KR20160125155A - A new marker for diagnosis of macular degeneration and a diagnostic method using the smae - Google Patents

Abstract

The present invention relates to a novel macular degeneration marker and a method for diagnosing macular degeneration. More particularly, the present invention relates to a composition for diagnosing macular degeneration associated with a mutation of CFH, CFB, C3 and CFI genes, a diagnostic kit and a sanger sequencing, Such as pyrosequencing and HRM assays, which are capable of rapidly and accurately analyzing such gene mutations. The composition for the diagnosis of macular degeneration, the diagnostic kit and the diagnostic method according to the present invention are novel markers capable of effectively diagnosing macular degeneration, and are excellent in sensitivity, can be quickly and easily analyzed, . ≪ / RTI >

Description

[0001] The present invention relates to a novel marker for macular degeneration and a method for diagnosing macular degeneration,

The present invention relates to a novel macular degeneration marker and a method for diagnosing macular degeneration, and more particularly, to a macular degeneration diagnostic composition and diagnostic marker relating to a mutation of CFH, CFB, C3, CFI gene, sanger sequencing, Such as pyrosequencing and HRM assays, which are capable of rapidly and accurately analyzing such gene mutations.

The nerve tissue located in the center of the inner retina of the eye is called the macula. Most of the photoreactive cells responding to the light stimuli are gathered here, and the phase of the object is also the center of the macula, and plays a very important role in sight. Age-related macular degeneration (AMD) is a chronic disease characterized by macular degeneration of the retinal pigment epithelium, Bruch's membrane, and choroidal capillaries. Anatomically, the sensory retina is located in the anterior part of the retinal pigment epithelium, and the treatment of nutrition, support, recirculation and waste products depends on the retinal pigment epithelium. Bruch's membrane is a five-layered structure that is interposed between the choroid and the retinal pigment epithelium. The innermost layer is the basement membrane of the retinal pigment epithelium and the outermost layer is the basement membrane of the choroidal capillary endothelial cells. In other words, it is a metamorphosis of the retinal pigment epithelium, Bruch's membrane and choroidal capillary complex.

This disease occurs mainly in the age group of 50 years or older. In the West, it is the main cause of blindness in the population over 60 years of age and it is increasing in Korea. The causes of age-related macular degeneration have not yet been elucidated. However, the most known risk factors are age (especially after 75 years of age), high-blood pressure, obesity, genetic predisposition, Exposure, and low serum antioxidant concentrations.

There are two types of macular degeneration: dry (non-exudative) macular degeneration and habitual (exudative) macular degeneration. Dry AMD, nonexudative AMD, and nonneovascular AMD, build up a yellow precipitate called drusen underneath the retina. When it grows larger, it blocks blood flow to the retina, especially the macula, The obstacle begins to come to. Dry AMD does not cause rapid visual loss, but it can progress to hatching macular degeneration. Below the retina is a choroid that contains a collection of blood vessels embedded within the fibrous tissue and a pigment epithelium that covers the choroid layer. Wet AMD (exudative AMD, neovascular AMD) is caused by the growth of new blood vessels in the choroid below the retina. These weak neovasions are bleeding and bleeding and exudation, causing macular degeneration in the retina, resulting in visual disturbances. It is known that macular degeneration progresses very quickly, so the visual acuity rapidly deteriorates within weeks and blindness may occur between two months and three years.

It is known that 70% of progressive macular degeneration is determined by genetic characteristics (CFH, ARMS2, etc.) and the remaining 30% is determined by environmental factors such as smoking and habit. (Nat Genet. 2013 Apr; 45 (4): 433-9, Nat Genet. 2013 Nov; 45 (11) ): 1371-4). However, these gene mutations have been studied in western countries, and studies on Asians (Korean, Japanese, Chinese, etc.) have not yet been conducted. Thus, there is a growing need to exploit additional polymorphic identifications in Far Eastern Asian populations that can be associated with or predict the safety of macular degenerative diseases.

Next Generation Sequencing (NGS) is a technology to read all DNA sequences. As Solexa was merged into Illumina in 2007, the term "Next Generation Sequencing (NGS)" began to be used. . With the development of NGS technology, it becomes possible to analyze large-scale sequence information more easily and inexpensively than traditional methods.

DNA sequence information analyzed through this next generation sequencing can be used to identify individual differences and ethnic characteristics, to identify congenital causes including chromosomal abnormalities in diseases associated with gene abnormalities, and to identify genes that inhibit complex diseases such as macular degeneration, diabetes, and hypertension Defects can be easily found, and information such as gene expression, genetic diversity, and their interactions can be widely used in the field of molecular diagnosis and therapy.

Therefore, the present inventors analyzed a set of related genes for macular degeneration diseases through next generation sequencing, and found that 8 novel mutation genes ( CFH Y56C, CFH L77S, CFH D798E, CFH F176L, CFB G208S, CFB V716L, C3 R1134Q, CFI M204R) and three disease gene mutations ( CFH G1110A, C3 A636S, C3 H16Q), and 11 genes were excavated. These mutations were confirmed by gene analysis (HRM, Taq-man assays).

1. Nat Genet. 2013 Apr; 45 (4): 433-9, Nat Genet. 2013 Nov; 45 (11): 1371-4

It is therefore an object of the present invention to provide a composition for the diagnosis of macular degeneration that can predict the risk of developing macular degeneration and estimate its prognosis and treatment progress.

Another object of the present invention is to provide a kit for the diagnosis of macular degeneration comprising the diagnostic composition.

It is yet another object of the present invention to provide a method for detecting a mutation of a gene associated with a risk of macular degeneration for rapid and accurate analysis of macular degeneration.

It is yet another object of the present invention to provide a macroreticular-specific mutant polypeptide.

In order to achieve the above object, the present invention provides a composition for the detection of macular degeneration, which comprises a preparation capable of detecting a mutation composed of one or two or more combinations selected from a mutation group of the following proteins.

The CFH proteins Y56C, L77S, D798E, F176L, G1110A;

The C3 protein R1134Q, A636S, C3 H16Q;

G208S, V716L of CFB protein; And

M204R of CFI protein.

In one embodiment of the present invention, the agent may be a primer pair or a probe that specifically binds to a gene encoding the protein.

In one embodiment of the present invention, the primer pair comprises a sequence selected from the group consisting of SEQ ID NOs: 1 and 2, SEQ ID NOs: 3 and 4, SEQ ID NOs: 5 and 6, SEQ ID NOs: 7 and 8, SEQ ID NOs: 9 and 10, SEQ ID NOs: SEQ ID NOs: 15 and 16, SEQ ID NOs: 17 and 18, SEQ ID NOs: 19 and 20, and combinations of one or more primer pairs selected from the group consisting of SEQ ID NOs:

In one embodiment of the invention, the agent may be an antibody specific for the protein.

On the other hand, the present invention provides a kit for the diagnosis of macular degeneration comprising the above composition.

In one embodiment of the present invention, the kit may be an RT-PCR kit, a microarray chip kit, or a protein chip kit.

In one embodiment of the present invention, the RT-PCR kit may include a primer pair that specifically binds to a gene encoding the protein.

In one embodiment of the present invention, the primer pair comprises a sequence selected from the group consisting of SEQ ID NOs: 1 and 2, SEQ ID NOs: 3 and 4, SEQ ID NOs: 5 and 6, SEQ ID NOs: 7 and 8, SEQ ID NOs: 9 and 10, SEQ ID NOs: SEQ ID NOs: 15 and 16, SEQ ID NOs: 17 and 18, SEQ ID NOs: 19 and 20, and combinations of one or more primer pairs selected from the group consisting of SEQ ID NOs:

In one embodiment of the present invention, the microarray chip kit may include a probe that specifically binds to a gene encoding the protein.

In one embodiment of the present invention, the protein chip kit may comprise an antibody specific for the protein.

The present invention also relates to a method of amplifying DNA comprising the steps of: (i) performing PCR using the primer pairs of SEQ ID NOS: 1 and 2; (ii) pyrosequencing using the sequencing primers of SEQ ID NOS: 3 and 4 for the PCR products obtained in step (i); and detecting the mutation of the gene associated with the risk of macular degeneration .

In one embodiment of the present invention, the gene associated with the risk for macular degeneration may be a complement factor H (CFH).

In one embodiment of the present invention, the amino acid mutation by the CFH gene mutation may be CFH Y56C or L77S.

The present invention also provides a mutant wherein amino acid 56 of SEQ ID NO: 21 is substituted with Tyr to Cys; A mutation in which amino acid 77 is substituted with Leu to Ser; A mutation in which amino acid 798 is substituted with Asp to Glu; A mutation in which amino acid 176 is substituted from Phe to Leu; A mutation in which amino acid 1110 is replaced by Ala in Gly;

A mutation in which amino acid 208 of SEQ ID NO: 22 is substituted with Gly to Ser; A mutation in which amino acid 716 is substituted by Val to Leu;

A mutation in which amino acid 1134 of SEQ ID NO: 23 is substituted from Arg to Gln; A mutant in which amino acid 636 is substituted with Ala to Ser, a mutant in which amino acid 16 is substituted from His to Gln; And

A mutation in which the amino acid at position 204 of SEQ ID NO: 24 is substituted with Arg to Met; and a mutant in which the amino acid at position 204 of SEQ ID NO: 24 is substituted with Arg to Met.

The composition for the diagnosis of macular degeneration, the diagnostic kit and the diagnostic method according to the present invention are novel markers capable of effectively diagnosing macular degeneration, and are excellent in sensitivity, can be quickly and easily analyzed, . ≪ / RTI >

1 shows the results of Sanger sequencing analysis of CFH Y56C, CFH L77S, CFH D798E, CFH G1110A, CFH F176L, C3 H16Q, C3 A636S, C3 R1134Q, CFB G208S, CFB V716L, CFI M204R, CFB G208S, CFB V716L and CFI M204R .
Fig. 2 is a cross- 56YY and CFH 77LL type multiplexed pirosequencing analysis results.
Fig. 3 CFH 56YY, and CFH 77LS multiplexed pi-sequencing analysis results.
4 CFH 56YC, and CFH 77LL multiplexed pi-sequencing analysis results.
FIG. 5 shows results of HRM assay of Aligned Melt Curve and Difference Plot of CFH 1110 GG type (red color) and CFH 1110 GA type (blue color).
FIG. 6 shows results of HRM assay of Aligned Melt Curve and Difference Plot of CFH 176 FF type (red color) and CFH 176FL type (blue color).
FIG. 7 shows the results of HRM assay of Aligned Melt Curve and Difference Plot of C3 16HH type (red color) and C3 16HQ type (blue color).
8 is C3 Aligned Melt Curve of 636AA type (red color) and C3 636AS type (blue color) and Difference Plot were analyzed by HRM assay.
9 is C3 1134RR type (red), C3 1134RQ type (blue) Aligned Melt Curve and Difference Plot were analyzed by HRM assay.
10 is a CFB 208GG type (red), CFB It is the result of analyzing 208GS type (blue) Aligned Melt Curve and Difference Plot by HRM assay.
FIG. 11 shows the result of HRM assay of Aligned Melt Curve and Difference Plot of CFB 716VV type (red color) and CFB 716VL type (blue color).
Fig. 12 is a cross- 204MM type (red), CFI 204MR type (blue) Aligned Melt Curve and Difference Plot by HRM assay.

