CD4 T cell responses, on the other hand, consist of a mixture of IFNg, IFNg and IL-2 producing CD4 T cells, and contain less amount of IL-2 producing CD4 T cells (FIGS. 34-36). ).

In addition, Ag-specific antibody responses were measured by ELISA in serum collected 14 and 42 days after the third inoculation. In particular, the total Ig response to R32LR polypeptides (ie, including the middle portion of the P. falciparum CSP) and the total Ig response to HBs were measured (Mettens et al., Vaccine 2008). All immunization regimes induced R32LR and HBs-specific antibody responses that persisted up to the last tested time point (FIGS. 37 and 38).

Example 5

CB6F1  Simultaneous in mouse Formulation C7 - Ade2 + RTS , S / AS01B Immunogenicity of

In this experiment, we compared the immunogenicity of the co-formulated (combo) chimpanzee adenovirus C7 Ade2 with RTS, S / AS01B. In particular, we compared the immune response induced by 1, 2 or 3 injection of the combo in CB6F1 mice (4 pools of mice / group). In addition, they were evaluated at different intervals between two injections of the combo (ie, 14 days and 21 days). Finally, a group of mice inoculated with A-P-P served as a control in the experiment. The planned experiment can be summarized as follows:

In the table above,

A = 10e9 vp of C7 Ade2

P = 5 μg RTS, S / 50 μl AS01B

C = 10e9 vp + 5μg RTS of C7 Ade2, S / 50μl ASOlB

Peripheral blood was collected and grouped at 35, 42, 49, 63, and 98 days and CS C-terminal, CS N-terminal, and HBs specific CD4 and CD8 T cell responses producing IL-2 and / or IFN-gamma Was re-stimulated in vitro overnight with a pool of 15mer peptides containing the sequence of interest, namely the N-terminal region (N-terminus), C-terminal region (C-terminus) or HBs of the CS protein, followed by flow cytometry It was. As a negative control, some cells were also cultured overnight in vitro in culture medium (unstimulated). Ag-specific responses were calculated by subtracting the average cytokine response produced by unstimulated cells from the average cytokine response produced by peptide-stimulated cells.

The results indicate that under these experimental conditions, two immunizations of the combo are required to simultaneously induce CS C-terminus, CS N-terminus and HBs-specific CD4 and CD8 T cell responses. The interval between 2 immunizations of the combos did not appear to significantly affect the level of sensed Ag-specific T cell response. Mice inoculated three times with the combo tended to have higher HBs-specific CD4 and CD8 T cell responses. Kinetics of Ag-specific T cell responses are shown in FIGS. 39-44.

Cytokine profiles of CS- and HBs-specific CD4 and CD8 T cell responses were also measured, which were similar over the time points tested. Results from day 42 of the study are shown below as representative of all time points tested (FIGS. 42-47). Briefly, Ag-specific CD8 T cell responses mostly consisted of IFNg producing CD8 T cells (FIGS. 45-47). Ag-specific CD4 T cell responses, on the other hand, consisted of a mixture of CD4 T cells producing IFNg, IFNg and IL-2, and containing less amount of CD4 T cells producing IL-2 (FIGS. 48-48). 50).

In addition, Ag-specific antibody responses were measured by ELISA in serum collected on days 56 and 99 of the study. In particular, the total Ig response to R32LR polypeptides (ie, including the middle portion of the P. falciparum CSP) and the total Ig response to HBs were measured (Mettens et al., Vaccine 2008). All immunization regimes elicited R32LR and HBs-specific antibody responses: in each group this response showed similar intensity at these two time points tested. In addition, when comparing the groups inoculated with the combo, the response tended to be higher in the group inoculated with three doses of the combo (FIGS. 51 and 52).

Example 6

Combo  Each component, CB6F1  Above two in mouse CS  & HBs  T cells and Ab  To induce a response C7 - Ade2 , RTS9S  And ASOlB The need for

In this experiment, we assessed the need for each component of the combo (ie C7 Ade2, RTS, S and ASOlB) to induce CS and HBs-specific CD4 and CD8 T cell responses simultaneously. In this experiment, CB6F1 mice (6 pools of mice / group) were inoculated twice intramuscularly (day 0 and day 14) with combos or components thereof as shown below.

In the table above,

A = 10e9 vp of C7 Ade2

P = 5 μg RTS, S / 50 μl AS01B

C = 10e9 vp + 5μg RTS of C7 Ade2, S / 50μl ASOlB

C7 deficiency = 10e9 vp (no insertion) of C7 deficiency vector

RTS, S = 5μg RTS, S

ASOlB = GSK Proprietary Adjuvant System 1B

Buffer = ASOlB Buffer (without immune stimulant)

Peripheral blood was collected and grouped on days 14, 28, 70, 91 and 112, and the CS C-terminal, CS N-terminal, and HBs specific CD4 and CD8 T cell responses producing IL-2 and / or IFN-gamma Was re-stimulated in vitro overnight with a pool of 15mer peptides containing the sequence of interest, namely the N-terminal region (N-terminus), C-terminal region (C-terminus) or HBs of the CS protein, followed by flow cytometry It was. As a negative control, some cells were also cultured overnight in vitro in culture medium (unstimulated). Ag-specific responses were calculated by subtracting the average cytokine response produced by unstimulated cells from the average cytokine response produced by peptide-stimulated cells.

In addition, Ag-specific antibody responses were measured by ELISA in serum collected on days 42 and 84 of the study. In particular, the total Ig response to R32LR polypeptides (ie, including the middle portion of the P. falciparum CSP) and the total Ig response to HBs were measured (Mettens et al., Vaccine 2008).

The results indicate that each component of the combo is required to induce R32LR and HBs antibody responses (FIGS. 65 and 66) simultaneously, including CS (N-terminal and C-terminal) CD4 and CD8 T cell responses (FIGS. 53-58). Indicates.

Average cytokine profiles of CS- and HBs-specific CD4 and CD8 T cell responses were also measured at each time point in the study, which is shown in FIGS. 59-64. Briefly, Ag-specific CD8 T cell responses mostly consisted of IFNg producing CD8 T cells (FIGS. 59-61). Ag-specific CD4 T cell responses, on the other hand, consist of a mixture of CD4 T cells producing IFNg, IFNg and IL-2, and contain less amount of CD4 T cells producing IL-2 (FIGS. 62-62). 64).

Example 7

CB6F1  Synthetic in mouse C7 Ade2 Immunogenicity of

Synthetic C7 chimpanzee adenoviruses expressing Ade2 constructs could be constructed and their immunogenicity compared to that of native C7 Ade2. In this experiment, CB6F1 mice (6 pools of mice / group) were inoculated with 10e9 vp of native C7 Ade2 or synthetic mimetics thereof.

Peripheral blood was collected and grouped 21, 28 and 35 days post inoculation, and CS C-terminal, CS N-terminal CD4 and CD8 T cell responses producing IL-2 and / or IFN-gamma were of interest in vitro overnight. The sequence was restimulated with a pool of 15mer peptides comprising the N-terminal region (N-terminus) and C-terminal region (C-terminus) of the CS protein, followed by flow cytometry. As a negative control, some cells were also cultured overnight in vitro in culture medium (unstimulated). Ag-specific responses were calculated by subtracting the average cytokine response produced by unstimulated cells from the average cytokine response produced by peptide-stimulated cells.

The results indicate that the two C7 adenoviruses induced similar levels of N-terminal and C-terminal specific CD4 and CD8 T cell responses. In particular, despite the viral vectors used (raw or synthetic), Ag-specific CD8 T cell responses were predominantly N-terminal specific (FIGS. 67 and 68). N-terminal and C-terminal specific CD4 T cell responses showed similar intensity regardless of the viral vector used (FIGS. 69 and 70).

Cytokine profiles of CS-specific CD4 and CD8 T cell responses were measured and shown in FIGS. 71-74. Briefly, Ag-specific CD8 T cell responses mostly consisted of CD8 T cells producing IFNg (FIGS. 71 and 72). Ag-specific CD4 T cell responses, on the other hand, Ag-specific CD4 T cell responses consisted of a mixture of CD4 T cells producing IFNg, IFNg and IL-2, and more IL-2 producing CD4 T cells. In small amounts (FIGS. 73-74).

SEQUENCE LISTING <110> Cohen, Joseph D.            Marchand, Martine   <120> Vaccine <130> VB62729 <150> US60 / 992802 <151> 2007-12-06 <160> 19 <170> PatentIn version 3.5 <210> 1 <211> 382 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Ade2 protein <400> 1 Met Met Arg Lys Leu Ala Ile Leu Ser Val Ser Ser Phe Leu Phe Val 1 5 10 15 Glu Ala Leu Phe Gln Glu Tyr Gln Cys Tyr Gly Ser Ser Ser Asn Thr             20 25 30 Arg Val Leu Asn Glu Leu Asn Tyr Asp Asn Ala Gly Thr Asn Leu Tyr         35 40 45 Asn Glu Leu Glu Met Asn Tyr Tyr Gly Lys Gln Glu Asn Trp Tyr Ser     50 55 60 Leu Lys Lys Asn Ser Arg Ser Leu Gly Glu Asn Asp Asp Gly Asn Asn 65 70 75 80 Asn Asn Gly Asp Asn Gly Arg Glu Gly Lys Asp Glu Asp Lys Arg Asp                 85 90 95 Gly Asn Asn Glu Asp Asn Glu Lys Leu Arg Lys Pro Lys His Lys Lys             100 105 110 Leu Lys Gln Pro Ala Asp Gly Asn Pro Asp Pro Asn Ala Asn Pro Asn         115 120 125 Val Asp Pro Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn Pro Asn     130 135 140 Val Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 145 150 155 160 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn                 165 170 175 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn             180 185 190 Val Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn         195 200 205 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn     210 215 220 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 225 230 235 240 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn                 245 250 255 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Lys Asn Asn Gln             260 265 270 Gly Asn Gly Gln Gly His Asn Met Pro Asn Asp Pro Asn Arg Asn Val         275 280 285 Asp Glu Asn Ala Asn Ala Asn Ser Ala Val Lys Asn Asn Asn Asn Glu     290 295 300 Glu Pro Ser Asp Lys His Ile Lys Glu Tyr Leu Asn Lys Ile Gln Asn 305 310 315 320 Ser Leu Ser Thr Glu Trp Ser Pro Cys Ser Val Thr Cys Gly Asn Gly                 325 330 335 Ile Gln Val Arg Ile Lys Pro Gly Ser Ala Asn Lys Pro Lys Asp Glu             340 345 350 Leu Asp Tyr Ala Asn Asp Ile Glu Lys Lys Ile Cys Lys Met Glu Lys         355 360 365 Cys Ser Ser Val Phe Asn Val Val Asn Ser Ala Ile Gly Leu     370 375 380 <210> 2 <211> 1149 <212> DNA <213> Artificial Sequence <220> <223> Ade2 nucleic acid sequence <400> 2 atgatgagaa aacttgccat cctcagcgtc agctctttcc tgttcgtgga ggccctcttc 60 caggagtatc agtgctacgg aagcagcagc aatacaaggg tcctgaacga gctcaactat 120 gacaacgctg gaacgaacct gtataacgag ctggagatga actactatgg caagcaggag 180 aactggtata gcctgaagaa gaacagccgg tccctgggcg agaacgacga cggcaacaac 240 aacaacggcg acaacggcag ggagggcaaa gatgaggaca agagggacgg gaacaacgag 300 gataacgaga agctgcggaa gcccaagcac aagaaactca agcagcccgc cgacgggaac 360 ccggacccca atgcaaatcc caacgtcgac ccaaacgcaa accctaacgt ggaccccaac 420 gccaatccca acgtcgatcc taatgccaat ccaaatgcca accctaacgc aaatcctaat 480 gcaaacccca acgccaatcc taacgccaac ccaaatgcca acccaaacgc taaccccaac 540 gctaacccaa atgcaaatcc caatgctaac ccaaacgtgg accctaacgc taaccccaac 600 gcaaacccta acgccaatcc taacgcaaac cccaatgcaa acccaaacgc aaatcccaac 660 gctaacccta acgcaaaccc caacgccaac cctaatgcca accccaatgc taaccccaac 720 gccaatccaa acgcaaatcc aaacgccaac ccaaatgcaa accccaacgc taatcccaac 780 gccaacccaa acgccaatcc taacaagaac aatcagggca acgggcaggg ccataacatg 840 ccgaacgacc ctaatcggaa tgtggacgag aacgccaacg ccaacagcgc cgtgaagaac 900 aacaacaacg aggagccctc cgacaagcac atcaaggaat acctgaacaa gatccagaac 960 agtctgagca ccgagtggtc cccctgctcc gtgacctgcg gcaacggcat ccaggtgagg 1020 atcaagcccg gctccgccaa caagcccaag gacgagctgg actacgccaa cgacatcgag 1080 aagaagatct gcaagatgga gaaatgcagc tctgtgttca acgtcgtgaa ctccgccatc 1140 ggcctgtga 1149 <210> 3 <211> 372 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Ade1 protein <400> 3 Met Met Arg Lys Leu Ala Ile Leu Ser Val Ser Ser Phe Leu Phe Val 1 5 10 15 Glu Ala Leu Phe Gln Glu Tyr Gln Cys Tyr Gly Ser Ser Ser Asn Thr             20 25 30 Arg Val Leu Asn Glu Leu Asn Tyr Asp Asn Ala Gly Thr Asn Leu Tyr         35 40 45 Asn Glu Leu Glu Met Asn Tyr Tyr Gly Lys Gln Glu Asn Trp Tyr Ser     50 55 60 Leu Lys Lys Asn Ser Arg Ser Leu Gly Glu Asn Asp Asp Gly Asn Asn 65 70 75 80 Asn Asn Gly Asp Asn Gly Arg Glu Gly Lys Asp Glu Asp Lys Arg Asp                 85 90 95 Gly Asn Asn Glu Asp Asn Glu Lys Leu Arg Lys Pro Lys His Lys Lys             100 105 110 Leu Lys Gln Pro Ala Asp Gly Asn Pro Asp Pro Asn Ala Asn Pro Asn         115 120 125 Val Asp Pro Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn Pro Asn     130 135 140 Val Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 145 150 155 160 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn                 165 170 175 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn             180 185 190 Val Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn         195 200 205 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn     210 215 220 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn 225 230 235 240 Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn                 245 250 255 Ala Asn Pro Asn Ala Asn Lys Asn Asn Gln Gly Asn Gly Gln Gly His             260 265 270 Asn Met Pro Asn Asp Pro Asn Arg Asn Val Asp Glu Asn Ala Asn Ala         275 280 285 Asn Ser Ala Val Lys Asn Asn Asn Asn Glu Glu Pro Ser Asp Lys His     290 295 300 Ile Lys Glu Tyr Leu Asn Lys Ile Gln Asn Ser Leu Ser Thr Glu Trp 305 310 315 320 Ser Pro Cys Ser Val Thr Cys Gly Asn Gly Ile Gln Val Arg Ile Lys                 325 330 335 Pro Gly Ser Ala Asn Lys Pro Lys Asp Glu Leu Asp Tyr Ala Asn Asp             340 345 350 Ile Glu Lys Lys Ile Cys Lys Met Glu Lys Cys Ser Ser Val Phe Asn         355 360 365 Val Val Asn Ser     370 <210> 4 <211> 1119 <212> DNA <213> Artificial Sequence <220> <223> Ade1 nucleic acid sequence <400> 4 atgatgagga aactggccat cctgagcgtg agcagcttcc tgttcgtgga ggccctgttt 60 caggagtacc agtgctacgg cagcagcagc aacacccggg tgctgaacga gctgaactac 120 gacaacgccg gcaccaacct gtacaacgag ctggagatga actactacgg caagcaggag 180 aactggtaca gcctgaagaa gaacagccgg tctctgggcg agaacgacga cggcaacaac 240 aacaacggcg acaacggccg ggagggcaag gacgaggaca agcgggacgg caacaacgag 300 gacaacgaga agctgcggaa gcccaagcac aagaaactta agcagcccgc cgacggcaac 360 cccgacccca acgccaaccc caacgtggac cccaacgcca atcctaatgt cgaccccaat 420 gccaatccga acgttgatcc caatgcgaat cctaacgcta accccaatgc caacccaaat 480 gccaatccaa atgcaaatcc caacgccaat ccaaacgcaa accctaatgc taatccaaac 540 gctaatccta atgccaatcc caatgctaac ccaaacgtcg atcctaacgc aaatccgaac 600 gctaacccca acgcaaatcc caacgctaac ccgaacgcaa accctaacgc caatccgaat 660 gccaacccaa acgccaaccc gaacgctaat ccgaatgcta acccgaatgc taatcctaac 720 gcaaacccaa atgcaaaccc caatgcaaac ccgaacgcca atcccaacgc caatcctaat 780 gccaacaaga acaatcaggg caacggccag ggccacaaca tgcccaacga ccccaaccgg 840 aacgtggacg agaacgccaa cgccaacagc gccgtgaaga acaacaacaa cgaggagccc 900 agcgacaagc acatcaagga gtacctgaac aagatccaga acagcctgag caccgagtgg 960 agcccctgca gcgtgacctg cggcaacggc attcaggtgc ggatcaagcc cggcagcgcc 1020 aacaagccca aggacgagct ggactacgcc aatgacatcg agaagaagat ctgcaagatg 1080 gagaagtgca gcagcgtgtt caacgtggtg aactcctga 1119 <210> 5 <211> 314 <212> PRT <213> chimpanzee adenovirus Pan7 <400> 5 Ala Pro Lys Gly Ala Pro Asn Thr Cys Gln Trp Thr Tyr Lys Ala Gly 1 5 10 15 Asp Thr Asp Thr Glu Lys Thr Tyr Thr Tyr Gly Asn Ala Pro Val Gln             20 25 30 Gly Ile Ser Ile Thr Lys Asp Gly Ile Gln Leu Gly Thr Asp Ser Asp         35 40 45 Gly Gln Ala Ile Tyr Ala Asp Glu Thr Tyr Gln Pro Glu Pro Gln Val     50 55 60 Gly Asp Ala Glu Trp His Asp Ile Thr Gly Thr Asp Glu Lys Tyr Gly 65 70 75 80 Gly Arg Ala Leu Lys Pro Asp Thr Lys Met Lys Pro Cys Tyr Gly Ser                 85 90 95 Phe Ala Lys Pro Thr Asn Lys Glu Gly Gly Gln Ala Asn Val Lys Thr             100 105 110 Glu Thr Gly Gly Thr Lys Glu Tyr Asp Ile Asp Met Ala Phe Phe Asp         115 120 125 Asn Arg Ser Ala Ala Ala Ala Gly Leu Ala Pro Glu Ile Val Leu Tyr     130 135 140 Thr Glu Asn Val Asp Leu Glu Thr Pro Asp Thr His Ile Val Tyr Lys 145 150 155 160 Ala Gly Thr Asp Asp Ser Ser Ser Ser Ile Asn Leu Gly Gln Gln Ser                 165 170 175 Met Pro Asn Arg Pro Asn Tyr Ile Gly Phe Arg Asp Asn Phe Ile Gly             180 185 190 Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly Val Leu Ala Gly Gln         195 200 205 Ala Ser Gln Leu Asn Ala Val Val Asp Leu Gln Asp Arg Asn Thr Glu     210 215 220 Leu Ser Tyr Gln Leu Leu Leu Asp Ser Leu Gly Asp Arg Thr Arg Tyr 225 230 235 240 Phe Ser Met Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro Asp Val Arg                 245 250 255 Ile Ile Glu Asn His Gly Val Glu Asp Glu Leu Pro Asn Tyr Cys Phe             260 265 270 Pro Leu Asp Ala Val Gly Arg Thr Asp Thr Tyr Gln Gly Ile Lys Ala         275 280 285 Asn Gly Asp Asn Gln Thr Thr Trp Thr Lys Asp Asp Thr Val Asn Asp     290 295 300 Ala Asn Glu Leu Gly Lys Gly Asn Pro Phe 305 310 <210> 6 <211> 179 <212> PRT <213> chimpanzee adenovirus Pan7 <400> 6 Thr Leu Trp Thr Thr Ala Asp Pro Ser Pro Asn Cys Lys Ile Tyr Ser 1 5 10 15 Glu Lys Asp Ala Lys Leu Thr Leu Cys Leu Thr Lys Cys Gly Ser Gln             20 25 30 Ile Leu Gly Thr Val Thr Val Leu Ala Val Asn Asn Gly Ser Leu Asn         35 40 45 Pro Ile Thr Asn Thr Val Ser Thr Ala Leu Val Ser Leu Lys Phe Asp     50 55 60 Ala Ser Gly Val Leu Leu Ser Ser Ser Le Le Asp Lys Glu Tyr Trp 65 70 75 80 Asn Phe Arg Lys Gly Asp Val Thr Pro Ala Glu Pro Tyr Thr Asn Ala                 85 90 95 Ile Gly Phe Met Pro Asn Ile Lys Ala Tyr Pro Lys Asn Thr Ser Ala             100 105 110 Ala Ser Lys Ser His Ile Val Ser Gln Val Tyr Leu Asn Gly Asp Glu         115 120 125 Ala Lys Pro Leu Met Leu Ile Ile Thr Phe Asn Glu Thr Glu Asp Ala     130 135 140 Thr Cys Thr Tyr Ser Ile Thr Phe Gln Trp Lys Trp Asp Ser Thr Lys 145 150 155 160 Tyr Thr Gly Glu Thr Leu Ala Thr Ser Ser Phe Thr Phe Ser Tyr Ile                 165 170 175 Ala Gln Glu              <210> 7 <211> 19 <212> PRT <213> Plasmodium falciparum <400> 7 Asn Asn Gly Asp Asn Gly Arg Glu Gly Lys Asp Glu Asp Lys Arg Asp 1 5 10 15 Gly asn asn              <210> 8 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence derived from P. falciparum CS protein <400> 8 Ala Ile Gly Leu One <210> 9 <211> 122 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence derived from P. falciparum CS protein <400> 9 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn 1 5 10 15 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn             20 25 30 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Val Asp         35 40 45 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn     50 55 60 