The definitions of the terms used in the present invention are as follows.

'Genetic polymorphism' refers to a case in which a genetic variation occurs in at least 1% of the population. The insertion, deletion, or substitution of a single nucleotide in DNA is called single nucleotide polymorphism (SNP). Nonsynonymous SNP refers to a change in the base sequence due to SNP, and silent SNP or synonymous SNP does not change the amino acid

The term " polymorphism " refers to a sequence in a sequence of genes that varies within a cluster. Polymorphisms consist of different "alleles ". The arrangement of this polymorphism can be confirmed by its position in the gene and the different amino acids or bases found therein. For example, Y402H CFH indicates that the amino acid position 402 in the CFH gene changes between tyrosine (Y) and histidine (H). These amino acid variations are the result of two possible mutant bases, C and T, which are two different alleles. Since the genotype is composed of two different distinct alleles, any of the various possible variants can be observed in any individual (e. G., CC, CT or TT in this example). The individual polymorphisms are also known to those skilled in the art and are used in, for example, the Single Nucleotide Polymorphism Database (dbSNP) of the Nucleotide Sequence Variation of the nucleotide base mutations available on the NCBI website. ("Reference SNP", "refSNP", or "rs #").

The term " genotype " refers to a specific allele of a particular gene in a cell or tissue sample. In this example, CC, CT or TT are the possible genotypes in the Y402H CFH polymorphism

A "marker" refers to a nucleotide sequence or a coding product thereof (eg, a protein) used as a reference point when identifying a locus or associated locus. The marker may be derived from a genomic nucleotide sequence or from a nucleotide sequence expressed (e.g., from RNA, nRNA, mRNA, cDNA, etc.), or from a coded polypeptide. The term includes nucleic acid sequences that are complementary to or flanked in the marker sequence, such as a probe or a pair of primers that can amplify the marker sequence.

"Nucleic acid" refers to polynucleotides or oligonucleotides such as deoxyribonucleic acid (DNA), and, if appropriate, ribonucleic acid (RNA). The term also encompasses both single (sense or antisense) and double helical polynucleotides, as applied to the described embodiments, as well as analogs of either RNA or DNA prepared from the nucleotide analogs (e.g., peptide nucleic acids) . In the present invention, the term 'nucleic acid' and the term 'nucleotide' are used in combination.

'Primer' refers to an oligonucleotide sequence that hybridizes to complementary RNA or DNA-targeted polynucleotides and serves as a starting point for the stepwise synthesis of polynucleotides from mononucleotides, for example by the action of a nucleotidyltransferase that occurs in the polymerase chain reaction .

&Quot; Diseases and conditions associated with polymorphism " refers to a susceptibility that can be expressed in a subject based on a variety of diseases or conditions, i.e., the identification of one or more alleles.

'Risk' refers to a statistically high incidence of a disease or condition in a subject having a particular polymorphic allele compared to the incidence of a disease or condition in a member of the individual that does not possess the polymorphic allele.

The 'phenotype' is the phenotypic, physiological, or biochemical characteristic of an individual determined by the genotype. Genotypes are distinct from phenotypes, which refer to individual gene constructs and, in a narrower sense, alleles on a gene.

"Target" or "patient" means any single entity requiring treatment, including human, cow, dog, guinea pig, rabbit, chicken, insect, and the like. In addition, any subject who participates in a clinical study test that does not show any disease clinical findings, or who participates in epidemiological studies or used as a control group is included. In one embodiment of the present invention, the present invention was applied to humans.

&Quot; Tissue or cell sample " refers to a collection of similar cells from a subject or tissue of a patient. The source of the tissue or cell sample may be a solid tissue from fresh, frozen and / or preserved organ or tissue sample or biopsy or aspirate; Blood or any blood components; It may be a cell at any point in the pregnancy or development of the subject. Tissue samples can also be primary or cultured cells or cell lines.

An " effective amount " is an appropriate amount that affects a beneficial or desired clinical or biochemical outcome. An effective amount may be administered one or more times. For purposes of the present invention, an effective amount of an inhibitor compound is an amount sufficient to temporarily alleviate, ameliorate, stabilize, reverse, slow down, or delay the progression of a disease state. If the recipient animal is capable of enduring the administration of the composition, or the administration of the composition to the animal is suitable, the composition will be "pharmaceutically or physiologically acceptable ". If the dose administered is physiologically significant, it can be said that the formulation is administered in a "therapeutically effective amount ". The formulation is physiologically relevant if the presence of the formulation results in a physiologically detectable change in the recipient.

The term "treating", unless otherwise indicated, refers to reversing, alleviating, inhibiting, or preventing the disease or condition to which the term applies, or one or more symptoms of the disease or disorder . As used herein, the term " treatment " refers to an act of treating when " treating " is defined as above.

All technical terms used in the present invention are used in the sense that they are generally understood by those of ordinary skill in the relevant field of the present invention unless otherwise defined. Also, preferred methods or samples are described in this specification, but similar or equivalent ones are also included in the scope of the present invention. The contents of all publications referred to herein are incorporated herein by reference.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Unless otherwise indicated, nucleic acids are written in 5'3 'orientation from left to right. The numerical ranges recited in the specification include numerals defining the ranges and include each integer or any non-integral fraction within a defined range.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice of testing the present invention, the preferred materials and methods are described herein.

The HRM assay described in the present invention is one of the real time PCR experimental techniques, and genotyping analysis using a small amount of genomic DNA is possible using the difference in Tm values. The HRM assay is an inexpensive assay, primer design is simpler than the probe, and the analysis time is shortened. The sample can be used for a large amount of screening. The dsDNA intercalating dyes used in the HRM assay may be SYBR® GREEN, SYTO9®, LCGreen®, EvaGreen, or the like, preferably SYTO9®.

The HEM assay can be used as a post-PCR assay to determine if there is a genetic mutation in dsDNA after completion of PCR using PCR and other assays. That is, it can be used for the purpose of discrimination of gene mutation, not DNA amplification. When a fluorescent substance that binds to dsDNA is added during PCR reaction, fluorescence detection is possible in the PCR process. After the restriction enzyme treatment, electrophoresis or Sanger sequencing It is possible to confirm the presence of gene mutation within a short time without any process.

Next Generation Sequencing is a DNA sequencing method that analyzes individual genomes to discover variants. It simultaneously processes sequencing reactions in enormous quantities, so you can get information on more than 3 billion genomes in a short period of time. The present inventors performed a next generation sequencing analysis on 298 patients with macular degeneration and 188 controls, and found a novel mutation 11 that is expressed only in Huqa degeneration.

Example  1: Next Generation Sequencing ( NGS ) To discover mutant genes

Blood samples were drawn from subjects in the disease group and the control group, and DNA was isolated using Qiagen's genomic DNA isolation kit.

DNA sequencing was performed with the Illumina HiSeq sequencing instrument. As a result of the analysis, only eight mutation genes ( CFH Y56C, CFH L77S, CFH D798E, CFH F176L, CFB G208S, CFB V716L, C3 R1134Q, CFI M204R) and three mutant genes ( CFH G1110A, C3 A636S, C3 H16Q). These genes confirmed the authenticity of gene displacement through sanger sequencing.

Next-Generation Sequencing-Targeted NGS is a method of sequencing a specific region of a gene or a genome region. By capturing a specific kit capable of capturing a specific region, it is amplified and then sequenced to obtain raw data. Analyze.

CFH has an average coverage of over 100X, coverage over 100X is 95.4%, coverage of over 100X is more than 20X, coverage of over 100X is average 95.3% The average of 98.1% and the coverage over 100X was 62.7% The results of the next generation sequencing average coverage according to the depth of each gene are shown in Table 1 below, and the results of confirming sanger sequencing are shown in FIG.

gene
Coverage 1X 5X 10X 20X 30X 40X 50X 100X CFH 100.0 100.0 99.9 99.7 99.4 99.0 98.4 94.3 CFB 100.0 100.0 100.0 100.0 100.0 99.9 99.7 95.4 C3 100.0 100.0 99.8 98.1 94.2 89.9 85.6 62.7

Example  2: High-speed mutation genotype analysis of novel mutant genes

11 mutations in the disease group ( CFH Y56C, CFH L77S, CFH D798E, CFH G1110A, CFH F176L, CFB G208S, CFB V716L, CFI M204R, C3 R1134Q, C3 A636S, C3 H16Q) was confirmed by using an automatic sequencer and a next generation sequencing analyzer. On the other hand, we developed a rapid mutation genotyping method for major mutation SNPs for disease group prediction. In detail, multiplex-pyrosequencing of two CFH genes - CFH Y56C and CFH L77S - Two CFH mutation genes ( CFH F176L, CFH G1110A), two CFB species ( CFB G208S, CFB V716L), three C3 mutation genes ( C3 H16Q, C3 A636S, C3 R1134Q), CFI HR20 assay was performed on M204R.

Example  2-1: multiplex - pyrosequencing High-speed screening of mutant genes using

Table 2 below shows the set of primers used for PCR.

Primer name Primer sequences (5 'to 3') SEQ ID NO: Size (bp) CFH_Y56C / L77S_F TCC AGA AGG CAC CCA GGC TA SEQ ID NO: 1 338
CFH_Y56C / L77S_BR * CCA GGC GAT AGA GGG AGA CT SEQ ID NO: 2

(*: It is made by attaching Biotin to 5 'end.)

Table 3 below shows PCR reaction conditions.

PCR 제품 Reaction conditions CFH_Y56C / L778 35 cycles of 94 ° C for 5 minutes, (94 ° C for 30 seconds, 60 ° C for 30 seconds, and 72 ° C for 30 seconds), 72 ° C for 5 minutes

Table 4 below shows primers for sequencing.