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn 65 70 75 80 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn                 85 90 95 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn             100 105 110 Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn         115 120 <210> 10 <211> 144 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence derived from P. falciparum CS protein <400> 10 Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn Pro Asn Val Asp Pro 1 5 10 15 Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro             20 25 30 Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro         35 40 45 Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro     50 55 60 Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro 65 70 75 80 Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro                 85 90 95 Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro             100 105 110 Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro         115 120 125 Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro     130 135 140 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CpG oligonucleotide <400> 11 tccatgacgt tcctgacgtt 20 <210> 12 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> CpG oligonucleotide <400> 12 tctcccagcg tgcgccat 18 <210> 13 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> CpG oligonucleotide <400> 13 accgatgacg tcgccggtga cggcaccacg 30 <210> 14 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> CpG oligonucleotide <400> 14 tcgtcgtttt gtcgttttgt cgtt 24 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CpG oligonucleotide <400> 15 tccatgacgt tcctgatgct 20 <210> 16 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> CpG oligonucleotide <400> 16 tcgacgtttt cggcgcgcgc cg 22 <210> 17 <211> 2250 <212> DNA <213> Artificial Sequence <220> <223> Ade 1 expression cassette <400> 17 taactataac ggtcctaagg tagcgaaagc tcagatcggc tgaccgccca acgacccccg 60 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 120 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 180 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 240 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc 300 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc 360 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 420 tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag 480 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg 540 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg 600 cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgcgg ccagctttat 660 tgcggtagtt tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc 720 gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag 780 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt 840 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca 900 ctcccagttc aattacagct cttaaggcta gagtggccgc accatgatga ggaaactggc 960 catcctgagc gtgagcagct tcctgttcgt ggaggccctg tttcaggagt accagtgcta 1020 cggcagcagc agcaacaccc gggtgctgaa cgagctgaac tacgacaacg ccggcaccaa 1080 cctgtacaac gagctggaga tgaactacta cggcaagcag gagaactggt acagcctgaa 1140 gaagaacagc cggtctctgg gcgagaacga cgacggcaac aacaacaacg gcgacaacgg 1200 ccgggagggc aaggacgagg acaagcggga cggcaacaac gaggacaacg agaagctgcg 1260 gaagcccaag cacaagaaac ttaagcagcc cgccgacggc aaccccgacc ccaacgccaa 1320 ccccaacgtg gaccccaacg ccaatcctaa tgtcgacccc aatgccaatc cgaacgttga 1380 tcccaatgcg aatcctaacg ctaaccccaa tgccaaccca aatgccaatc caaatgcaaa 1440 tcccaacgcc aatccaaacg caaaccctaa tgctaatcca aacgctaatc ctaatgccaa 1500 tcccaatgct aacccaaacg tcgatcctaa cgcaaatccg aacgctaacc ccaacgcaaa 1560 tcccaacgct aacccgaacg caaaccctaa cgccaatccg aatgccaacc caaacgccaa 1620 cccgaacgct aatccgaatg ctaacccgaa tgctaatcct aacgcaaacc caaatgcaaa 1680 ccccaatgca aacccgaacg ccaatcccaa cgccaatcct aatgccaaca agaacaatca 1740 gggcaacggc cagggccaca acatgcccaa cgaccccaac cggaacgtgg acgagaacgc 1800 caacgccaac agcgccgtga agaacaacaa caacgaggag cccagcgaca agcacatcaa 1860 ggagtacctg aacaagatcc agaacagcct gagcaccgag tggagcccct gcagcgtgac 1920 ctgcggcaac ggcattcagg tgcggatcaa gcccggcagc gccaacaagc ccaaggacga 1980 gctggactac gccaatgaca tcgagaagaa gatctgcaag atggagaagt gcagcagcgt 2040 gttcaacgtg gtgaactcct gaggatccga tctttttccc tctgccaaaa attatgggga 2100 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc 2160 aatagtgtgt tggaattttt tgtgtctctc actcggaagc gatctgaatt catctatgtc 2220 gggtgcggag aaagaggtaa tgaaatggca 2250 <210> 18 <211> 2281 <212> DNA <213> Artificial Sequence <220> <223> Ade2 expression cassette <400> 18 taactataac ggtcctaagg tagcgaaagc tcagatcggc tgaccgccca acgacccccg 60 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 120 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 180 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 240 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc 300 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc 360 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 420 tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag 480 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg 540 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg 600 cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgcgg ccagctttat 660 tgcggtagtt tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc 720 gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag 780 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt 840 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca 900 ctcccagttc aattacagct cttaaggcta gagtggccgc accatgatga gaaaacttgc 960 catcctcagc gtcagctctt tcctgttcgt ggaggccctc ttccaggagt atcagtgcta 1020 cggaagcagc agcaatacaa gggtcctgaa cgagctcaac tatgacaacg ctggaacgaa 1080 cctgtataac gagctggaga tgaactacta tggcaagcag gagaactggt atagcctgaa 1140 gaagaacagc cggtccctgg gcgagaacga cgacggcaac aacaacaacg gcgacaacgg 1200 cagggagggc aaagatgagg acaagaggga cgggaacaac gaggataacg agaagctgcg 1260 gaagcccaag cacaagaaac tcaagcagcc cgccgacggg aacccggacc ccaatgcaaa 1320 tcccaacgtc gacccaaacg caaaccctaa cgtggacccc aacgccaatc ccaacgtcga 1380 tcctaatgcc aatccaaatg ccaaccctaa cgcaaatcct aatgcaaacc ccaacgccaa 1440 tcctaacgcc aacccaaatg ccaacccaaa cgctaacccc aacgctaacc caaatgcaaa 1500 tcccaatgct aacccaaacg tggaccctaa cgctaacccc aacgcaaacc ctaacgccaa 1560 tcctaacgca aaccccaatg caaacccaaa cgcaaatccc aacgctaacc ctaacgcaaa 1620 ccccaacgcc aaccctaatg ccaaccccaa tgctaacccc aacgccaatc caaacgcaaa 1680 tccaaacgcc aacccaaatg caaaccccaa cgctaatccc aacgccaacc caaacgccaa 1740 tcctaacaag aacaatcagg gcaacgggca gggccataac atgccgaacg accctaaccg 1800 gaatgtggac gagaacgcca acgccaacag cgccgtgaag aacaacaaca acgaggagcc 1860 ctccgacaag cacatcaagg aatacctgaa caagatccag aacagtctga gcaccgagtg 1920 gtccccctgc tccgtgacct gcggcaacgg catccaggtg aggatcaagc ccggctccgc 1980 caacaagccc aaggacgagc tggactacgc caacgacatc gagaagaaga tctgcaagat 2040 ggagaaatgc agctctgtgt tcaacgtcgt gaactccgcc atcggcctgt gaggatccga 2100 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc 2160 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc 2220 actcggaagc gatctgaatt catctatgtc gggtgcggag aaagaggtaa tgaaatggca 2280 t 2281 <210> 19 <211> 38398 <212> DNA <213> Artificial Sequence <220> <223> Synthetic recombinant vector C7-Ade2. <400> 19 catcatcaat aatatacctc aaacttttgg tgcgcgttaa tatgcaaatg agctgtttga 60 atttggggag ggaggaaggt gattggccga gagacgggcg accgttaggg gcggggcggg 120 tgacgttttg atgacgtggc cgtgaggcgg agccggtttg caagttctcg tgggaaaagt 180 gacgtcaaac gaggtgtggt ttgaacacgg aaatactcaa ttttcccgcg ctctctgaca 240 ggaaatgagg tgtttctggg cggatgcaag tgaaaacggg ccattttcgc gcgaaaactg 300 aatgaggaag tgaaaatctg agtaatttcg cgtttatggc agggaggagt atttgccgag 360 ggccgagtag actttgaccg attacgtggg ggtttcgatt accgtatttt tcacctaaat 420 ttccgcgtac ggtgtcaaag tccggtgttt ttacatcatt tccccgaaaa gtgccacctg 480 acgtaactat aacggtccta aggtagcgaa agctcagatc ggctgaccgc ccaacgaccc 540 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca 600 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta 660 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta 720 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat 780 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga 840 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca 900 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg 960 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc 1020 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct 1080 ccgcggccgg gaacggtgca ttggaacgcg gattccccgt gccaagagtg cggccagctt 1140 tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg acacaacagt 1200 ctcgaactta agctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac 1260 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg 1320 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt 1380 ccactcccag ttcaattaca gctcttaagg ctagagtggc cgcaccatga tgagaaaact 1440 tgccatcctc agcgtcagct ctttcctgtt cgtggaggcc ctcttccagg agtatcagtg 1500 ctacggaagc agcagcaata caagggtcct gaacgagctc aactatgaca acgctggaac 1560 gaacctgtat aacgagctgg agatgaacta ctatggcaag caggagaact ggtatagcct 1620 gaagaagaac agccggtccc tgggcgagaa cgacgacggc aacaacaaca acggcgacaa 1680 cggcagggag ggcaaagatg aggacaagag ggacgggaac aacgaggata acgagaagct 1740 gcggaagccc aagcacaaga aactcaagca gcccgccgac gggaacccgg accccaatgc 1800 aaatcccaac gtcgacccaa acgcaaaccc taacgtggac cccaacgcca atcccaacgt 1860 cgatcctaat gccaatccaa atgccaaccc taacgcaaat cctaatgcaa accccaacgc 1920 caatcctaac gccaacccaa atgccaaccc aaacgctaac cccaacgcta acccaaatgc 1980 aaatcccaat gctaacccaa acgtggaccc taacgctaac cccaacgcaa accctaacgc 2040 caatcctaac gcaaacccca atgcaaaccc aaacgcaaat cccaacgcta accctaacgc 2100 aaaccccaac gccaacccta atgccaaccc caatgctaac cccaacgcca atccaaacgc 2160 aaatccaaac gccaacccaa atgcaaaccc caacgctaat cccaacgcca acccaaacgc 2220 caatcctaac aagaacaatc agggcaacgg gcagggccat aacatgccga acgaccctaa 2280 tcggaatgtg gacgagaacg ccaacgccaa cagcgccgtg aagaacaaca acaacgagga 2340 gccctccgac aagcacatca aggaatacct gaacaagatc cagaacagtc tgagcaccga 2400 gtggtccccc tgctccgtga cctgcggcaa cggcatccag gtgaggatca agcccggctc 2460 cgccaacaag cccaaggacg agctggacta cgccaacgac atcgagaaga agatctgcaa 2520 gatggagaaa tgcagctctg tgttcaacgt cgtgaactcc gccatcggcc tgtgaggatc 2580 cgatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac 2640 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct 2700 ctcactcgga agcgatctga attcatctat gtcgggtgcg gagaaagagg taatgaaatg 2760 gcattatggg tattatgggt ctgcattaat gaatcggcca gatatgctgg ccaccgtgca 2820 tgtgacctcg cacccccgca agacatggcc cgagttcgag cacaacgtca tgacccgatg 2880 caatgtgcac ctggggtccc gccgaggcat gttcatgccc taccagtgca acatgcaatt 2940 tgtgaaggtg ctgctggagc ccgatgccat gtccagagtg agcctgacgg gggtgtttga 3000 catgaatgtg gagctgtgga aaattctgag atatgatgaa tccaagacca ggtgccgggc 3060 ctgcgaatgc ggaggcaagc acgccaggct tcagcccgtg tgtgtggagg tgacggagga 3120 cctgcgaccc gatcatttgg tgttgtcctg caacgggacg gagttcggct ccagcgggga 3180 agaatctgac tagagtgagt agtgtttggg ggaggtggag ggcttgtatg aggggcagaa 3240 tgactaaaat ctgtgttttt ctgtgtgttg cagcagcatg agcggaagcg cctcctttga 3300 gggaggggta ttcagccctt atctgacggg gcgtctcccc tcctgggcgg gagtgcgtca 3360 gaatgtgatg ggatccacgg tggacggccg gcccgtgcag cccgcgaact cttcaaccct 3420 gacctacgcg accctgagct cctcgtccgt ggacgcagct gccgccgcag ctgctgcttc 3480 cgccgccagc gccgtgcgcg gaatggccct gggcgccggc tactacagct ctctggtggc 3540 caactcgact tccaccaata atcccgccag cctgaacgag gagaagctgc tgctgctgat 3600 ggcccagctc gaggccctga cccagcgcct gggcgagctg acccagcagg tggctcagct 3660 gcaggcggag acgcgggccg cggttgccac ggtgaaaacc aaataaaaaa tgaatcaata 3720 aataaacgga gacggttgtt gattttaaca cagagtcttg aatctttatt tgatttttcg 3780 cgcgcggtag gccctggacc accggtctcg atcattgagc acccggtgga ttttttccag 3840 gacccggtag aggtgggctt ggatgttgag gtacatgggc atgagcccgt cccgggggtg 3900 gaggtagctc cattgcaggg cctcgtgctc gggggtggtg ttgtaaatca cccagtcata 3960 gcaggggcgc agggcgtggt gctgcacgat gtccttgagg aggagactga tggccacggg 4020 cagccccttg gtgtaggtgt tgacgaacct gttgagctgg gagggatgca tgcgggggga 4080 gatgagatgc atcttggcct ggatcttgag attggcgatg ttcccgccca gatcccgccg 4140 ggggttcatg ttgtgcagga ccaccagcac ggtgtatccg gtgcacttgg ggaatttgtc 4200 atgcaacttg gaagggaagg cgtgaaagaa tttggagacg cccttgtgac cgcccaggtt 4260 ttccatgcac tcatccatga tgatggcgat gggcccgtgg gcggcggcct gggcaaagac 4320 gtttcggggg tcggacacat cgtagttgtg gtcctgggtg agctcgtcat aggccatttt 4380 aatgaatttg gggcggaggg tgcccgactg ggggacgaag gtgccctcga tcccgggggc 4440 gtagttgccc tcgcagatct gcatctccca ggccttgagc tcggaggggg ggatcatgtc 4500 cacctgcggg gcgatgaaaa aaacggtttc cggggcgggg gagatgagct gggccgaaag 4560 caggttccgg agcagctggg acttgccgca gccggtgggg ccgtagatga ccccgatgac 4620 cggctgcagg tggtagttga gggagagaca gctgccgtcc tcgcggagga ggggggccac 4680 ctcgttcatc atctcgcgca catgcatgtt ctcgcgcacg agttccgcca ggaggcgctc 4740 gccccccagc gagaggagct cttgcagcga ggcgaagttt ttcagcggct tgagcccgtc 4800 ggccatgggc attttggaga gggtctgttg caagagttcc agacggtccc agagctcggt 4860 gatgtgctct agggcatctc gatccagcag acctcctcgt ttcgcgggtt ggggcgactg 4920 cgggagtagg gcaccaggcg atgggcgtcc agcgaggcca gggtccggtc cttccagggt 4980 cgcagggtcc gcgtcagcgt ggtctccgtc acggtgaagg ggtgcgcgcc gggctgggcg 5040 cttgcgaggg tgcgcttcag gctcatccgg ctggtcgaga accgctcccg gtcggcgccc 5100 tgcgcgtcgg ccaggtagca attgagcatg agttcgtagt tgagcgcctc ggccgcgtgg 5160 cccttggcgc ggagcttacc tttggaagtg tgtccgcaga cgggacagag gagggacttg 5220 agggcgtaga gcttgggggc gaggaagacg gactcggggg cgtaggcgtc cgcgccgcag 5280 ctggcgcaga cggtctcgca ctccacgagc caggtgaggt cgggccggtt ggggtcaaaa 5340 acgaggtttc ctccgtgctt tttgatgcgt ttcttacctc tggtctccat gagctcgtgt 5400 ccccgctggg tgacaaagag gctgtccgtg tccccgtaga ccgactttat gggccggtcc 5460 tcgagcgggg tgccgcggtc ctcgtcgtag aggaaccccg cccactccga gacgaaggcc 5520 cgggtccagg ccagcacgaa ggaggccacg tgggaggggt agcggtcgtt gtccaccagc 5580 gggtccacct tctccagggt atgcaagcac atgtccccct cgtccacatc caggaaggtg 5640 attggcttgt aagtgtaggc cacgtgaccg ggggtcccgg ccgggggggt ataaaagggg 5700 gcgggcccct gctcgtcctc actgtcttcc ggatcgctgt ccaggagcgc cagctgttgg 5760 ggtaggtatt ccctctcgaa ggctggcata acctcggcac tcaggttgtc agtttctaga 5820 aacgaggagg atttgatatt gacggtgccg ttggagacgc ctttcatgag cccctcgtcc 5880 atctggtcag aaaagacgat ctttttgttg tcgagcttgg tggcgaagga gccgtagagg 5940 gcgttggaga ggagcttggc gatggagcgc atggtctggt tcttttcctt gtcggcgcgc 6000 tccttggcgg cgatgttgag ctgcacgtac tcgcgcgcca cgcacttcca ttcggggaag 6060 acggtggtga gctcgtcggg cacgattctg acccgccagc cgcggttgtg cagggtgatg 6120 aggtccacgc tggtggccac ctcgccgcgc aggggctcgt tggtccagca gaggcgcccg 6180 cccttgcgcg agcagaaggg gggcagcggg tccagcatga gctcgtcggg ggggtcggcg 6240 tccacggtga agatgccggg cagaagctcg gggtcgaagt agctgatgca ggtgtccaga 6300 tcgtccagcg ccgcttgcca gtcgcgcacg gccagcgcgc gctcgtaggg gctgaggggc 6360 gtgccccagg gcatggggtg cgtgagcgcg gaggcgtaca tgccgcagat gtcgtagacg 6420 tagaggggct cctcgaggac gccgatgtag gtggggtagc agcgcccccc gcggatgctg 6480 gcgcgcacgt agtcgtacag ctcgtgcgag ggcgcgagga gccccgtgcc gaggttggag 6540 cgttgcggct tttcggcgcg gtagacgatc tggcggaaga tggcgtggga gttggaggag 6600 atggtgggcc tctggaagat gttgaagtgg gcgtggggca ggccgaccga gtccctgatg 6660 aagtgggcgt aggagtcctg cagcttggcg acgagctcgg cggtgacgag gacgtccagg 6720 gcgcagtagt cgagggtctc ttggatgatg tcgtacttga gctggccctt ctgcttccac 6780 agctcgcggt tgagaaggaa ctcttcgcgg tccttccagt actcttcgag ggggaacccg 6840 tcctgatcgg cacggtaaga gcccaccatg tagaactggt tgacggcctt gtaggcgcag 6900 cagcccttct ccacggggag ggcgtaagct tgtgcggcct tgcgcaggga ggtgtgggtg 6960 agggcgaagg tgtcgcgcac catgaccttg aggaactggt gcttgaagtc gaggtcgtcg 7020 cagccgccct gctcccagag ctggaagtcc gtgcgcttct tgtaggcggg gttgggcaaa 7080 gcgaaagtaa catcgttgaa gaggatcttg cccgcgcggg gcatgaagtt gcgagtgatg 7140 cggaaaggct ggggcacctc ggcccggttg ttgatgacct gggcggcgag gacgatctcg 7200 tcgaagccgt tgatgttgtg cccgacgatg tagagttcca cgaatcgcgg gcggccctta 7260 acgtggggca gcttcttgag ctcgtcgtag gtgagctcgg cggggtcgct gagcccgtgc 7320 tgctcgaggg cccagtcggc gacgtggggg ttggcgctga ggaaggaagt ccagagatcc 7380 acggccaggg cggtctgcaa gcggtcccgg tactgacgga actgctggcc cacggccatt 7440 ttttcggggg tgacgcagta gaaggtgcgg gggtcgccgt gccagcggtc ccacttgagc 7500 tggagggcga ggtcgtgggc gagctcgacg agcggcgggt ccccggagag tttcatgacc 7560 agcatgaagg ggacgagctg cttgccgaag gaccccatcc aggtgtaggt ttccacatcg 7620 taggtgagga agagcctttc ggtgcgagga tgcgagccga tggggaagaa ctggatctcc 7680 tgccaccagt tggaggaatg gctgttgatg tgatggaagt agaaatgccg acggcgcgcc 7740 gagcactcgt gcttgtgttt atacaagcgt ccgcagtgct cgcaacgctg cacgggatgc 7800 acgtgctgca cgagctgtac ctgggttcct ttgacgagga atttcagtgg gcagtggagc 7860 gctggcggct gcatctggtg ctgtactacg tcctggccat cggcgtggcc atcgtctgcc 7920 tcgatggtgg tcatgctgac gagcccgcgc gggaggcagg tccagacttc ggctcggacg 7980 ggtcggagag cgaggacgag ggcgcgcagg ccggagctgt ccagggtcct gagacgctgc 8040 ggagtcaggt cagtgggcag cggcggcgcg cggttgactt gcaggagctt ttccagggcg 8100 cgcgggaggt ccagatggta cttgatctcc acggcgccgt tggtggcgac gtccacggct 8160 tgcagggtcc cgtgcccctg gggcgccacc accgtgcccc gtttcttctt gggcgctgct 8220 tccatgccgg tcagaagcgg cggcgaggac gcgcgccggg cggcaggggc ggctcgggac 8280 ccggaggcag gggcggcagg ggcacgtcgg cgccgcgcgc gggcaggttc tggtactgcg 8340 cccggagaag actggcgtga gcgacgacgc gacggttgac gtcctggatc tgacgcctct 8400 gggtgaaggc cacgggaccc gtgagtttga acctgaaaga gagttcgaca gaatcaatct 8460 cggtatcgtt gacggcggcc tgccgcagga tctcttgcac gtcgcccgag ttgtcctggt 8520 aggcgatctc ggtcatgaac tgctcgatct cctcctcctg aaggtctccg cggccggcgc 8580 gctcgacggt ggccgcgagg tcgttggaga tgcggcccat gagctgcgag aaggcgttca 8640 tgccggcctc gttccagacg cggctgtaga ccacggctcc gtcggggtcg cgcgcgcgca 8700 tgaccacctg ggcgaggttg agctcgacgt ggcgcgtgaa gaccgcgtag ttgcagaggc 8760 gctggtagag gtagttgagc gtggtggcga tgtgctcggt gacgaagaag tacatgatcc 8820 agcggcggag cggcatctcg ctgacgtcgc ccagggcttc caagcgctcc atggcctcgt 8880 agaagtccac ggcgaagttg aaaaactggg agttgcgcgc cgagacggtc aactcctcct 8940 ccagaagacg gatgagctca gcgatggtgg cgcgcacctc gcgctcgaag gccccggggg 9000 gctcctcttc ttccatctct tcctcctcca ctaacatctc ttctacttcc tcctcaggag 9060 gcggcggcgg gggaggggcc ctgcgtcgcc ggcggcgcac gggcagacgg tcgatgaagc 9120 gctcgatggt ctccccgcgc cggcgacgca tggtctcggt gacggcgcgc ccgtcctcgc 9180 ggggccgcag cgtgaagacg ccgccgcgca tctccaggtg gccgccgggg gggtctccgt 9240 tgggcaggga gagggcgctg acgatgcatc ttatcaattg gcccgtaggg actccgcgca 9300 aggacctgag cgtctcgaga tccacgggat ccgaaaaccg ctgaacgaag gcttcgagcc 9360 agtcgcagtc gcaaggtagg ctgagcccgg tttcttgttc ttcggggatt tcgggaggcg 9420 ggcgggcgat gctgctggtg atgaagttga agtaggcggt cctgagacgg cggatggtgg 9480 cgaggagcac caggtccttg ggcccggctt gctggatgcg cagacggtcg gccatgcccc 9540 aggcgtggtc ctgacacctg gcgaggtcct tgtagtagtc ctgcatgagc cgctccacgg 9600 gcacctcctc ctcgcccgcg cggccgtgca tgcgcgtgag cccgaacccg cgctggggct 9660 ggacgagcgc caggtcggcg acgacgcgct cggcgaggat ggcctgctgt atctgggtga 9720 gggtggtctg gaagtcgtcg aagtcgacga agcggtggta ggctccggtg ttgatggtat 9780 aggagcagtt ggccatgacg gaccagttga cggtctggtg gccgggtcgc acgagctcgt 9840 ggtacttgag gcgcgagtag gcgcgcgtgt cgaagatgta gtcgttgcag gtgcgcacga 9900 ggtactggta tccgacgagg aagtgcggcg gcggctggcg gtagagcggc catcgctcgg 9960 tggcgggggc gccgggcgcg aggtcctcga gcatgaggcg gtggtagccg tagatgtacc 10020 tggacatcca ggtgatgccg gcggcggtgg tggaggcgcg cgggaactcg cggacgcggt 10080 tccagatgtt gcgcagcggc aggaagtagt tcatggtggc cgcggtctgg cccgtgaggc 10140 gcgcgcagtc gtggatgctc tagacatacg ggcaaaaacg aaagcggtca gcggctcgac 10200 tccgtggcct ggaggctaag cgaacgggtt gggctgcgcg tgtaccccgg ttcgaatctc 10260 gaatcaggct ggagccgcag ctaacgtggt actggcactc ccgtctcgac ccaagcctgc 10320 taacgaaacc tccaggatac ggaggcgggt cgttttttgg ccttggtcgc tggtcatgaa 10380 aaactagtaa gcgcggaaag cgaccgcccg cgatggctcg ctgccgtagt ctggagaaag 10440 aatcgccagg gttgcgttgc ggtgtgcccc ggttcgagcc tcagcgctcg gcgccggccg 10500 gattccgcgg ctaacgtggg cgtggctgcc ccgtcgtttc caagacccct tagccagccg 10560 acttctccag ttacggagcg agcccctctt tttcttgtgt ttttgccaga tgcatcccgt 10620 actgcggcag atgcgccccc accctccacc tcaaccgccc ctaccgccgc agcagcagca 10680 acagccggcg cttctgcccc cgccccagca gcagccagcc actaccgcgg cggccgccgt 10740 gagcggagcc ggcgttcagt atgacctggc cttggaagag ggcgaggggc tggcgcggct 10800 gggggcgtcg tcgccggagc ggcacccgcg cgtgcagatg aaaagggacg ctcgcgaggc 10860 ctacgtgccc aagcagaacc tgttcagaga caggagcggc gaggagcccg aggagatgcg 10920 cgcctcccgc ttccacgcgg ggcgggagct gcggcgcggc ctggaccgaa agcgggtgct 10980 gagggacgag gatttcgagg cggacgagct gacggggatc agccccgcgc gcgcgcacgt 11040 ggccgcggcc aacctggtca cggcgtacga gcagaccgtg aaggaggaga gcaacttcca 11100 aaaatccttc aacaaccacg tgcgcacgct gatcgcgcgc gaggaggtga ccctgggcct 11160 gatgcacctg tgggacctgc tggaggccat cgtgcagaac cccacgagca agccgctgac 11220 ggcgcagctg tttctggtgg tgcagcacag tcgggacaac gagacgttca gggaggcgct 11280 gctgaatatc accgagcccg agggccgctg gctcctggac ctggtgaaca ttctgcagag 11340 catcgtggtg caggagcgcg ggctgccgct gtccgagaag ctggcggcta tcaacttctc 11400 ggtgctgagc ctgggcaagt actacgctag gaagatctac aagaccccgt acgtgcccat 11460 agacaaggag gtgaagatcg acgggtttta catgcgcatg accctgaaag tgctgaccct 11520 gagcgacgat ctgggggtgt accgcaacga caggatgcac cgcgcggtga gcgccagccg 11580 ccggcgcgag ctgagcgacc aggagctgat gcacagcctg cagcgggccc tgaccggggc 11640 cgggaccgag ggggagagct actttgacat gggcgcggac ctgcgctggc agcccagccg 11700 ccgggccttg gaagctgccg gcggttcccc ctacgtggag gaggtggacg atgaggagga 11760 ggagggcgag tacctggaag actgatggcg cgaccgtatt tttgctagat