Primer name Primer sequences (5 'to 3') SEQ ID NO: CFH_Y56C_SF CTA TAA ATG CCG CCC TGG AT SEQ ID NO: 3 CFH_L77S_SF AGA ATG GGT TGC TCT TAA TC SEQ ID NO: 4

Example  2-2: HRM assay High-speed screening of mutant genes using

The HRM assay is one of the real-time PCR techniques. Genotyping genotyping can be performed using only a small amount of genomic DNA (100 pg to 10 ng) using the difference in Tm value. In the present invention, HRM assay is performed according to the following primer and PCR conditions .

Table 5 below shows the set of primers used for PCR.

Primer name Primer sequences (5 'to 3') SEQ ID NO: Size (bp) CFH_G1110A_F AGT GCT GTG TTT GCG TTT GC SEQ ID NO: 5 178
CFH_G1110A_R CAA CTG ATG AAG CTG GAGCA SEQ ID NO: 6 CFH_F176L_F CCA GTG ACA GCA CCA GAG AA SEQ ID NO: 7 156
CFH_F176L _R ACT CCA AAA ACC ATC GTC TGA SEQ ID NO: 8 CFB_G208S_F TTG CTC TCT ACC TTG CTC ACG SEQ ID NO: 9 309
CFB_G208S_R GAG GTG GGG AGA GAA GAC AGT SEQ ID NO: 10 CFB_V716L_F TGT CTT GCC TGC GTG TGTCA SEQ ID NO: 11 234
CFB_V716L_R TGT GAA AGT CTC GGG CGT GA SEQ ID NO: 12 C3_H16Q_F GCT GCT CCT GCT ACT AAC CC SEQ ID NO: 13 187
C3_H16Q_R CCA AAT GTC TGC TTC CAC CC SEQ ID NO: 14 C3_A636S _F GGT GGA GAA GGC AGA CAT CG SEQ ID NO: 15 95
C3_A636S _R CAC TGC TGC TCG TGA AGG TCA SEQ ID NO: 16 C3_R1134Q_F CCA TCC TTG CTT TCT GCT CT SEQ ID NO: 17 173
C3_R1134Q_R GCA TCG GGT AAG GTA GGG TA SEQ ID NO: 18 CFI_M204R_F CAT TGC CGA GGA TTA GAG ACC SEQ ID NO: 19 116
CFI_M204R_R CTC CAC CAA CCT GCT TTC TG SEQ ID NO: 20

Table 6 below shows the reactant conditions containing fluorescent dye for HRM analysis.

Furtherance volume( ul / sample ) 2X buffer 10 MeltDoctor ™ One Forward primer (5 pmol / ul) One Reverse primer (5 pmol / ul) One genomic DNA (20 ng / ul) 2 ultra pure D.W 5 Sum 20

Table 7 below shows PCR reaction conditions.

PCR 제품 Reaction conditions CFH_G1110A 95 ° C for 10 minutes, (95 ° C for 15 seconds, 60 ° C for 15 seconds, and 72 ° C for 15 seconds) 35 cycles, 95 ° C for 10 seconds, 60 ° C for 1 minute, 95 ° C for 15 seconds, CFH 176L 95 ° C for 10 minutes, (95 ° C for 15 seconds, 60 ° C for 15 seconds, and 72 ° C for 15 seconds) 35 cycles, 95 ° C for 10 seconds, 60 ° C for 1 minute, 95 ° C for 15 seconds, CFB_G208S 95 ° C for 10 minutes, (95 ° C for 15 seconds, 60 ° C for 15 seconds, and 72 ° C for 15 seconds) 35 cycles, 95 ° C for 10 seconds, 60 ° C for 1 minute, 95 ° C for 15 seconds, CFB_V716L 95 ° C for 10 minutes, (95 ° C for 15 seconds, 60 ° C for 15 seconds, and 72 ° C for 15 seconds) 35 cycles, 95 ° C for 10 seconds, 60 ° C for 1 minute, 95 ° C for 15 seconds, C3_H16Q 95 ° C for 10 minutes, (95 ° C for 15 seconds, 60 ° C for 15 seconds, and 72 ° C for 15 seconds) 35 cycles, 95 ° C for 10 seconds, 60 ° C for 1 minute, 95 ° C for 15 seconds, C3_A636S 95 ° C for 10 minutes, (95 ° C for 15 seconds, 60 ° C for 15 seconds, and 72 ° C for 15 seconds) 35 cycles, 95 ° C for 10 seconds, 60 ° C for 1 minute, 95 ° C for 15 seconds, C3_R1134Q 95 ° C for 10 minutes, (95 ° C for 15 seconds, 60 ° C for 15 seconds, and 72 ° C for 15 seconds) 35 cycles, 95 ° C for 10 seconds, 60 ° C for 1 minute, 95 ° C for 15 seconds, CFI_M204R 95 ° C for 10 minutes, (95 ° C for 15 seconds, 60 ° C for 15 seconds, and 72 ° C for 15 seconds) 35 cycles, 95 ° C for 10 seconds, 60 ° C for 1 minute, 95 ° C for 15 seconds,