gcagcaacag 11820 ccaccgcctc ctgatcccgc gatgcgggcg gcgctgcaga gccagccgtc cggcattaac 11880 tcctcggacg attggaccca ggccatgcaa cgcatcatgg cgctgacgac ccgcaatccc 11940 gaagccttta gacagcagcc tcaggccaac cggctctcgg ccatcctgga ggccgtggtg 12000 ccctcgcgct cgaaccccac gcacgagaag gtgctggcca tcgtgaacgc gctggtggag 12060 aacaaggcca tccgcggcga cgaggccggg ctggtgtaca acgcgctgct ggagcgcgtg 12120 gcccgctaca acagcaccaa cgtgcagacg aacctggacc gcatggtgac cgacgtgcgc 12180 gaggcggtgt cgcagcgcga gcggttccac cgcgagtcga acctgggctc catggtggcg 12240 ctgaacgcct tcctgagcac gcagcccgcc aacgtgcccc ggggccagga ggactacacc 12300 aacttcatca gcgcgctgcg gctgatggtg gccgaggtgc cccagagcga ggtgtaccag 12360 tcggggccgg actacttctt ccagaccagt cgccagggct tgcagaccgt gaacctgagc 12420 caggctttca agaacttgca gggactgtgg ggcgtgcagg ccccggtcgg ggaccgcgcg 12480 acggtgtcga gcctgctgac gccgaactcg cgcctgctgc tgctgctggt ggcgcccttc 12540 acggacagcg gcagcgtgag ccgcgactcg tacctgggct acctgcttaa cctgtaccgc 12600 gaggccatcg ggcaggcgca cgtggacgag cagacctacc aggagatcac ccacgtgagc 12660 cgcgcgctgg gccaggagga cccgggcaac ctggaggcca ccctgaactt cctgctgacc 12720 aaccggtcgc agaagatccc gccccagtac gcgctgagca ccgaggagga gcgcatcctg 12780 cgctacgtgc agcagagcgt ggggctgttc ctgatgcagg agggggccac gcccagcgcc 12840 gcgctcgaca tgaccgcgcg caacatggag cccagcatgt acgctcgcaa ccgcccgttc 12900 atcaataagc tgatggacta cttgcatcgg gcggccgcca tgaactcgga ctactttacc 12960 aacgccatct tgaacccgca ctggctcccg ccgcccgggt tctacacggg cgagtacgac 13020 atgcccgacc ccaacgacgg gttcctgtgg gacgacgtgg acagcagcgt gttctcgccg 13080 cgccccgcca ccaccgtgtg gaagaaagag ggcggggacc ggcggccgtc ctcggcgctg 13140 tccggtcgcg cgggtgctgc cgcggcggtg cctgaggccg ccagcccctt cccgagcctg 13200 cccttttcgc tgaacagcgt gcgcagcagc gagctgggtc ggctgacgcg gccgcgcctg 13260 ctgggcgagg aggagtacct gaacgactcc ttgttgaggc ccgagcgcga gaagaacttc 13320 cccaataacg ggatagagag cctggtggac aagatgagcc gctggaagac gtacgcgcac 13380 gagcacaggg acgagccccg agctagcagc agcgcaggca cccgtagacg ccagcgacac 13440 gacaggcagc ggggtctggt gtgggacgat gaggattccg ccgacgacag cagcgtgttg 13500 gacttgggtg ggagtggtgg tggtaacccg ttcgctcact tgcgcccccg tatcgggcgc 13560 ctgatgtaag aatctgaaaa aataaaaaac ggtactcacc aaggccatgg cgaccagcgt 13620 gcgttcttct ctgttgtttg tagtagtatg atgaggcgcg tgtacccgga gggtcctcct 13680 ccctcgtacg agagcgtgat gcagcaggcg gtggcggcgg cgatgcagcc cccgctggag 13740 gcgccttacg tgcccccgcg gtacctggcg cctacggagg ggcggaacag cattcgttac 13800 tcggagctgg cacccttgta cgataccacc cggttgtacc tggtggacaa caagtcggcg 13860 gacatcgcct cgctgaacta ccagaacgac cacagcaact tcctgaccac cgtggtgcag 13920 aacaacgatt tcacccccac ggaggccagc acccagacca tcaactttga cgagcgctcg 13980 cggtggggcg gccagctgaa aaccatcatg cacaccaaca tgcccaacgt gaacgagttc 14040 atgtacagca acaagttcaa ggcgcgggtg atggtctcgc gcaagacccc caatggggtc 14100 gcggtggatg agaattatga tggtagtcag gacgagctga cttacgagtg ggtggagttt 14160 gagctgcccg agggcaactt ctcggtgacc atgaccatcg atctgatgaa caacgccatc 14220 atcgacaact acttggcggt ggggcgtcag aacggggtgc tggagagcga catcggcgtg 14280 aagttcgaca cgcgcaactt ccggctgggc tgggaccccg tgaccgagct ggtgatgccg 14340 ggcgtgtaca ccaacgaggc cttccacccc gacatcgtcc tgctgcccgg ctgcggcgtg 14400 gacttcaccg agagccgcct cagcaacctg ctgggcatcc gcaagcggca gcccttccag 14460 gagggcttcc agatcctgta cgaggacctg gaggggggca acatccccgc gctcttggat 14520 gtcgaagcct atgagaaaag caaggaggag gccgccgcag cggcgaccgc agccgtggcc 14580 accgcctcta ccgaggtgcg gggcgataat tttgctagcg ccgcggcagt ggccgaggcg 14640 gctgaaaccg aaagtaagat agtcatccag ccggtggaga aggacagcaa ggacaggagc 14700 tacaacgtgc tcgcggacaa gaaaaacacc gcctaccgca gctggtacct ggcctacaac 14760 tacggcgacc ccgagaaggg cgtgcgctcc tggacgctgc tcaccacctc ggacgtcacc 14820 tgcggcgtgg agcaagtcta ctggtcgctg cccgacatga tgcaagaccc ggtcaccttc 14880 cgctccacgc gtcaagttag caactacccg gtggtgggcg ccgagctcct gcccgtctac 14940 tccaagagct tcttcaacga gcaggccgtc tactcgcagc agctgcgcgc cttcacctcg 15000 ctcacgcacg tcttcaaccg cttccccgag aaccagatcc tcgtccgccc gcccgcgccc 15060 accattacca ccgtcagtga aaacgttcct gctctcacag atcacgggac cctgccgctg 15120 cgcagcagta tccggggagt ccagcgcgtg accgtcactg acgccagacg ccgcacctgc 15180 ccctacgtct acaaggccct gggcgtagtc gcgccgcgcg tcctctcgag ccgcaccttc 15240 taaaaaatgt ccattctcat ctcgcccagt aataacaccg gttggggcct gcgcgcgccc 15300 agcaagatgt acggaggcgc tcgccaacgc tccacgcaac accccgtgcg cgtgcgcggg 15360 cacttccgcg ctccctgggg cgccctcaag ggccgcgtgc gctcgcgcac caccgtcgac 15420 gacgtgatcg accaggtggt ggccgacgcg cgcaactaca cgcccgccgc cgcgcccgcc 15480 tccaccgtgg acgccgtcat cgacagcgtg gtggccgatg cgcgccggta cgcccgcgcc 15540 aagagccggc ggcggcgcat cgcccggcgg caccggagca cccccgccat gcgcgcggcg 15600 cgagccttgc tgcgcagggc caggcgcacg ggacgcaggg ccatgctcag ggcggccaga 15660 cgcgcggcct ccggcagcag cagcgccggc aggacccgca gacgcgcggc cacggcggcg 15720 gcggcggcca tcgccagcat gtcccgcccg cggcgcggca acgtgtactg ggtgcgcgac 15780 gccgccaccg gtgtgcgcgt gcccgtgcgc acccgccccc ctcgcacttg aagatgctga 15840 cttcgcgatg ttgatgtgtc ccagcggcga ggaggatgtc caagcgcaaa tacaaggaag 15900 agatgctcca ggtcatcgcg cctgagatct acggccccgc ggtgaaggag gaaagaaagc 15960 cccgcaaact gaagcgggtc aaaaaggaca aaaaggagga ggaagatgtg gacggactgg 16020 tggagtttgt gcgcgagttc gccccccggc ggcgcgtgca gtggcgcggg cggaaagtga 16080 aaccggtgct gcggcccggc accacggtgg tcttcacgcc cggcgagcgt tccggctccg 16140 cctccaagcg ctcctacgac gaggtgtacg gggacgagga catcctcgag caggcggtcg 16200 agcgtctggg cgagtttgct tacggcaagc gcagccgccc cgcgcccttg aaagaggagg 16260 cggtgtccat cccgctggac cacggcaacc ccacgccgag cctgaagccg gtgaccctgc 16320 agcaggtgct gccgagcgcg gcgccgcgcc ggggcttcaa gcgcgagggc ggcgaggatc 16380 tgtacccgac catgcagctg atggtgccca agcgccagaa gctggaggac gtgctggagc 16440 acatgaaggt ggaccccgag gtgcagcccg aggtcaaggt gcggcccatc aagcaggtgg 16500 ccccgggcct gggcgtgcag accgtggaca tcaagatccc cacggagccc atggaaacgc 16560 agaccgagcc cgtgaagccc agcaccagca ccatggaggt gcagacggat ccctggatgc 16620 cggcgccggc ttccaccact cgccgaagac gcaagtacgg cgcggccagc ctgctgatgc 16680 ccaactacgc gctgcatcct tccatcatcc ccacgccggg ctaccgcggc acgcgcttct 16740 accgcggcta caccagcagc cgccgcaaga ccaccacccg ccgccgccgt cgtcgcaccc 16800 gccgcagcag caccgcgact tccgccgccg ccctggtgcg gagagtgtac cgcagcgggc 16860 gcgagcctct gaccctgccg cgcgcgcgct accacccgag catcgccatt taactctgcc 16920 gtcgcctcct acttgcagat atggccctca catgccgcct ccgcgtcccc attacgggct 16980 accgaggaag aaagccgcgc cgtagaaggc tgacggggaa cgggctgcgt cgccatcacc 17040 accggcggcg gcgcgccatc agcaagcggt tggggggagg cttcctgccc gcgctgatcc 17100 ccatcatcgc cgcggcgatc ggggcgatcc ccggcatagc ttccgtggcg gtgcaggcct 17160 ctcagcgcca ctgagacaca gcttggaaaa tttgtaataa aaaaatggac tgacgctcct 17220 ggtcctgtga tgtgtgtttt tagatggaag acatcaattt ttcgtccctg gcaccgcgac 17280 acggcacgcg gccgtttatg ggcacctgga gcgacatcgg caacagccaa ctgaacgggg 17340 gcgccttcaa ttggagcagt ctctggagcg ggcttaagaa tttcgggtcc acgctcaaaa 17400 cctatggcaa caaggcgtgg aacagcagca cagggcaggc gctgagggaa aagctgaaag 17460 agcagaactt ccagcagaag gtggtcgatg gcctggcctc gggcatcaac ggggtggtgg 17520 acctggccaa ccaggccgtg cagaaacaga tcaacagccg cctggacgcg gtcccgcccg 17580 cggggtccgt ggagatgccc caggtggagg aggagctgcc tcccctggac aagcgcggcg 17640 acaagcgacc gcgtcccgac gcggaggaga cgctgctgac gcacacggac gagccgcccc 17700 cgtacgagga ggcggtgaaa ctgggtctgc ccaccacgcg gcccgtggcg cctctggcca 17760 ccggggtgct gaaacccagc agcagcagcc agcccgcgac cctggacttg cctccgcctg 17820 cttcccgccc ctccacagtg gctaagcccc tgccgccggt ggccgtcgcg tcgcgcgccc 17880 cccgaggccg cccccaggcg aactggcaga gcactctgaa cagcatcgtg ggtctgggag 17940 tgcagagtgt gaagcgccgc cgctgctatt aaaagacact gtagcgctta acttgcttgt 18000 ctgtgtgtat atgtatgtcc gccgaccaga aggaggaaga ggcgcgtcgc cgagttgcaa 18060 gatggccacc ccatcgatgc tgccccagtg ggcgtacatg cacatcgccg gacaggacgc 18120 ttcggagtac ctgagtccgg gtctggtgca gttcgcccgc gccacagaca cctacttcag 18180 tctggggaac aagtttagga accccacggt ggcgcccacg cacgatgtga ccaccgaccg 18240 cagccagcgg ctgacgctgc gcttcgtgcc cgtggaccgc gaggacaaca cctactcgta 18300 caaagtgcgc tacacgctgg ccgtgggcga caaccgcgtg ctggacatgg ccagcaccta 18360 ctttgacatc cgcggcgtgc tggatcgggg gcccagcttc aaaccctact ccggcaccgc 18420 ctacaacagc ctggctccca agggagcgcc caacacttgc cagtggacat ataaagctgg 18 480 tgatactgat acagaaaaaa cctatacata tggaaatgca cctgtgcaag gcattagcat 18540 tacaaaggat ggtattcaac ttggaactga cagcgatggt caggcaatct atgcagacga 18600 aacttatcaa ccagagcctc aagtgggtga tgctgaatgg catgacatca ctggtactga 18660 tgaaaaatat ggaggcagag ctcttaagcc tgacaccaaa atgaagcctt gctatggttc 18720 ttttgccaag cctaccaata aagaaggagg ccaggcaaat gtgaaaaccg aaacaggcgg 18780 taccaaagaa tatgacattg acatggcatt cttcgataat cgaagtgcag ctgccgccgg 18840 cctagcccca gaaattgttt tgtatactga gaatgtggat ctggaaactc cagataccca 18900 tattgtatac aaggcaggta cagatgacag tagctcttct atcaatttgg gtcagcagtc 18960 catgcccaac agacccaact acattggctt cagagacaac tttatcggtc tgatgtacta 19020 caacagcact ggcaatatgg gtgtactggc tggacaggcc tcccagctga atgctgtggt 19080 ggacttgcag gacagaaaca ccgaactgtc ctaccagctc ttgcttgact ctctgggtga 19140 cagaaccagg tatttcagta tgtggaatca ggcggtggac agttatgacc ccgatgtgcg 19200 cattattgaa aatcacggtg tggaggatga acttcctaac tattgcttcc ccctggatgc 19260 tgtgggtaga actgatactt accagggaat taaggccaat ggtgataatc aaaccacctg 19320 gaccaaagat gatactgtta atgatgctaa tgaattgggc aagggcaatc ctttcgccat 19380 ggagatcaac atccaggcca acctgtggcg gaacttcctc tacgcgaacg tggcgctgta 19440 cctgcccgac tcctacaagt acacgccggc caacatcacg ctgcccacca acaccaacac 19500 ctacgattac atgaacggcc gcgtggtggc gccctcgctg gtggacgcct acatcaacat 19560 cggggcgcgc tggtcgctgg accccatgga caacgtcaac cccttcaacc accaccgcaa 19620 cgcgggcctg cgataccgct ccatgctcct gggcaacggg cgctacgtgc ccttccacat 19680 ccaggtgccc caaaagtttt tcgccatcaa gagcctcctg ctcctgcccg ggtcctacac 19740 ctacgagtgg aacttccgca aggacgtcaa catgatcctg cagagctccc tcggcaacga 19800 cctgcgcacg gacggggcct ccatcgcctt caccagcatc aacctctacg ccaccttctt 19860 ccccatggcg cacaacaccg cctccacgct cgaggccatg ctgcgcaacg acaccaacga 19920 ccagtccttc aacgactacc tctcggcggc caacatgctc taccccatcc cggccaacgc 19980 caccaacgtg cccatctcca tcccctcgcg caactgggcc gccttccgcg gctggtcctt 20040 cacgcgcctc aagacccgcg agacgccctc gctcggctcc gggttcgacc cctacttcgt 20 100 ctactcgggc tccatcccct acctcgacgg caccttctac ctcaaccaca ccttcaagaa 20160 ggtctccatc accttcgact cctccgtcag ctggcccggc aacgaccgcc tcctgacgcc 20220 caacgagttc gaaatcaagc gcaccgtcga cggagagggg tacaacgtgg cccagtgcaa 20280 catgaccaag gactggttcc tggtccagat gctggcccac tacaacatcg gctaccaggg 20340 cttctacgtg cccgagggct acaaggaccg catgtactcc ttcttccgca acttccagcc 20400 catgagccgc caggtcgtgg acgaggtcaa ctacaaggac taccaggccg tcaccctggc 20460 ctaccagcac aacaactcgg gcttcgtcgg ctacctcgcg cccaccatgc gccagggcca 20520 gccctacccc gccaactacc cctacccgct catcggcaag agcgccgtcg ccagcgtcac 20580 ccagaaaaag ttcctctgcg accgggtcat gtggcgcatc cccttctcca gcaacttcat 20640 gtccatgggc gcgctcaccg acctcggcca gaacatgctc tacgccaact ccgcccacgc 20700 gctagacatg aatttcgaag tcgaccccat ggatgagtcc acccttctct atgttgtctt 20760 cgaagtcttc gacgtcgtcc gagtgcacca gccccaccgc ggcgtcatcg aggccgtcta 20820 cctgcgcacg cccttctcgg ccggcaacgc caccacctaa gcctcttgct tcttgcaaga 20880 tgacggcctg cgcgggctcc ggcgagcagg agctcagggc catcctccgc gacctgggct 20940 gcgggccctg cttcctgggc accttcgaca agcgcttccc gggattcatg gccccgcaca 21000 agctggcctg cgccatcgtc aacacggccg gccgcgagac cgggggcgag cactggctgg 21060 ccttcgcctg gaacccgcgc tcccacacct gctacctctt cgaccccttc gggttctcgg 21120 acgagcgcct caagcagatc taccagttcg agtacgaggg cctgctgcgt cgcagcgccc 21180 tggccaccga ggaccgctgc gtcaccctgg aaaagtccac ccagaccgtg cagggtccgc 21240 gctcggccgc ctgcgggctc ttctgctgca tgttcctgca cgccttcgtg cactggcccg 21300 accgccccat ggacaagaac cccaccatga acttgctgac gggggtgccc aacggcatgc 21360 tccagtcgcc ccaggtggaa cccaccctgc gccgcaacca ggaggcgctc taccgcttcc 21420 tcaacgccca ctccgcctac tttcgctccc accgcgcgcg catcgagaag gccaccgcct 21480 tcgaccgcat gaatcaagac atgtaatccg gtgtgtgtat gtgaatgctt tattcatcat 21540 aataaacagc acatgtttat gccaccttct ctgaggctct gactttattt agaaatcgaa 21600 ggggttctgc cggctctcgg catggcccgc gggcagggat acgttgcgga actggtactt 21660 gggcagccac ttgaactcgg ggatcagcag cttcggcacg gggaggtcgg ggaacgagtc 21720 gctccacagc ttgcgcgtga gttgcagggc gcccagcagg tcgggcgcgg agatcttgaa 21780 atcgcagttg ggacccgcgt tctgcgcgcg agagttacgg tacacggggt tgcagcactg 21840 gaacaccatc agggccgggt gcttcacgct cgccagcacc gtcgcgtcgg tgatgccctc 21900 cacgtccaga tcctcggcgt tggccatccc gaagggggtc atcttgcagg tctgccgccc 21960 catgctgggc acgcagccgg gcttgtggtt gcaatcgcag tgcaggggga tcagcatcat 22020 ctgggcctgc tcggagctca tgcccgggta catggccttc atgaaagcct ccagctggcg 22080 gaaggcctgc tgcgccttgc cgccctcggt gaagaagacc ccgcaggact tgctagagaa 22140 ctggttggtg gcgcagccag cgtcgtgcac gcagcagcgc gcgtcgttgt tggccagctg 22200 caccacgctg cgcccccagc ggttctgggt gatcttggcc cggtcggggt tctccttcag 22260 cgcgcgctgc ccgttctcgc tcgccacatc catctcgatc gtgtgctcct tctggatcat 22320 cacggtcccg tgcaggcacc gcagcttgcc ctcggcctcg gtgcacccgt gcagccacag 22380 cgcgcagccg gtgctctccc agttcttgtg ggcgatctgg gagtgcgagt gcacgaagcc 22440 ctgcaggaag cggcccatca tcgtggtcag ggtcttgttg ctggtgaagg tcagcggaat 22500 gccgcggtgc tcctcgttca catacaggtg gcagatacgg cggtacacct cgccctgctc 22560 gggcatcagc tggaaggcgg acttcaggtc gctctccacg cggtaccggt ccatcagcag 22620 cgtcatcact tccatgccct tctcccaggc cgaaacgatc ggcaggctca gggggttctt 22680 caccgttgtc atcttagtcg ccgccgccga agtcaggggg tcgttctcgt ccagggtctc 22740 aaacactcgc ttgccgtcct tctcggtgat gcgcacgggg ggaaagctga agcccacggc 22800 cgccagctcc tcctcggcct gcctttcgtc ctcgctgtcc tggctgatgt cttgcaaagg 22860 cacatgcttg gtcttgcggg gtttcttttt gggcggcaga ggcggcggcg gagacgtgct 22920 gggcgagcgc gagttctcgc tcaccacgac tatttcttct ccttggccgt cgtccgagac 22980 cacgcggcgg taggcatgcc tcttctgggg cagaggcgga ggcgacgggc tctcgcggtt 23040 cggcgggcgg ctggcagagc cccttccgcg ttcgggggtg cgctcctggc ggcgctgctc 23100 tgactgactt cctccgcggc cggccattgt gttctcctag ggagcaagca tggagactca 23160 gccatcgtcg ccaacatcgc catctgcccc cgccgccgcc gacgagaacc agcagcagca 23220 gaatgaaagc ttaaccgccc cgccgcccag ccccacctcc gacgccgcag ccccagacat 23280 gcaagagatg gaggaatcca tcgagattga cctgggctac gtgacgcccg cggagcacga 23340 ggaggagctg gcagcgcgct tttcagcccc ggaagagaac caccaagagc agccagagca 23400 ggaagcagag agcgagcaga accaggctgg gctcgagcat ggcgactacc tgagcggggc 23460 agaggacgtg ctcatcaagc atctggcccg ccaatgcatc atcgtcaagg acgcgctgct 23520 cgaccgcgcc gaggtgcccc tcagcgtggc ggagctcagc cgcgcctacg agcgcaacct 23580 cttctcgccg cgcgtgcccc ccaagcgcca gcccaacggc acctgcgagc ccaacccgcg 23640 cctcaacttc tacccggtct tcgcggtgcc cgaggccctg gccacctacc acctcttttt 23700 caagaaccaa aggatccccg tctcctgccg cgccaaccgc acccgcgccg acgccctgct 23760 caacctgggc cccggcgccc gcctacctga tatcgcctcc ttggaagagg ttcccaagat 23820 cttcgagggt ctgggcagcg acgagactcg ggccgcgaac gctctgcaag gaagcggaga 23880 ggagcatgag caccacagcg ccctggtgga gttggaaggc gacaacgcgc gcctggcggt 23940 cctcaagcgc acggtcgagc tgacccactt cgcctacccg gcgctcaacc tgccccccaa 24000 ggtcatgagc gccgtcatgg accaggtgct catcaagcgc gcctcgcccc tctcggagga 24060 ggagatgcag gaccccgaga gctcggacga gggcaagccc gtggtcagcg acgagcagct 24 120 ggcgcgctgg ctgggagcga gtagcacccc ccagagcctg gaagagcggc gcaagctcat 24180 gatggccgtg gtcctggtga ccgtggagct ggagtgtctg cgccgcttct tcgccgacgc 24240 ggagaccctg cgcaaggtcg aggagaacct gcactacctc ttcagacacg ggttcgtgcg 24300 ccaggcctgc aagatctcca acgtggagct gaccaacctg gtctcctaca tgggcatcct 24360 gcacgagaac cgcctggggc agaacgtgct gcacaccacc ctgcgcgggg aggcccgccg 24420 cgactacatc cgcgactgcg tctacctgta cctctgccac acctggcaga cgggcatggg 24480 cgtgtggcag cagtgcctgg aggagcagaa cctgaaagag ctctgcaagc tcctgcagaa 24540 gaacctcaag gccctgtgga ccgggttcga cgagcgcacc accgccgcgg acctggccga 24600 cctcatcttc cccgagcgcc tgcggctgac gctgcgcaac gggctgcccg actttatgag 24660 ccaaagcatg ttgcaaaact ttcgctcttt catcctcgaa cgctccggga tcctgcccgc 24720 cacctgctcc gcgctgccct cggacttcgt gccgctgacc ttccgcgagt gccccccgcc 24780 gctctggagc cactgctacc tgctgcgcct ggccaactac ctggcctacc actcggacgt 24840 gatcgaggac gtcagcggcg agggcctgct cgagtgccac tgccgctgca acctctgcac 24900 gccgcaccgc tccctggcct gcaaccccca gctgctgagc gagacccaga tcatcggcac 24960 cttcgagttg caaggccccg gcgagggcaa ggggggtctg aaactcaccc cggggctgtg 25020 gacctcggcc tacttgcgca agttcgtgcc cgaggactac catcccttcg agatcaggtt 25080 ctacgaggac caatcccagc cgcccaaggc cgagctgtcg gcctgcgtca tcacccaggg 25140 ggccatcctg gcccaattgc aagccatcca gaaatcccgc caagaatttc tgctgaaaaa 25200 gggccacggg gtctacttgg acccccagac cggagaggag ctcaacccca gcttccccca 25260 ggatgccccg aggaagcagc aagaagctga aagtggagct gccgccgccg ccggaggatt 25320 tggaggaaga ctgggagagc agtcaggcag aggaggagga gatggaagac tgggacagca 25380 ctcaggcaga ggaggacagc ctgcaagaca gtctggagga ggaagacgag gtggaggagg 25440 cagaggaaga agcagccgcc gccagaccgt cgtcctcggc ggaggaggag aaagcaagca 25500 gcacggatac catctccgct ccgggtcggg gtcgcggcgg ccgggcccac agtagatggg 25560 acgagaccgg gcgcttcccg aaccccacca cccagaccgg taagaaggag cggcagggat 25620 acaagtcctg gcgggggcac aaaaacgcca tcgtctcctg cttgcaagcc tgcgggggca 25680 acatctcctt cacccggcgc tacctgctct tccaccgcgg ggtgaacttc ccccgcaaca 25740 tcttgcatta ctaccgtcac ctccacagcc cctactactg tttccaagaa gaggcagaaa 25800 cccagcagca gcagcagcag cagaaaacca gcggcagcag ctagaaaatc cacagcggcg 25860 gcaggtggac tgaggatcgc ggcgaacgag ccggcgcaga cccgggagct gaggaaccgg 25920 atctttccca ccctctatgc catcttccag cagagtcggg ggcaagagca ggaactgaaa 25980 gtcaagaacc gttctctgcg ctcgctcacc cgcagttgtc tgtatcacaa gagcgaagac 26040 caacttcagc gcactctcga ggacgccgag gctctcttca acaagtactg cgcgctcact 26100 cttaaagagt agcccgcgcc cgcccacaca cggaaaaagg cgggaattac gtcaccacct 26160 gcgcccttcg cccgaccatc atcatgagca aagagattcc cacgccttac atgtggagct 26220 accagcccca gatgggcctg gccgccggcg ccgcccagga ctactccacc cgcatgaact 26280 ggctcagtgc cgggcccgcg atgatctcac gggtgaatga catccgcgcc caccgaaacc 26340 agatactcct agaacagtca gcgatcaccg ccacgccccg ccatcacctt aatccgcgta 26400 attggcccgc cgccctggtg taccaggaaa ttccccagcc cacgaccgta ctacttccgc 26460 gagacgccca ggccgaagtc cagctgacta actcaggtgt ccagctggcc ggcggcgccg 26520 ccctgtgtcg tcaccgcccc gctcagggta taaagcggct ggtgatccga ggcagaggca 26580 cacagctcaa cgacgaggtg gtgagctctt cgctgggtct gcgacctgac ggagtcttcc 26640 aactcgccgg atcggggaga tcttccttca cgcctcgtca ggccgtcctg actttggaga 26700 gttcgtcctc gcagccccgc tcgggtggca tcggcactct ccagttcgtg gaggagttca 26760 ctccctcggt ctacttcaac cccttctccg gctcccccgg ccactacccg gacgagttca 26820 tcccgaactt cgacgccatc agcgagtcgg tggacggcta cgattgaatg tcccatggtg 26880 gcgcggctga cctagctcgg cttcgacacc tggaccactg ccgccgcttc cgctgcttcg 26940 ctcgggatct cgccgagttt gcctactttg agctgcccga ggagcaccct cagggcccgg 27000 cccacggagt gcggatcgtc gtcgaagggg gtctcgactc ccacctgctt cggatcttca 27060 gccagcgtcc gatcctggcc gagcgcgagc aaggacagac ccttctgacc ctgtactgca 27120 tctgcaacca ccccggcctg catgaaagtc tttgttgtct gctgtgtact gagtataata 27180 aaagctgaga tcagcgacta ctccggactt ccgtgtgttc ctgctatcaa ccagtccctg 27240 ttcttcaccg ggaacgagac cgagctccag ctccagtgta agccccacaa gaagtacctc 27300 acctggctgt tccagggctc tccgatcgcc gttgtcaacc actgcgacaa cgacggagtc 27360 ctgctgagcg gccctgccaa ccttactttt tccacccgca gaagcaagct ccagctcttc 27420 caacccttcc tccccgggac ctatcagtgc gtctcgggac cctgccatca caccttccac 27480 ctgatcccga ataccacagc gtcgctcccc gctactaaca accaaactac ccaccaacgc 27540 caccgtcgcg acctttcctc tgggtctaat accactaccg gaggtgagct ccgaggtcga 27600 ccaacctctg ggatttacta cggcccctgg gaggtggtag ggttaatagc gctaggccta 27660 gttgcgggtg ggcttttggc tctctgctac ctatacctcc cttgctgttc gtacttagtg 27720 gtgctgtgtt gctggtttaa gaaatgggga agatcaccct agtgagctgc ggtgtgctgg 27780 tggcggtggt gctttcgatt gtgggactgg gcggcgcggc tgtagtgaag gagaaggccg 27840 atccctgctt gcatttcaat cccgacaaat gccagctgag ttttcagccc gatggcaatc 27900 ggtgcgcggt gctgatcaag tgcggatggg aatgcgagaa cgtgagaatc gagtacaata 27960 acaagactcg gaacaatact ctcgcgtccg tgtggcagcc cggggacccc gagtggtaca 28020 ccgtctctgt ccccggtgct gacggctccc cgcgcaccgt gaataatact ttcatttttg 28080 cgcacatgtg cgacacggtc atgtggatga gcaagcagta cgatatgtgg ccccccacga 28140 aggagaacat cgtggtcttc tccatcgctt acagcgtgtg cacggcgcta atcaccgcta 28200 tcgtgtgcct gagcattcac atgctcatcg ctattcgccc cagaaataat gccgaaaaag 28260 aaaaacagcc ataacacgtt ttttcacaca cctttttcag accatggcct ctgttaaatt 28320 tttgctttta tttgccagtc tcattgccgt cattcatgga atgagtaatg agaaaattac 28380 tatttacact ggcactaatc acacattgaa aggtccagaa aaagccacag aagtttcatg 28440 gtattgttat tttaatgaat cagatgtatc tactgaactc tgtggaaaca ataacaaaaa 28500 aaatgagagc attactctca tcaagtttca atgtggatct gacttaaccc taattaacat 28560 cactagagac tatgtaggta tgtattatgg aactacagca ggcatttcgg acatggaatt 28620 ttatcaagtt tctgtgtctg aacccaccac gcctagaatg accacaacca caaaaactac 28680 acctgttacc actatacagc tcactaccaa tggctttctt gccatgcttc aagtggctga 28740 aaatagcacc agcattcaac ccaccccacc cagtgaggaa attcccagat ccatgattgg 28800 cattattgtt gctgtagtgg tgtgcatgtt gatcatcgcc ttgtgcatgg tgtactatgc 28860 cttctgctac agaaagcaca gactgaacga caagctggaa cacttactaa gtgttgaatt 28920 ttaatttttt agaaccatga agatcctagg ccttttagtt ttttctatca ttacctctgc 28980 tctatgcaat tctgacaatg aggacgttac tgtcgttgtc ggatcaaatt atacactaaa 29040 aggtccagca aaaggtatgc tttcgtggta ttgttggttc ggaactgacg agcaacagac 29100 agaactttgc aatgctcaaa aaggcaaaac ctcaaattct aaaatctcta attatcaatg 29160 caatggcact gacttagtat tgctcaatgt cacgaaagca tatgctggca gttacacctg 29220 ccctggagat gatgccgaca atatgatttt ttacaaagtg gaagtggttg atcccactac 29280 tccaccgccc accaccacaa ctactcatac cacacacaca gaacaaacac