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

<110> inje university industry-academic cooperation foundation <120> A new marker for diagnosis of macular degeneration and a          diagnostic method using the same <130> PN1501-031 <160> 28 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFH Y56C / L77S PCR forward primer <400> 1 tccagaaggc acccaggcta 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFH Y56C / L77S PCR reverse primer <400> 2 ccaggcgata gagggagact 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFH Y56C sequencing primer <400> 3 ctataaatgc cgccctggat 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFH L77S sequencing primer <400> 4 agaatgggtt gctcttaatc 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFH G1110A PCR forward primer <400> 5 agtgctgtgt ttgcgtttgc 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFH G1110A PCR reverse primer <400> 6 caactgatga agctggagca 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFH F176L PCR forward primer <400> 7 ccagtgacag caccagagaa 20 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CFH F176L PCR reverse primer <400> 8 actccaaaaa ccatcgtctg a 21 <210> 9 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CFB G208S PCR forward primer <400> 9 ttgctctcta ccttgctcac g 21 <210> 10 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CFB G208S PCR reverse primer <400> 10 gaggtgggga gagaagacag t 21 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFB V716L PCR forward primer <400> 11 tgtcttgcct gcgtgtgtca 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFB V716L PCR reverse primer <400> 12 tgtgaaagtc tcgggcgtga 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> &Lt; 223 > C3 H16Q PCR forward primer <400> 13 gctgctcctg ctactaaccc 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> &Lt; 223 > C3 H16Q PCR reverse primer <400> 14 ccaaatgtct gcttccaccc 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> &Lt; 223 > C3 A636S PCR forward primer <400> 15 ggtggagaag gcagacatcg 20 <210> 16 <211> 21 <212> DNA <213> Artificial Sequence <220> &Lt; 223 > C3 A636S PCR reverse primer <400> 16 cactgctgct cgtgaaggtc a 21 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> &Lt; 223 > C3 R1134Q PCR forward primer <400> 17 ccatccttgc tttctgctct 20 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> &Lt; 223 > C3 R1134Q PCR reverse primer <400> 18 gcatcgggta aggtagggta 20 <210> 19 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CFI M204R PCR forward primer <400> 19 cattgccgag gattagagac c 21 <210> 20 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CFI M204R PCR reverse primer <400> 20 ctccaccaac ctgctttctg 20 <210> 21 <211> 1231 <212> PRT <213> Homo sapiens CFH amino acid sequence <400> 21 Met Arg Leu Leu Ala Lys Ile Ile Cys Leu Met Leu Trp Ala Ile Cys   1 5 10 15 Val Ala Glu Asp Cys Asn Glu Leu Pro Pro Arg Arg Asn Thr Glu Ile              20 25 30 Leu Thr Gly Ser Trp Ser Asp Gln Thr Tyr Pro Glu Gly Thr Gln Ala          35 40 45 Ile Tyr Lys Cys Arg Pro Gly Tyr Arg Ser Leu Gly Asn Val Ile Met      50 55 60 Val Cys Arg Lys Gly Glu Trp Val Ala Leu Asn Pro Leu Arg Lys Cys  65 70 75 80 Gln Lys Arg Pro Cys Gly His Pro Gly Asp Thr Pro Phe Gly Thr Phe                  85 90 95 Thr Leu Thr Gly Gly Asn Val Tyr             100 105 110 Thr Cys Asn Glu Gly Tyr Gln Leu Leu Gly Glu Ile Asn Tyr Arg Glu         115 120 125 Cys Asp Thr Asp Gly Trp Thr Asn Asp Ile Pro Ile Cys Glu Val Val     130 135 140 Lys Cys Leu Pro Val Thr Ala Pro Glu Asn Gly Lys Ile Val Ser Ser 145 150 155 160 Ala Met Glu Pro Asp Arg Glu Tyr His Phe Gly Gln Ala Val Arg Phe                 165 170 175 Val Cys Asn Ser Gly Tyr Lys Ile Glu Gly Asp Glu Glu Met His Cys             180 185 190 Ser Asp Gly Phe Trp Ser Lys Glu Lys Pro Lys Cys Val Glu Ile         195 200 205 Ser Cys Lys Ser Pro Asp Val Ile Asn Gly Ser Pro Ile Ser Gln Lys     210 215 220 Ile Ile Tyr Lys Glu Asn Glu Arg Phe Gln Tyr Lys Cys Asn Met Gly 225 230 235 240 Tyr Glu Tyr Ser Glu Arg Gly Asp Ala Val Cys Thr Glu Ser Gly Trp                 245 250 255 Arg Pro Leu Pro Ser Cys Glu Glu Lys Ser Cys Asp Asn Pro Tyr Ile             260 265 270 Pro Asn Gly Asp Tyr Ser Pro Leu Arg Ile Lys His Arg Thr Gly Asp         275 280 285 Glu Ile Thr Tyr Gln Cys Arg Asn Gly Phe Tyr Pro Ala Thr Arg Gly     290 295 300 Asn Thr Ala Lys Cys Thr Ser Thr Gly Trp Ile Pro Ala Pro Arg Cys 305 310 315 320 Thr Leu Lys Pro Cys Asp Tyr Pro Asp Ile Lys His Gly Gly Leu Tyr                 325 330 335 His Glu Asn Met Arg Arg Pro Tyr Phe Pro Val Ala Val Gly Lys Tyr             340 345 350 Tyr Ser Tyr Tyr Cys Asp Glu His Phe Glu Thr Pro Ser Gly Ser Tyr         355 360 365 Trp Asp His Ile His Cys Thr Gln Asp Gly Trp Ser Pro Ala Val Pro     370 375 380 Cys Leu Arg Lys Cys Tyr Phe Pro Tyr Leu Glu Asn Gly Tyr Asn Gln 385 390 395 400 Asn His Gly Arg Lys Phe Val Gln Gly Lys Ser Ile Asp Val Ala Cys                 405 410 415 His Pro Gly Tyr Ala Leu Pro Lys Ala Gln Thr Thr Val Thr Cys Met             420 425 430 Glu Asn Gly Trp Ser Pro Thr Pro Arg Cys Ile Arg Val Lys Thr Cys         435 440 445 Ser Lys Ser Ser Ile Asp Ile Glu Asn Gly Phe Ile Ser Ser Ser Ser Gln     450 455 460 Tyr Thr Tyr Ala Leu Lys Glu Lys Ala Lys Tyr Gln Cys Lys Leu Gly 465 470 475 480 Tyr Val Thr Ala Asp Gly Glu Thr Ser Gly Ser Ile Thr Cys Gly Lys                 485 490 495 Asp Gly Trp Ser Ala Gln Pro Thr Cys Ile Lys Ser Cys Asp Ile Pro             500 505 510 Val Phe Met Asn Ala Arg Thr Lys Asn Asp Phe Thr Trp Phe Lys Leu         515 520 525 Asn Asp Thr Leu Asp Tyr Glu Cys His Asp Gly Tyr Glu Ser Asn Thr     530 535 540 Gly Ser Thr Thr Gly Ser Ile Val Cys Gly Tyr Asn Gly Trp Ser Asp 545 550 555 560 Leu Pro Ile Cys Tyr Glu Arg Glu Cys Glu Leu Pro Lys Ile Asp Val                 565 570 575 His Leu Val Pro Asp Arg Lys Lys Asp Gln Tyr Lys Val Gly Glu Val             580 585 590 Leu Lys Phe Ser Cys Lys Pro Gly Phe Thr Ile Val Gly Pro Asn Ser         595 600 605 Val Gln Cys Tyr His Phe Gly Leu Ser Pro Asp Leu Pro Ile Cys Lys     610 615 620 Glu Gln Val Gln Ser Cys Gly Pro Pro Pro Glu Leu Leu Asn Gly Asn 625 630 635 640 Val Lys Glu Lys Thr Lys Glu Glu Tyr Gly His Ser Glu Val Val Glu                 645 650 655 Tyr Tyr Cys Asn Pro Arg Phe Leu Met Lys Gly Pro Asn Lys Ile Gln             660 665 670 Cys Val Asp Gly Glu Trp Thr Thr Leu Pro Val Cys Ile Val Glu Glu         675 680 685 Ser Thr Cys Gly Asp Ile Pro Glu Leu Glu His Gly Trp Ala Gln Leu     690 695 700 Ser Ser Pro Pro Tyr Tyr Tyr Gly Asp Ser Val Glu Phe Asn Cys Ser 705 710 715 720 Glu Ser Phe Thr Met Ile Gly His Arg Ser Ile Thr Cys Ile His Gly                 725 730 735 Val Trp Thr Gln Leu Pro Gln Cys Val Ala Ile Asp Lys Leu Lys Lys             740 745 750 Cys Lys Ser Ser Asn Leu Ile Ile Leu Glu Glu His Leu Lys Asn Lys         755 760 765 Lys Glu Phe Asp His Asn Ser Asn Ile Arg Tyr Arg Cys Arg Gly Lys     770 775 780 Glu Gly Trp Ile His Thr Val Cys Ile Asn Gly Arg Trp Asp Pro Glu 785 790 795 800 Val Asn Cys Ser Met Ala Gln Ile Gln Leu Cys Pro Pro Pro Pro Gln                 805 810 815 Ile Pro Asn Ser His Asn Met Thr Thr Thr Leu Asn Tyr Arg Asp Gly             820 825 830 Glu Lys Val Ser Val Leu Cys Gln Glu Asn Tyr Leu Ile Gln Glu Gly         835 840 845 Glu Glu Ile Thr Cys Lys Asp Gly Arg Trp Gln Ser Ile Pro Leu Cys     850 855 860 Val Glu Lys Ile Pro Cys Ser Gln Pro Pro Gln Ile Glu His Gly Thr 865 870 875 880 Ile Asn Ser Ser Arg Ser Ser Gln Glu Ser Tyr Ala His Gly Thr Lys                 885 890 895 Leu Ser Tyr Thr Cys Glu Gly Gly Phe Arg Ile Ser Glu Glu Asn Glu             900 905 910 Thr Cys Tyr Met Gly Lys Trp Ser Ser Pro Pro Gln Cys Glu Gly         915 920 925 Leu Pro Cys Lys Ser Pro Pro Glu Ile