cagaggcagc 29340 agaagcagag ttggccttcc aggttcacgg agattccttt gctgtcaata cccctacacc 29400 cgatcagcgg tgtccggggc tgctcgtcag cggcattgtc ggtgtgcttt cgggattagc 29460 agtcataatc atctgcatgt tcatttttgc ttgctgctat agaaggcttt accgacaaaa 29520 atcagaccca ctgctgaacc tctatgttta attttttcca gagccatgaa ggcagttagc 29580 gctctagttt tttgttcttt gattggcatt gtttttagtg ctgggttttt gaaaaatctt 29640 accatttatg aaggtgagaa tgccactcta gtgggcatca gtggtcaaaa tgtcagctgg 29700 ctaaaatacc atctagatgg gtggaaagac atttgcgatt ggaatgtcac tgtgtataca 29760 tgtaatggag ttaacctcac cattactaat gccacccaag atcagaatgg taggtttaag 29820 ggccagagtt tcactagaaa taatgggtat gaatcccata acatgtttat ctatgacgtc 29880 actgtcatca gaaatgagac tgccaccacc acacagatgc ccactacaca cagttctacc 29940 actactacca tgcaaaccac acagacaacc actacatcaa ctcagcatat gaccaccact 30000 acagcagcaa agccaagtag tgcagcgcct cagccccagg ctttggcttt gaaagctgca 30060 caacctagta caactactag gaccaatgag cagactactg aatttttgtc cactgtcgag 30120 agccacacca cagctacctc cagtgccttc tctagcaccg ccaatctctc ctcgctttcc 30180 tctacaccaa tcagtcccgc tactactccc accccagctc ttctccccac tcccctgaag 30240 caaactgagg acagcggcat gcaatggcag atcaccctgc tcattgtgat cgggttggtc 30300 atcctggccg tgttgctcta ctacatcttc tgccgccgca ttcccaacgc gcaccgcaaa 30360 ccggcctaca agcccatcgt tatcgggcag ccggagccgc ttcaggtgga agggggtcta 30420 aggaatcttc tcttctcttt tacagtatgg tgattgaact atgattccta gacaattctt 30 480 gatcactatt cttatctgcc tcctccaagt ctgtgccacc ctcgctctgg tggccaacgc 30540 cagtccagac tgtattgggc ccttcgcctc ctacgtgctc tttgccttca tcacctgcat 30600 ctgctgctgt agcatagtct gcctgcttat caccttcttc cagttcattg actggatctt 30660 tgtgcgcatc gcctacctgc gccaccaccc ccagtaccgc gaccagcgag tggcgcggct 30720 gctcaggctc ctctgataag catgcgggct ctgctacttc tcgcgcttct gctgttagtg 30780 ctcccccgcc ccgtcgaccc ccggtccccc actcagtccc ccgaagaggt ccgcaaatgc 30840 aaattccaag aaccctggaa attcctcaaa tgctaccgcc aaaaatcaga catgcttccc 30900 agctggatca tgatcattgg gatcgtgaac attctggcct gcaccctcat ctcctttgtg 30960 atttacccct gctttgactt tggttggaac tcgccagagg cgctctatct cccgcctgaa 31020 cctgacacac caccacagca acctcaggca cacgcactac caccaccaca gcctaggcca 31080 caatacatgc ccatattaga ctatgaggcc gagccacagc gacccatgct ccccgctatt 31140 agttacttca atctaaccgg cggagatgac tgacccactg gccaacaaca acgtcaacga 31200 ccttctcctg gacatggacg gccgcgcctc ggagcagcga ctcgcccaac ttcgcattcg 31260 ccagcagcag gagagagccg tcaaggagct gcaggacggc atagccatcc accagtgcaa 31320 gaaaggcatc ttctgcctgg tgaaacaggc caagatctcc tacgaggtca ccccgaccga 31380 ccatcgcctc tcctacgagc tcctgcagca gcgccagaag ttcacctgcc tggtcggagt 31440 caaccccatc gtcatcaccc agcagtcggg cgataccaag gggtgcatcc actgctcctg 31500 cgactccccc gactgcgtcc acactctgat caagaccctc tgcggcctcc gcgacctcct 31560 ccccatgaac taatcacccc cttatccagt gaaataaata tcatattgat gatgatttaa 31620 ataaaaaata atcatttgat ttgaaataaa gatacaatca tattgatgat ttgagtttta 31680 aaaaataaag aatcacttac ttgaaatctg ataccaggtc tctgtccatg ttttctgcca 31740 acaccacctc actcccctct tcccagctct ggtactgcag accccggcgg gctgcaaact 31800 tcctccacac gctgaagggg atgtcaaatt cctcctgtcc ctcaatcttc attttatctt 31860 ctatcagatg tccaaaaagc gcgtccgggt ggatgatgac ttcgaccccg tctaccccta 31920 cgatgcagac aacgcaccga ccgtgccctt catcaacccc cccttcgtct cttcagatgg 31980 attccaagag aagcccctgg gggtgctgtc cctgcgactg gctgaccccg tcaccaccaa 32040 gaacggggaa atcaccctca agctgggaga gggggtggac ctcgactcct cgggaaaact 32100 catctccaac acggccacca aggccgccgc ccctctcagt ttttccaaca acaccatttc 32160 ccttaacatg gatacccctc tttataccaa agatggaaaa ttatccttac aagtttctcc 32220 accgttaaac atattaaaat caaccattct gaacacatta gctgtagctt atggatcagg 32280 tttaggactg agtggtggca ctgctcttgc agtacagttg gcctctccac tcacttttga 32340 tgaaaaagga aatattaaaa ttaacctagc cagtggtcca ttaacagttg atgcaagtcg 32400 acttagtatc aactgcaaaa gaggggtcac tgtcactacc tcaggagatg caattgaaag 32460 caacataagc tggcctaaag gtataagatt tgaaggtaat ggcatagctg caaacattgg 32520 cagaggattg gaatttggaa ccactagtac agagactgat gtcacagatg catacccaat 32580 tcaagttaaa ttgggtactg gccttacctt tgacagtaca ggcgccattg ttgcttggaa 32640 caaagaggat gataaactta cattatggac cacagccgac ccctcgccaa attgcaaaat 32700 atactctgaa aaagatgcca aactcacact ttgcttgaca aagtgtggaa gtcaaattct 32760 gggtactgtg actgtattgg cagtgaataa tggaagtctc aacccaatca caaacacagt 32820 aagcactgca ctcgtctccc tcaagtttga tgcaagtgga gttttgctaa gcagctccac 32880 attagacaaa gaatattgga acttcagaaa gggagatgtt acacctgctg agccctatac 32940 taatgctata ggttttatgc ctaacataaa ggcctatcct aaaaacacat ctgcagcttc 33000 aaaaagccat attgtcagtc aagtttatct caatggggat gaggccaaac cactgatgct 33060 gattattact tttaatgaaa ctgaggatgc aacttgcacc tacagtatca cttttcaatg 33120 gaaatgggat agtactaagt acacaggtga aacacttgct accagctcct tcaccttctc 33180 ctacatcgcc caagaatgaa cactgtatcc caccctgcat gccaaccctt cccaccccac 33240 tctgtctatg gaaaaaactc tgaagcacaa aataaaataa agttcaagtg ttttattgat 33300 tcaacagttt tacaggattc gagcagttat ttttcctcca ccctcccagg acatggaata 33360 caccaccctc tccccccgca cagccttgaa catctgaatg ccattggtga tggacatgct 33420 tttggtctcc acgttccaca cagtttcaga gcgagccagt ctcgggtcgg tcagggagat 33480 gaaaccctcc gggcactccc gcatctgcac ctcacagctc aacagctgag gattgtcctc 33540 ggtggtcggg atcacggtta tctggaagaa gcagaagagc ggcggtggga atcatagtcc 33600 gcgaacggga tcggccggtg gtgtcgcatc aggccccgca gcagtcgctg ccgccgccgc 33660 tccgtcaagc tgctgctcag ggggtccggg tccagggact ccctcagcat gatgcccacg 33720 gccctcagca tcagtcgtct ggtgcggcgg gcgcagcagc gcatgcggat ctcgctcagg 33780 tcgctgcagt acgtgcaaca caggaccacc aggttgttca acagtccata gttcaacacg 33840 ctccagccga aactcatcgc gggaaggatg ctacccacgt ggccgtcgta ccagatcctc 33900 aggtaaatca agtggcgctc cctccagaac acgctgccca cgtacatgat ctccttgggc 33960 atgtggcggt tcaccacctc ccggtaccac atcaccctct ggttgaacat gcagccccgg 34020 atgatcctgc ggaaccacag ggccagcacc gccccgcccg ccatgcagcg aagagacccc 34080 gggtcccggc aatggcaatg gaggacccac cgctcgtacc cgtggatcat ctgggagctg 34140 aacaagtcta tgttggcaca gcacaggcat atgctcatgc atctcttcag cactctcagc 34200 tcctcggggg tcaaaaccat atcccagggc acggggaact cttgcaggac agcgaacccc 34260 gcagaacagg gcaatcctcg cacataactt acattgtgca tggacagggt atcgcaatca 34320 ggcagcaccg ggtgatcctc caccagagaa gcgcgggtct cggtctcctc acagcgtggt 34380 aagggggccg gccgatacgg gtgatggcgg gacgcggctg atcgtgttcg cgaccgtgtc 34440 atgatgcagt tgctttcgga cattttcgta cttgctgtag cagaacctgg tccgggcgct 34500 gcacaccgat cgccggcggc ggtcccggcg cttggaacgc tcggtgttga aattgtaaaa 34560 cagccactct ctcagaccgt gcagcagatc tagggcctca ggagtgatga agatcccatc 34620 atgcctgata gctctgatca catcgaccac cgtggaatgg gccagaccca gccagatgat 34680 gcaattttgt tgggtttcgg tgacggcggg ggagggaaga acaggaagaa ccatgattaa 34740 cttttaatcc aaacggtctc ggagcacttc aaaatgaagg tcgcggagat ggcacctctc 34800 gcccccgctg tgttggtgga aaataacagc caggtcaaag gtgatacggt tctcgagatg 34860 ttccacggtg gcttccagca aagcctccac gcgcacatcc agaaacaaga caatagcgaa 34920 agcgggaggg ttctctaatt cctcaatcat catgttacac tcctgcacca tccccagata 34980 attttcattt ttccagcctt gaatgattcg aactagttcc tgaggtaaat ccaagccagc 35040 catgataaag agctcgcgca gagcgccctc caccggcatt cttaagcaca ccctcataat 35100 tccaagatat tctgctcctg gttcacctgc agcagattga caagcggaat atcaaaatct 35160 ctgccgcgat ccctaagctc ctccctcagc aataactgta agtactcttt catatcctct 35220 ccgaaatttt tagccatagg accaccagga ataagattag ggcaagccac agtacagata 35280 aaccgaagtc ctccccagtg agcattgcca aatgcaagac tgctataagc atgctggcta 35340 gacccggtga tatcttccag ataactggac agaaaatcac ccaggcaatt tttaagaaaa 35400 tcaacaaaag aaaaatcctc caggtgcacg tttagagcct cgggaacaac gatgaagtaa 35460 atgcaagcgg tgcgttccag catggttagt tagctgatct gtaaaaaaca aaaaataaaa 35520 cattaaacca tgctagcctg gcgaacaggt gggtaaatcg ttctctccag caccaggcag 35580 gccacggggt ctccggcgcg accctcgtaa aaattgtcgc tatgattgaa aaccatcaca 35640 gagagacgtt cccggtggcc ggcgtgaatg attcgacaag atgaatacac ccccggaaca 35700 ttggcgtccg cgagtgaaaa aaagcgcccg aggaagcaat aaggcactac aatgctcagt 35760 ctcaagtcca gcaaagcgat gccatgcgga tgaagcacaa aatcctcagg tgcgtacaaa 35820 atgtaattac tcccctcctg cacaggcagc gaagcccccg atccctccag atacacatac 35880 aaagcctcag cgtccatagc ttaccgagca gcagcacaca acaggcgcaa gagtcagaga 35940 aaggctgagc tctaacctgt ccacccgctc tctgctcaat atatagccca gatctacact 36000 gacgtaaagg ccaaagtcta aaaatacccg ccaaataatc acacacgccc agcacacgcc 36060 cagaaaccgg tgacacactc aaaaaaatac gcgcacttcc tcaaacgccc aaactgccgt 36 120 catttccggg ttcccacgct acgtcatcgg aattcgactt tcaaattccg tcgaccgtta 36180 aaaacgtcac ccgccccgcc cctaacggtc gcccgtctct cggccaatca ccttcctccc 36240 tccccaaatt caaacagctc atttgcatat taacgcgcac caaaagtttg aggtatatta 36300 ttgatgatga tcttaattaa tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa 36360 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 36420 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 36480 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 36540 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 36600 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 36660 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 36720 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 36780 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 36840 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 36900 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 36960 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct 37020 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 37080 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 37140 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 37200 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 37260 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 37320 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 37380 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 37440 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 37500 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 37560 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 37620 gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 37680 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 37740 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 37800 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 37860 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 37920 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 37980 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 38040 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 38100 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 38160 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 38220 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 38280 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta 38340 acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaatttta attaaccc 38398

Claims (26)

Replication deficient simian adenovirus vector C7 encoding a protein comprising a CS protein derived from P. falciparum or a fragment thereof. The vector of claim 1, wherein the encoded protein comprises the sequence of Seq ID No: 7. 7. The vector of claim 1 or 2, wherein the encoded protein comprises the sequence of Seq ID No: 8. The vector of claim 1, wherein the encoded protein has the sequence of Seq ID No: 1. 5. The vector of claim 1 or 2, wherein the encoded protein has the sequence of Seq ID No: 3. A composition comprising a viral vector as defined in any one of claims 1 to 5 and an excipient. A vaccine composition for treating or preventing malaria comprising a viral vector and an adjuvant as defined in any one of claims 1 to 5. 8. The vaccine composition of claim 7, further comprising a protein. The vaccine composition of claim 8, wherein said protein is RTS, S. (1) a vaccine comprising a replication defective Simian Adenovirus Vector C7, (2) malaria antigen, and (3) adjuvant, which encodes a protein comprising a CS protein derived from P. falciparum or a fragment thereof Composition. Kit comprising (1) a replication defective Simian adenovirus vector C7, (2) malaria antigen, and (3) adjuvant, encoding a protein comprising a CS protein derived from P. falciparum or a fragment thereof . The vaccine or kit of claim 10 or 11, wherein the protein encoded by the adenovirus vector C7 has the sequence of Seq ID No: 1. 12. The vaccine or kit according to any one of claims 7 to 12, wherein the malaria antigen is RTS, S. The vaccine or kit according to any one of claims 7 to 13, wherein the adjuvant comprises 3D-MPL. The vaccine or kit of any one of claims 7-13, wherein the adjuvant comprises saponin. The vaccine or kit of claim 15 wherein the saponin is QS21. The vaccine or kit of any of claims 7-13, wherein the adjuvant comprises 3D-MPL and QS21. 18. The vaccine or kit of any one of claims 7 to 17, wherein the formulation is provided in an oil-in-water emulsion. 18. The vaccine or kit of any of claims 7-10 or 12-17, wherein the vaccine is a liposome formulation. The replication defective Simian adenovirus vector C7 according to claim 10, which encodes a protein comprising a CS protein derived from P. falciparum having the sequence of Seq ID No: 1, (2) malaria A vaccine composition comprising an antigen RTS, S, and an adjuvant comprising (3) 3D-MPL and QS21. a. Propagating the vector in a suitable cell line, and
b. Recovering the vector;
A method for producing a viral vector of any one of claims 1 to 5. The composition of claim 6 or any one of claims 7 to 10 or 12 to 20, comprising mixing the viral vector of claim 1 with at least one excipient / carrier. According to the method of preparation of the vaccine. The viral vector according to any one of claims 1 to 5 for the treatment or prevention of malaria. The viral vector as defined in any one of claims 1 to 5 for priming or boosting in a prime boost regimen for the treatment or prevention of malaria. Use of a viral vector as defined in any one of claims 1 to 5 in the manufacture of a medicament for the treatment or prevention of malaria. a. Viral vector as defined in any one of claims 1 to 5,
b. A composition as defined in claim 6, or
c. A method of treatment comprising administering a therapeutically effective amount of a vaccine as defined in any of claims 7 to 10 or 12 to 20.

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