Ser His Gly Val Val Ala His     930 935 940 Met Ser Asp Ser Tyr Gln Tyr Gly Glu Glu Val Thr Tyr Lys Cys Phe 945 950 955 960 Glu Gly Phe Gly Ile Asp Gly Pro Ala Ile Ala Lys Cys Leu Gly Glu                 965 970 975 Lys Trp Ser His Pro Pro Ser Cys Ile Lys Thr Asp Cys Leu Ser Leu             980 985 990 Pro Ser Phe Glu Asn Ala Ile Pro Met Gly Glu Lys Lys Asp Val Tyr         995 1000 1005 Lys Ala Gly Glu Gln Val Thr Tyr Thr Cys Ala Thr Tyr Tyr Lys Met    1010 1015 1020 Asp Gly Ala Ser Asn Val Thr Cys Ile Asn Ser Arg Trp Thr Gly Arg 1025 1030 1035 1040 Pro Thr Cys Arg Asp Thr Ser Cys Val Asn Pro Pro Thr Val Gln Asn                1045 1050 1055 Ala Tyr Ile Val Ser Arg Gln Met Ser Lys Tyr Pro Ser Gly Glu Arg            1060 1065 1070 Val Arg Tyr Gln Cys Arg Ser Pro Tyr Glu Met Phe Gly Asp Glu Glu        1075 1080 1085 Val Met Cys Leu Asn Gly Asn Trp Thr Glu Pro Pro Gln Cys Lys Asp    1090 1095 1100 Ser Thr Gly Lys Cys Gly Pro Pro Pro Pro Ile Asp Asn Gly Asp Ile 1105 1110 1115 1120 Thr Ser Phe Pro Leu Ser Val Tyr Ala Pro Ala Ser Ser Val Glu Tyr                1125 1130 1135 Gln Cys Gln Asn Leu Tyr Gln Leu Glu Gly Asn Lys Arg Ile Thr Cys            1140 1145 1150 Arg Asn Gly Gln Trp Ser Glu Pro Pro Lys Cys Leu His Pro Cys Val        1155 1160 1165 Ile Ser Arg Glu Ile Met Glu Asn Tyr Asn Ile Ala Leu Arg Trp Thr    1170 1175 1180 Ala Lys Gln Lys Leu Tyr Ser Arg Thr Gly Glu Ser Val Glu Phe Val 1185 1190 1195 1200 Cys Lys Arg Gly Tyr Arg Leu Ser Ser Arg Ser His Thr Leu Arg Thr                1205 1210 1215 Thr Cys Trp Asp Gly Lys Leu Glu Tyr Pro Thr Cys Ala Lys Arg            1220 1225 1230 <210> 22 <211> 764 <212> PRT <213> Homo sapiens CFB amino acid sequence <400> 22 Met Gly Ser Asn Leu Ser Pro Gln Leu Cys Leu Met Pro Phe Ile Leu   1 5 10 15 Gly Leu Leu Ser Gly Gly Val Thr Thr Thr Pro Trp Ser Leu Ala Arg              20 25 30 Pro Gln Gly Ser Cys Ser Leu Glu Gly Val Glu Ile Lys Gly Gly Ser          35 40 45 Phe Arg Leu Leu Gln Glu Gly Gln Ala Leu Glu Tyr Val Cys Pro Ser      50 55 60 Gly Phe Tyr Pro Tyr Pro Val Gln Thr Arg Thr Cys Arg Ser Thr Gly  65 70 75 80 Ser Trp Ser Thr Leu Lys Thr Gln Asp Gln Lys Thr Val Arg Lys Ala                  85 90 95 Glu Cys Arg Ala Ile His Cys Pro Arg Pro His Asp Phe Glu Asn Gly             100 105 110 Glu Tyr Trp Pro Arg Ser Pro Tyr Tyr Asn Val Ser Asp Glu Ile Ser         115 120 125 Phe His Cys Tyr Asp Gly Tyr Thr Leu Arg Gly Ser Ala Asn Arg Thr     130 135 140 Cys Gln Val Asn Gly Arg Trp Ser Gly Gln Thr Ala Ile Cys Asp Asn 145 150 155 160 Gly Ala Gly Tyr Cys Ser Asn Pro Gly Ile Pro Ile Gly Thr Arg Lys                 165 170 175 Val Gly Ser Gln Tyr Arg Leu Glu Asp Ser Val Thr Tyr His Cys Ser             180 185 190 Arg Gly Leu Thr Leu Arg Gly Ser Gln Arg Arg Thr Cys Gln Glu Gly         195 200 205 Gly Ser Trp Ser Gly Thr Glu Pro Ser Cys Gln Asp Ser Phe Met Tyr     210 215 220 Asp Thr Pro Gln Glu Val Ala Glu Ala Phe Leu Ser Ser Leu Thr Glu 225 230 235 240 Thr Ile Glu Gly Val Asp Ala Glu Asp Gly His Gly Pro Gly Glu Gln                 245 250 255 Gln Lys Arg Lys Ile Val Leu Asp Pro Ser Gly Ser Met Asn Ile Tyr             260 265 270 Leu Val Leu Asp Gly Ser Asp Ser Ile Gly Ala Ser Asn Phe Thr Gly         275 280 285 Ala Lys Lys Cys Leu Val Asn Leu Ile Glu Lys Val Ala Ser Tyr Gly     290 295 300 Val Lys Pro Arg Tyr Gly Leu Val Thr Tyr Ala Thr Tyr Pro Lys Ile 305 310 315 320 Trp Val Lys Val Ser Glu Ala Asp Ser Ser Asn Ala Asp Trp Val Thr                 325 330 335 Lys Gln Leu Asn Glu Ile Asn Tyr Glu Asp His Lys Leu Lys Ser Gly             340 345 350 Thr Asn Thr Lys Lys Ala Leu Gln Ala Val Tyr Ser Met Met Ser Trp         355 360 365 Pro Asp Val Pro Pro Glu Gly Trp Asn Arg Thr Arg His Val Ile     370 375 380 Ile Leu Met Thr Asp Gly Leu His Asn Met Gly Gly Asp Pro Ile Thr 385 390 395 400 Val Ile Asp Glu Ile Arg Asp Leu Leu Tyr Ile Gly Lys Asp Arg Lys                 405 410 415 Asn Pro Arg Glu Asp Tyr Leu Asp Val Tyr Val Phe Gly Val Gly Pro             420 425 430 Leu Val Asn Gln Val Asn Ile Asn Ala Leu Ala Ser Lys Lys Asp Asn         435 440 445 Glu Gln His Val Phe Lys Val Lys Asp Met Glu Asn Leu Glu Asp Val     450 455 460 Phe Tyr Gln Met Ile Asp Glu Ser Gln Ser Leu Ser Leu Cys Gly Met 465 470 475 480 Val Trp Glu His Arg Lys Gly Thr Asp Tyr His Lys Gln Pro Trp Gln                 485 490 495 Ala Lys Ile Ser Val Ile Arg Pro Ser Lys Gly His Glu Ser Cys Met             500 505 510 Gly Ala Val Val Ser Glu Tyr Phe Val Leu Thr Ala Ala His Cys Phe         515 520 525 Thr Val Asp Asp Lys Glu His Ser Ile Lys Val Ser Val Gly Gly Glu     530 535 540 Lys Arg Asp Leu Glu Ile Glu Val Val Leu Phe His Pro Asn Tyr Asn 545 550 555 560 Ile Asn Gly Lys Lys Glu Ala Gly Ile Pro Glu Phe Tyr Asp Tyr Asp                 565 570 575 Val Ala Leu Ile Lys Leu Lys Asn Lys Leu Lys Tyr Gly Gln Thr Ile             580 585 590 Arg Pro Ile Cys Leu Pro Cys Thr Glu Gly Thr Thr Arg Ala Leu Arg         595 600 605 Leu Pro Pro Thr Thr Thr Cys Gln Gln Gln Lys Glu Glu Leu Leu Pro     610 615 620 Ala Gln Asp Ile Lys Ala Leu Phe Val Ser Glu Glu Glu Lys Lys Leu 625 630 635 640 Thr Arg Lys Glu Val Tyr Ile Lys Asn Gly Asp Lys Lys Gly Ser Cys                 645 650 655 Glu Arg Asp Ala Gln Tyr Ala Pro Gly Tyr Asp Lys Val Lys Asp Ile             660 665 670 Ser Glu Val Val Thr Pro Arg Phe Leu Cys Thr Gly Gly Val Ser Pro         675 680 685 Tyr Ala Asp Pro Asn Thr Cys Arg Gly Asp Ser Gly Gly Pro Leu Ile     690 695 700 Val His Lys Arg Ser Ser Phe Ile Gln Val Gly Val Ile Ser Trp Gly 705 710 715 720 Val Val Asp Val Cys Lys Asn Gln Lys Arg Gln Lys Gln Val Pro Ala                 725 730 735 His Ala Arg Asp Phe His Ile Asn Leu Phe Gln Val Leu Pro Trp Leu             740 745 750 Lys Glu Lys Leu Gln Asp Glu Asp Leu Gly Phe Leu         755 760 <210> 23 <211> 1663 <212> PRT <213> Homo sapiens C3 amino acid sequence <400> 23 Met Gly Pro Thr Ser Gly Pro Ser Leu Leu Leu Leu Leu Leu Thr His   1 5 10 15 Leu Pro Leu Ala Leu Gly Ser Pro Met Tyr Ser Ile Ile Thr Pro Asn              20 25 30 Ile Leu Arg Leu Glu Ser Glu Glu Thr Met Val Leu Glu Ala His Asp          35 40 45 Ala Gln Gly Asp Val Pro Thr Val Thr Val His Asp Phe Pro Gly      50 55 60 Lys Lys Leu Val Leu Ser Ser Glu Lys Thr Val Leu Thr Pro Ala Thr  65 70 75 80 Asn His Met Gly Asn Val Thr Phe Thr Ile Pro Ala Asn Arg Glu Phe                  85 90 95 Lys Ser Glu Lys Gly Arg Asn Lys Phe Val Thr Val Gln Ala Thr Phe             100 105 110 Gly Thr Gln Val Val Glu Lys Val Val Leu Val Ser Leu Gln Ser Gly         115 120 125 Tyr Leu Phe Ile Gln Thr Asp Lys Thr Ile Tyr Thr Pro Gly Ser Thr     130 135 140 Val Leu Tyr Arg Ile Phe Thr Val Asn His Lys Leu Leu Pro Val Gly 145 150 155 160 Arg Thr Val Met Val Asn Ile Glu Asn Pro Glu Gly Ile Pro Val Lys                 165 170 175 Gln Asp Ser Leu Ser Ser Gln Asn Gln Leu Gly Val Leu Pro Leu Ser             180 185 190 Trp Asp Ile Pro Glu Leu Val Asn Met Gly Gln Trp Lys Ile Arg Ala         195 200 205 Tyr Tyr Glu Asn Ser Pro Gln Gln Val Phe Ser Thr Glu Phe Glu Val     210 215 220 Lys Glu Tyr Val Leu Pro Ser Phe Glu Val Ile Val Glu Pro Thr Glu 225 230 235 240 Lys Phe Tyr Tyr Ile Tyr Asn Glu Lys Gly Leu Glu Val Thr Ile Thr                 245 250 255 Ala Arg Phe Leu Tyr Gly Lys Lys Val Glu Gly Thr Ala Phe Val Ile             260 265 270 Phe Gly Ile Gln Asp Gly Glu Gln Arg Ile Ser Leu Pro Glu Ser Leu         275 280 285 Lys Arg Ile Pro Ile Glu Asp Gly Ser Gly Glu Val Val Leu Ser Arg     290 295 300 Lys Val Leu Leu Asp Gly Val Gln Asn Pro Arg Ala Glu Asp Leu Val 305 310 315 320 Gly Lys Ser Leu Tyr Val Ser Ala Thr Val Ile Leu His Ser Gly Ser                 325 330 335 Asp Met Val Gln Ala Glu Arg Ser Gly Ile Pro Ile Val Thr Ser Pro             340 345 350 Tyr Gln Ile His Phe Thr Lys Thr Pro Lys Tyr Phe Lys Pro Gly Met         355 360 365 Pro Phe Asp Leu Met Val Phe Val Thr Asn Pro Asp Gly Ser Pro Ala     370 375 380 Tyr Arg Val Val Val Ala Val Gln Gly Glu Asp Thr Val Gln Ser Leu 385 390 395 400 Thr Gln Gly Asp Gly Val Ala Lys Leu Ser Ile Asn Thr His Ser Ser                 405 410 415 Gln Lys Pro Leu Ser Ile Thr Val Arg Thr Lys Lys Gln Glu Leu Ser             420 425 430 Glu Ala Glu Gln Ala Thr Arg Thr Met Gln Ala Leu Pro Tyr Ser Thr         435 440 445 Val Gly Asn Ser Asn Asn Tyr Leu His Leu Ser Val Leu Arg Thr Glu     450 455 460 Leu Arg Pro Gly Glu Thr Leu Asn Val Asn Phe Leu Leu Arg Met Asp 465 470 475 480 Arg Ala His Glu Ala Lys Ile Arg Tyr Tyr Thr Tyr Leu Ile Met Asn                 485 490 495 Lys Gly Arg Leu Leu Lys Ala Gly Arg Gln Val Arg Glu Pro Gly Gln             500 505 510 Asp Leu Val Val Leu Pro Leu Ser Ile Thr Thr Asp Phe Ile Pro Ser         515 520 525 Phe Arg Leu Val Ala Tyr Tyr Thr Leu Ile Gly Ala Ser Gly Gln Arg     530 535 540 Glu Val Val Ala Asp Ser Val Trp Val Asp Val Lys Asp Ser Cys Val 545 550 555 560 Gly Ser Leu Val Val Lys Ser Gly Gln Ser Glu Asp Arg Gln Pro Val                 565 570 575 Pro Gly Gln Gln Met Thr Leu Lys Ile Glu Gly Asp His Gly Ala Arg             580 585 590 Val Val Leu Val Ala Val Asp Lys Val Val Val Leu Asn Lys Lys         595 600 605 Asn Lys Leu Thr Gln Ser Lys Ile Trp Asp Val Val Glu Lys Ala Asp     610 615 620 Ile Gly Cys Thr Pro Gly Ser Gly Lys Asp Tyr Ala Gly Val Phe Ser 625 630 635 640 Asp Ala Gly Leu Thr Phe Thr Ser Ser Gly Gln Gln Thr Ala Gln                 645 650 655 Arg Ala Glu Leu Gln Cys Pro Gln Pro Ala Ala Arg Arg Arg Arg Ser             660 665 670 Val Gln Leu Thr Glu Lys Arg Met Asp Lys Val Gly Lys Tyr Pro Lys         675 680 685 Glu Leu Arg Lys Cys Cys Glu Asp Gly Met Arg Glu Asn Pro Met Arg     690 695 700 Phe Ser Cys Gln Arg Arg Thr Arg Phe Ile Ser Leu Gly Glu Ala Cys 705 710 715 720 Lys Lys Val Phe Leu Asp Cys Cys Asn Tyr Ile Thr Glu Leu Arg Arg                 725 730 735 Gln His Ala Arg Ala Ser His Leu Gly Leu Ala Arg Ser Asn Leu Asp             740 745 750 Glu Asp Ile Ile Ala Glu Glu Asn Ile Val Ser Ser Ser Glu Phe Pro         755 760 765 Glu Ser Trp Leu Trp Asn Val Glu Asp Leu Lys Glu Pro Pro Lys Asn     770 775 780 Gly Ile Ser Thr Lys Leu Met Asn Ile Phe Leu Lys Asp Ser Ile Thr 785 790 795 800 Thr Trp Glu Ile Leu Ala Val Ser Met Ser Asp Lys Lys Gly Ile Cys                 805 810 815 Val Ala Asp Pro Phe Glu Val Thr Val Met Gln Asp Phe Phe Ile Asp             820 825 830 Leu Arg Leu Pro Tyr Ser Val Val Arg Asn Glu Gln Val Glu Ile Arg         835 840 845 Ala Val Leu Tyr Asn Tyr Arg Gln Asn Gln Glu Leu Lys Val Arg Val     850 855 860 Glu Leu Leu His Asn Pro Ala Phe Cys Ser Leu Ala Thr Thr Lys Arg 865 870 875 880 Arg His Gln Gln Thr Val Thr Ile Pro Pro Lys Ser Ser Leu Ser Val                 885 890 895 Pro Tyr Val Ile Val Pro Leu Lys Thr Gly Leu Gln Glu Val Glu Val             900 905 910 Lys Ala Ala Val Tyr His His Phe Ile Ser Asp Gly Val Arg Lys Ser         915 920 925 Leu Lys Val Val Pro Glu Gly Ile Arg Met Asn Lys Thr Val Ala Val     930 935 940 Arg Thr Leu Asp Pro Glu Arg Leu Gly Arg Glu Gly Val Gln Lys Glu 945 950 955 960 Asp Ile Pro Pro Ala Asp Leu Ser Asp Gln Val Pro Asp Thr Glu Ser                 965 970 975 Glu Thr Arg Ile Leu Glu Gln Gly Thr Pro Val Ala Gln Met Thr Glu             980 985 990 Asp Ala Val Asp Ala Glu Arg Leu Lys His Leu Ile Val Thr Pro Ser         995 1000 1005 Gly Cys Gly Glu Gln Asn Met Ile Gly Met Thr Pro Thr Val Ile Ala    1010 1015 1020 Val His Tyr Leu Asp Glu Thr Glu Gln Trp Glu Lys Phe Gly Leu Glu 1025 1030 1035 1040 Lys Arg Gln Gly Ala Leu Glu Leu Ile Lys Lys Gly Tyr Thr Gln Gln                1045 1050 1055 Leu Ala Phe Arg Gln Pro Ser Ser Ala Phe Ala Ala Phe Val Lys Arg            1060 1065 1070 Ala Pro Ser Thr Trp Leu Thr Ala Tyr Val Val Lys Val Phe Ser Leu        1075 1080 1085 Ala Val Asn Leu Ile Ala Ile Asp Ser Gln Val Leu Cys Gly Ala Val    1090 1095 1100 Lys Trp Leu Ile Leu Glu Lys Gln Lys Pro Asp Gly Val Phe Gln Glu 1105 1110 1115 1120 Asp Ala Pro Val Ile His Gln Glu Met Ile Gly Gly Leu Arg Asn Asn                1125 1130 1135 Asn Glu Lys Asp Met Ala Leu Thr Ala Phe Val Leu Ile Ser Leu Gln            1140 1145 1150 Glu Ala Lys Asp Ile Cys Glu Glu Gln Val Asn Ser Leu Pro Gly Ser        1155 1160 1165 Ile Thr Lys Ala Gly Asp Phe Leu Glu Ala Asn Tyr Met Asn Leu Gln    1170 1175 1180 Arg Ser Tyr Thr Val Ala Ile Ala Gly Tyr Ala Leu Ala Gln Met Gly 1185 1190 1195 1200 Arg Leu Lys Gly Pro Leu Leu Asn Lys Phe Leu Thr Thr Ala Lys Asp                1205 1210 1215 Lys Asn Arg Trp Glu Asp Pro Gly Lys Gln Leu Tyr Asn Val Glu Ala            1220 1225 1230 Thr Ser Tyr Ala Leu Leu Ala Leu Leu Gln Leu Lys Asp Phe Asp Phe        1235 1240 1245 Val Pro Pro Val Val Arg Trp Leu Asn Glu Gln Arg Tyr Tyr Gly Gly    1250 1255 1260 Gly Tyr Gly Ser Thr Gln Ala Thr Phe Met Val Phe Gln Ala Leu Ala 1265 1270 1275 1280 Gln Tyr Gln Lys Asp Ala Pro Asp His Gln Glu Leu Asn Leu Asp Val                1285 1290 1295 Ser Leu Gln Leu Pro Ser Arg Ser Ser Lys Ile Thr His Arg Ile His            1300 1305 1310 Trp Glu Ser Ala Ser Leu Leu Arg Ser Glu Glu Thr Lys Glu Asn Glu        1315 1320 1325 Gly Phe Thr Val Thr Ala Glu Gly Lys Gly Gln Gly Thr Leu Ser Val    1330 1335 1340 Val Thr Met Tyr His Ala Lys Ala Lys Asp Gln Leu Thr Cys Asn Lys 1345 1350 1355 1360 Phe Asp Leu Lys Val Thr Ile Lys Pro Ala Pro Glu Thr Glu Lys Arg                1365 1370 1375 Pro Gln Asp Ala Lys Asn Thr Met Ile Leu Glu Ile Cys Thr Arg Tyr            1380 1385 1390 Arg Gly Asp Gln Asp Ala Thr Met Ser Ile Leu Asp Ile Ser Met Met        1395 1400 1405 Thr Gly Phe Ala Pro Asp Thr Asp Asp Leu Lys Gln Leu Ala Asn Gly    1410 1415 1420 Val Asp Arg Tyr Ile Ser Lys Tyr Glu Leu Asp Lys Ala Phe Ser Asp 1425 1430 1435 1440 Arg Asn Thr Leu Ile Ile Tyr Leu Asp Lys Val Ser His Ser Glu Asp                1445 1450 1455 Asp Cys Leu Ala Phe Lys Val His Gln Tyr Phe Asn Val Glu Leu Ile            1460 1465 1470 Gln Pro Gly Ala Val Lys Val Tyr Ala Tyr Tyr Asn Leu Glu Glu Ser        1475 1480 1485 Cys Thr Arg Phe Tyr His Pro Glu Lys Glu Asp Gly Lys Leu Asn Lys    1490 1495 1500 Leu Cys Arg Asp Glu Leu Cys Arg Cys Ala Glu Glu Asn Cys Phe Ile 1505 1510 1515 1520 Gln Lys Ser Asp Asp Lys Val Thr Leu Glu Glu Arg Leu Asp Lys Ala                1525 1530 1535 Cys Glu Pro Gly Val Asp Tyr Val Tyr Lys Thr Arg Leu Val Lys Val            1540 1545 1550 Gln Leu Ser Asn Asp Phe Asp Glu Tyr Ile Met Ala Ile Glu Gln Thr        1555 1560 1565 Ile Lys Ser Gly Ser Asp Glu Val Gln Val Gly Gln Gln Arg Thr Phe    1570 1575 1580 Ile Ser Pro Ile Lys Cys Arg Glu Ala Leu Lys Leu Glu Glu Lys Lys 1585 1590 1595 1600 His Tyr Leu Met Trp Gly Leu Ser Ser Asp Phe Trp Gly Glu Lys Pro                1605 1610 1615 Asn Leu Ser Tyr Ile Ile Gly Lys Asp Thr Trp Val Glu His Trp Pro            1620 1625 1630 Glu Glu Asp Glu Cys Gln Asp Glu Glu Asn Gln Lys Gln Cys Gln Asp        1635 1640 1645 Leu Gly Ala Phe Thr Glu Ser Met Val Val Phe Gly Cys Pro Asn    1650 1655 1660 <210> 24 <211> 583 <212> PRT <213> Homo sapiens CFI amino acid sequence <400> 24 Met Lys Leu Leu His Val Phe Leu Leu Phe Leu Cys Phe His Leu Arg   1 5 10 15 Phe Cys Lys Val Thr Tyr Thr Ser Gln Glu Asp Leu Val Glu Lys Lys              20 25 30 Cys Leu Ala Lys Lys Tyr Thr His Leu Ser Cys Asp Lys Val Phe Cys          35 40 45 Gln Pro Trp Gln Arg Cys Ile Glu Gly Thr Cys Val Cys Lys Leu Pro      50 55 60 Tyr Gln Cys Pro Lys Asn Gly Thr Ala Val Cys Ala Thr Asn Arg Arg  65 70 75 80 Ser Phe Pro Thr Tyr Cys Gln Gln Lys Ser Leu Glu Cys Leu His Pro                  85 90 95 Gly Thr Lys Phe Leu Asn Asn Gly Thr Cys Thr Ala Glu Gly Lys Phe             100 105 110 Ser Val Ser Leu Lys His Gly Asn Thr Asp Ser Glu Gly Ile Val Glu         115 120 125 Val Lys Leu Val Asp Gln Asp Lys Thr Met Phe Ile Cys Lys Ser Ser     130 135 140 Trp Ser Met Arg Glu Ala Asn Val Ala Cys Leu Asp Leu Gly Phe Gln 145 150 155 160 Gln Gly Ala Asp Thr Gln Arg Arg Phe Lys Leu Ser Asp Leu Ser Ile                 165 170 175 Asn Ser Thr Glu Cys Leu His Val His Cys Arg Gly Leu Glu Thr Ser             180 185 190 Leu Ala Glu Cys Thr Phe Thr Lys Arg Arg Thr Met Gly Tyr Gln Asp         195 200 205 Phe Ala Asp Val Val Cys Tyr Thr Gln Lys Ala Asp Ser Pro Met Asp     210 215 220 Asp Phe Phe Gln Cys Val Asn Gly Lys Tyr Ile Ser Gln Met Lys Ala 225 230 235 240 Cys Asp Gly Ile Asn Asp Cys Gly Asp Gln Ser Asp Glu Leu Cys Cys                 245 250 255 Lys Ala Cys Gln Gly Lys Gly Phe His Cys Lys Ser Gly Val Cys Ile             260 265 270 Pro Ser Gln Tyr Gln Cys Asn Gly Glu Val Asp Cys Ile Thr Gly Glu         275 280 285 Asp Glu Val Gly Cys Ala Gly Phe Ala Ser Val Ala Gln Glu Glu Thr     290 295 300 Glu Ile Leu Thr Ala Asp Met Asp Ala Glu Arg Arg Arg Ile Lys Ser 305 310 315 320 Leu Leu Pro Lys Leu Ser Cys Gly Val Lys Asn Arg Met His Ile Arg                 325 330 335 Arg Lys Arg Ile Val Gly Gly Lys Arg Ala Gln Leu Gly Asp Leu Pro             340 345 350 Trp Gln Val Ala Ile Lys Asp Ala Ser Gly Ile Thr Cys Gly Gly Ile         355 360 365 Tyr Ile Gly Gly Cys Trp Ile Leu Thr Ala Ala His Cys Leu Arg Ala     370 375 380 Ser Lys Thr His Arg Tyr Gln Ile Trp Thr Thr Val Val Asp Trp Ile 385 390 395 400 His Pro Asp Leu Lys Arg Ile Val Ile Glu Tyr Val Asp Arg Ile Ile                 405 410 415 Phe His Glu Asn Tyr Asn Ala Gly Thr Tyr Gln Asn Asp Ile Ala Leu             420 425 430 Ile Glu Met Lys Lys Asp Gly Asn Lys Lys Asp Cys Glu Leu Pro Arg         435 440 445 Ser Ile Pro Ala Cys Val Pro Trp Ser Pro Tyr Leu Phe Gln Pro Asn     450 455 460 Asp Thr Cys Ile Val Ser Gly Trp Gly Arg Glu Lys Asp Asn Glu Arg 465 470 475 480 Val Phe Ser Leu Gln Trp Gly Glu Val Lys Leu Ile Ser Asn Cys Ser                 485 490 495 Lys Phe Tyr Gly Asn Arg Phe Tyr Glu Lys Glu Met Glu Cys Ala Gly             500 505 510 Thr Tyr Asp Gly Ser Ile Asp Ala Cys Lys Gly Asp Ser Gly Gly Pro         515 520 525 Leu Val Cys Met Asp Ala Asn Asn Val Thr Tyr Val Trp Gly Val Val     530 535 540 Ser Trp Gly Glu Asn Cys Gly Lys Pro Glu Phe Pro Gly Val Tyr Thr 545 550 555 560 Lys Val Ala Asn Tyr Phe Asp Trp Ile Ser Tyr His Val Gly Arg Pro                 565 570 575 Phe Ile Ser Gln Tyr Asn Val             580 <210> 25 <211> 3696 <212> DNA <213> Homo sapiens CHF nucleotides sequence <400> 25 atgagacttc tagcaaagat tatttgcctt atgttatggg ctatttgtgt agcagaagat 60 tgcaatgaac ttcctccaag aagaaataca gaaattctga caggttcctg gtctgaccaa 120 acatatccag aaggcaccca ggctatctat aaatgccgcc ctggatatag atctcttgga 180 aatgtaataa tggtatgcag gaagggagaa tgggttgctc ttaatccatt aaggaaatgt 240 cagaaaaggc cctgtggaca tcctggagat actccttttg gtacttttac ccttacagga 300 ggaaatgtgt ttgaatatgg tgtaaaagct gtgtatacat gtaatgaggg gtatcaattg 360 ctaggtgaga ttaattaccg tgaatgtgac acagatggat ggaccaatga tattcctata 420 tgtgaagttg tgaagtgttt accagtgaca gcaccagaga atggaaaaat tgtcagtagt 480 gcaatggaac cagatcggga ataccatttt ggacaagcag tacggtttgt atgtaactca 540 ggctacaaga ttgaaggaga tgaagaaatg cattgttcag acgatggttt ttggagtaaa 600 gagaaaccaa agtgtgtgga aatttcatgc aaatccccag atgttataaa tggatctcct 660 atatctcaga agattattta taaggagaat gaacgatttc aatataaatg taacatgggt 720 tatgaataca gtgaaagagg agatgctgta tgcactgaat ctggatggcg tccgttgcct 780 tcatgtgaag aaaaatcatg tgataatcct tatattccaa atggtgacta ctcaccttta 840 aggattaaac acagaactgg agatgaaatc acgtaccagt gtagaaatgg tttttatcct 900 gcaacccggg gaaatacagc aaaatgcaca agtactggct ggatacctgc tccgagatgt 960 accttgaaac cttgtgatta tccagacatt aaacatggag gtctatatca tgagaatatg 1020 cgtagaccat actttccagt agctgtagga aaatattact cctattactg tgatgaacat 1080 tttgagactc cgtcaggaag ttactgggat cacattcatt gcacacaaga tggatggtcg 1140 ccagcagtac catgcctcag aaaatgttat tttccttatt tggaaaatgg atataatcaa 1200 aatcatggaa gaaagtttgt acagggtaaa tctatagacg ttgcctgcca tcctggctac 1260 gctcttccaa aagcgcagac cacagttaca tgtatggaga atggctggtc tcctactccc 1320 agatgcatcc gtgtcaaaac atgttccaaa tcaagtatag atattgagaa tgggtttatt 1380 tctgaatctc agtatacata tgccttaaaa gaaaaagcga aatatcaatg caaactagga 1440 tatgtaacag cagatggtga aacatcagga tcaattacat gtgggaaaga tggatggtca 1500 gctcaaccca cgtgcattaa atcttgtgat atcccagtat ttatgaatgc cagaactaaa 1560 aatgacttca catggtttaa gctgaatgac acattggact atgaatgcca tgatggttat 1620 gaaagcaata ctggaagcac cactggttcc atagtgtgtg gttacaatgg ttggtctgat 1680 ttacccatat gttatgaaag agaatgcgaa cttcctaaaa tagatgtaca cttagttcct 1740 gatcgcaaga aagaccagta taaagttgga gaggtgttga aattctcctg caaaccagga 1800 tttacatag ttggacctaa ttccgttcag tgctaccact ttggattgtc tcctgacctc 1860 ccaatatgta aagagcaagt acaatcatgt ggtccacctc ctgaactcct caatgggaat 1920 gttaaggaaa aaacgaaaga agaatatgga cacagtgaag tggtggaata ttattgcaat 1980 cctagatttc taatgaaggg acctaataaa attcaatgtg ttgatggaga gtggacaact 2040 ttaccagtgt gtattgtgga ggagagtacc tgtggagata tacctgaact tgaacatggc 2100 tgggcccagc tttcttcccc tccttattac tatggagatt cagtggaatt caattgctca 2160 gaatcattta caatgattgg acacagatca attacgtgta ttcatggagt atggacccaa 2220 cttccccagt gtgtggcaat agataaactt aagaagtgca aatcatcaaa tttaattata 2280 cttgaggaac atttaaaaaa caagaaggaa ttcgatcata attctaacat aaggtacaga 2340 tgtagaggaa aagaaggatg gatacacaca gtctgcataa atggaagatg ggatccagaa 2400 gtgaactgct caatggcaca aatacaatta tgcccacctc cacctcagat tcccaattct 2460 cacaatatga caaccacact gaattatcgg gatggagaaa aagtatctgt tctttgccaa 2520 gaaaattatc taattcagga aggagaagaa attacatgca aagatggaag atggcagtca 2580 ataccactct gtgttgaaaa aattccatgt tcacaaccac ctcagataga acacggaacc 2640 attaattcat ccaggtcttc acaagaaagt tatgcacatg ggactaaatt gagttatact 2700 tgtgagggtg gtttcaggat atctgaagaa aatgaaacaa catgctacat gggaaaatgg 2760 agttctccac ctcagtgtga aggccttcct tgtaaatctc cacctgagat ttctcatggt 2820 gttgtagctc acatgtcaga cagttatcag tatggagaag aagttacgta caaatgtttt 2880 gaaggttttg gaattgatgg gcctgcaatt gcaaaatgct taggagaaaa atggtctcac 2940 cctccatcat gcataaaaac agattgtctc agtttaccta gctttgaaaa tgccataccc 3000 atgggagaga agaaggatgt gtataaggcg ggtgagcaag tgacttacac ttgtgcaaca 3060 tattacaaaa tggatggagc cagtaatgta acatgcatta atagcagatg gacaggaagg 3120 ccaacatgca gagacacctc ctgtgtgaat ccgcccacag tacaaaatgc ttatatagtg 3180 tcgagacaga tgagtaaata tccatctggt gagagagtac gttatcaatg taggagccct 3240 tatgaaatgt ttggggatga agaagtgatg tgtttaaatg gaaactggac ggaaccacct 3300 caatgcaaag attctacagg aaaatgtggg ccccctccac ctattgacaa tggggacatt 3360 acttcattcc cgttgtcagt atatgctcca gcttcatcag ttgagtacca atgccagaac 3420 ttgtatcaac ttgagggtaa caagcgaata acatgtagaa atggacaatg gtcagaacca 3480 ccaaaatgct tacatccgtg tgtaatatcc cgagaaatta tggaaaatta taacatagca 3540 ttaaggtgga cagccaaaca gaagctttat tcgagaacag gtgaatcagt tgaatttgtg 3600 tgtaaacggg gatatcgtct ttcatcacgt tctcacacat tgcgaacaac atgttgggat 3660 gggaaactgg agtatccaac ttgtgcaaaa agatag 3696 <210> 26 <211> 2295 <212> DNA <213> Homo sapiens CFB nucleotides sequence <400> 26 atggggagca atctcagccc ccaactctgc ctgatgccct ttatcttggg cctcttgtct 60 ggaggtgtga ccaccactcc atggtctttg gcccggcccc agggatcctg ctctctggag 120 ggggtagaga tcaaaggcgg ctccttccga cttctccaag agggccaggc actggagtac 180 gtgtgtcctt ctggcttcta cccgtaccct gtgcagacac gtacctgcag atctacgggg 240 tcctggagca ccctgaagac tcaagaccaa aagactgtca ggaaggcaga gtgcagagca 300 atccactgtc caagaccaca cgacttcgag aacggggaat actggccccg gtctccctac 360 tacaatgtga gtgatgagat ctctttccac tgctatgacg gttacactct ccggggctct 420 gccaatcgca cctgccaagt gaatggccga tggagtgggc agacagcgat ctgtgacaac 480 ggagcggggt actgctccaa cccgggcatc cccattggca caaggaaggt gggcagccag 540 taccgccttg aagacagcgt cacctaccac tgcagccggg ggcttaccct gcgtggctcc 600 cgcggcgaa cgtgtcagga aggtggctct tggagcggga cggagccttc ctgccaagac 660 tccttcatgt acgacacccc tcaagaggtg gccgaagctt tcctgtcttc cctgacagag 720 accatagaag gagtcgatgc tgaggatggg cacggcccag gggaacaaca gaagcggaag 780 atcgtcctgg acccttcagg ctccatgaac atctacctgg tgctagatgg atcagacagc 840 attggggcca gcaacttcac aggagccaaa aagtgtctag tcaacttaat tgagaaggtg 900 gcaagttatg gtgtgaagcc aagatatggt ctagtgacat atgccacata ccccaaaatt 960 tgggtcaaag tgtctgaagc agacagcagt aatgcagact gggtcacgaa gcagctcaat 1020 gaaatcaatt atgaagacca caagttgaag tcagggacta acaccaagaa ggccctccag 1080 gcagtgtaca gcatgatgag ctggccagat gacgtccctc ctgaaggctg gaaccgcacc 1140 cgccatgtca tcatcctcat gactgatgga ttgcacaaca tgggcgggga cccaattact 1200 gtcattgatg agatccggga cttgctatac attggcaagg atcgcaaaaa cccaagggag 1260 gattatctgg atgtctatgt gtttggggtc gggcctttgg tgaaccaagt gaacatcaat 1320 gctttggctt ccaagaaaga caatgagcaa catgtgttca aagtcaagga tatggaaaac 1380 ctggaagatg ttttctacca aatgatcgat gaaagccagt ctctgagtct ctgtggcatg 1440 gtttgggaac acaggaaggg taccgattac cacaagcaac catggcaggc caagatctca 1500 gtcattcgcc cttcaaaggg acacgagagc tgtatggggg ctgtggtgtc tgagtacttt 1560 gtgctgacag cagcacattg tttcactgtg gatgacaagg aacactcaat caaggtcagc 1620 gtaggagggg agaagcggga cctggagata gaagtagtcc tatttcaccc caactacaac 1680 attaatggga aaaaagaagc aggaattcct gaattttatg actatgacgt tgccctgatc 1740 aagctcaaga ataagctgaa atatggccag actatcaggc ccatttgtct cccctgcacc 1800 gagggaacaa ctcgagcttt gaggcttcct ccaactacca cttgccagca acaaaaggaa 1860 gagctgctcc ctgcacagga tatcaaagct ctgtttgtgt ctgaggagga gaaaaagctg 1920 actcggaagg aggtctacat caagaatggg gataagaaag gcagctgtga gagagatgct 1980 caatatgccc caggctatga caaagtcaag gacatctcag aggtggtcac ccctcggttc 2040 ctttgtactg gaggagtgag tccctatgct gaccccaata cttgcagagg tgattctggc 2100 ggccccttga tagttcacaa gagaagtcgt ttcattcaag ttggtgtaat cagctgggga 2160 gtagtggatg tctgcaaaaa ccagaagcgg caaaagcagg tacctgctca cgcccgagac 2220 tttcacatca acctctttca agtgctgccc tggctgaagg agaaactcca agatgaggat 2280 ttgggttttc tataa 2295 <210> 27 <211> 4992 <212> DNA <213> Homo sapiens C3 nucleotides sequence <400> 27 atgggaccca cctcaggtcc cagcctgctg ctcctgctac taacccacct ccccctggct 60 ctggggagtc ccatgtactc tatcatcacc cccaacatct tgcggctgga gagcgaggag 120 accatggtgc tggaggccca cgacgcgcaa ggggatgttc cagtcactgt tactgtccac 180 gacttcccag gcaaaaaact agtgctgtcc agtgagaaga ctgtgctgac ccctgccacc 240 aaccacatgg gcaacgtcac cttcacgatc ccagccaaca gggagttcaa gtcagaaaag 300 gggcgcaaca agttcgtgac cgtgcaggcc accttcggga cccaagtggt ggagaaggtg 360 gtgctggtca gcctgcagag cgggtacctc ttcatccaga cagacaagac catctacacc 420 cctggctcca cagttctcta tcggatcttc accgtcaacc acaagctgct acccgtgggc 480 cggacggtca tggtcaacat tgagaacccg gaaggcatcc cggtcaagca ggactccttg 540 tcttctcaga accagcttgg cgtcttgccc ttgtcttggg acattccgga actcgtcaac 600 atgggccagt ggaagatccg agcctactat gaaaactcac cacagcaggt cttctccact 660 gagtttgagg tgaaggagta cgtgctgccc agtttcgagg tcatagtgga gcctacagag 720 aaattctact acatctataa cgagaagggc ctggaggtca ccatcaccgc caggttcctc 780 tacgggaaga aagtggaggg aactgccttt gtcatcttcg ggatccagga tggcgaacag 840 aggatttccc tgcctgaatc cctcaagcgc attccgattg aggatggctc gggggaggtt 900 gtgctgagcc ggaaggtact gctggacggg gtgcagaacc cccgagcaga agacctggtg 960 gggaagtctt tgtacgtgtc tgccaccgtc atcttgcact caggcagtga catggtgcag 1020 gcagagcgca gcgggatccc catcgtgacc tctccctacc agatccactt caccaagaca 1080 cccaagtact tcaaaccagg aatgcccttt gacctcatgg tgttcgtgac gaaccctgat 1140 ggctctccag cctaccgagt ccccgtggca gtccagggcg aggacactgt gcagtctcta 1200 acccagggag atggcgtggc caaactcagc atcaacacac accccagcca gaagcccttg 1260 agcatcacgg tgcgcacgaa gaagcaggag ctctcggagg cagagcaggc taccaggacc 1320 atgcaggctc tgccctacag caccgtgggc aactccaaca attacctgca tctctcagtg 1380 ctacgtacag agctcagacc cggggagacc ctcaacgtca acttcctcct gcgaatggac 1440 cgcgcccacg aggccaagat ccgctactac acctacctga tcatgaacaa gggcaggctg 1500 ttgaaggcgg gacgccaggt gcgagagccc ggccaggacc tggtggtgct gcccctgtcc 1560 atcaccaccg acttcatccc ttccttccgc ctggtggcgt actacacgct gatcggtgcc 1620 agcggccaga gggaggtggt ggccgactcc gtgtgggtgg acgtcaagga ctcctgcgtg 1680 ggctcgctgg tggtaaaaag cggccagtca gaagaccggc agcctgtacc tgggcagcag 1740 atgaccctga agatagaggg tgaccacggg gcccgggtgg tactggtggc cgtggacaag 1800 ggcgtgttcg tgctgaataa gaagaacaaa ctgacgcaga gtaagatctg ggacgtggtg 1860 gagaaggcag acatcggctg caccccgggc agtgggaagg attacgccgg tgtcttctcc 1920 gacgcagggc tgaccttcac gagcagcagt ggccagcaga ccgcccagag ggcagaactt 1980 cagtgcccgc agccagccgc ccgccgacgc cgttccgtgc agctcacgga gaagcgaatg 2040 gacaaagtcg gcaagtaccc caaggagctg cgcaagtgct gcgaggacgg catgcgggag 2100 aaccccatga ggttctcgtg ccagcgccgg acccgtttca tctccctggg cgaggcgtgc 2160 aagaaggtct tcctggactg ctgcaactac atcacagagc tgcggcggca gcacgcgcgg 2220 gccagccacc tgggcctggc caggagtaac ctggatgagg acatcattgc agaagagaac 2280 atcgtttccc gaagtgagtt cccagagagc tggctgtgga acgttgagga cttgaaagag 2340 ccaccgaaaa atggaatctc tacgaagctc atgaatatat ttttgaaaga ctccatcacc 2400 acgtgggaga ttctggctgt gagcatgtcg gacaagaaag ggatctgtgt ggcagacccc 2460 ttcgaggtca cagtaatgca ggacttcttc atcgacctgc ggctacccta ctctgttgtt 2520 cgaaacgagc aggtggaaat ccgagccgtt ctctacaatt accggcagaa ccaagagctc 2580 aaggtgaggg tggaactact ccacaatcca gccttctgca gcctggccac caccaagagg 2640 cgtcaccagc agaccgtaac catccccccc aagtcctcgt tgtccgttcc atatgtcatc 2700 gtgccgctaa agaccggcct gcaggaagtg gaagtcaagg ctgctgtcta ccatcatttc 2760 atcagtgacg gtgtcaggaa gtccctgaag gtcgtgccgg aaggaatcag aatgaacaaa 2820 actgtggctg ttcgcaccct ggatccagaa cgcctgggcc gtgaaggagt gcagaaagag 2880 gacatcccac ctgcagacct cagtgaccaa gtcccggaca ccgagtctga gaccagaatt 2940 ctcctgcaag ggaccccagt ggcccagatg acagaggatg ccgtcgacgc ggaacggctg 3000 aagcacctca ttgtgacccc ctcgggctgc ggggaacaga acatgatcgg catgacgccc 3060 acggtcatcg ctgtgcatta cctggatgaa acggagcagt gggagaagtt cggcctagag 3120 aagcggcagg gggccttgga gctcatcaag aaggggtaca cccagcagct ggccttcaga 3180 caacccagct ctgcctttgc ggccttcgtg aaacgggcac ccagcacctg gctgaccgcc 3240 tacgtggtca aggtcttctc tctggctgtc aacctcatcg ccatcgactc ccaagtcctc 3300 tgcggggctg ttaaatggct gatcctggag aagcagaagc ccgacggggt cttccaggag 3360 gggcgcccg tgatacacca agaaatgatt ggtggattac ggaacaacaa cgagaaagac 3420 atggccctca cggcctttgt tctcatctcg ctgcaggagg ctaaagatat ttgcgaggag 3480 caggtcaaca gcctgccagg cagcatcact aaagcaggag acttccttga agccaactac 3540 atgaacctac agagatccta cactgtggcc attgctggct atgctctggc ccagatgggc 3600 aggctgaagg ggcctcttct taacaaattt ctgaccacag ccaaagataa gaaccgctgg 3660 gaggaccctg gtaagcagct ctacaacgtg gaggccacat cctatgccct cttggcccta 3720 ctgcagctaa aagactttga ctttgtgcct cccgtcgtgc gttggctcaa tgaacagaga 3780 tactacggtg gtggctatgg ctctacccag gccaccttca tggtgttcca agccttggct 3840 caataccaaa aggacgcccc tgaccaccag gaactgaacc ttgatgtgtc cctccaactg 3900 cccagccgca gctccaagat cacccaccgt atccactggg aatctgccag cctcctgcga 3960 tcagaagaga ccaaggaaaa tgagggtttc acagtcacag ctgaaggaaa aggccaaggc 4020 accttgtcgg tggtgacaat gtaccatgct aaggccaaag atcaactcac ctgtaataaa 4080 ttcgacctca aggtcaccat aaaaccagca ccggaaacag aaaagaggcc tcaggatgcc 4140 aagaacacta tgatccttga gatctgtacc aggtaccggg gagaccagga tgccactatg 4200 tctatattgg acatatccat gatgactggc tttgctccag acacatatga cctgaagcag 4260 ctggccaatg gtgttgacag atacatctcc aagtatgagc tggacaaagc cttctccgat 4320 aggaacaccc tcatcatcta cctggacaag gtctcacact ctgaggatga ctgtctagct 4380 ttcaaagttc accaatactt taatgtagag cttatccagc ctggagcagt caaggtctac 4440 gcctattaca acctggagga aagctgtacc cggttctacc atccggaaaa ggaggatgga 4500 aagctgaaca agctctgccg tgatgaactg tgccgctgtg ctgaggagaa ttgcttcata 4560 caaaagtcgg atgacaaggt caccctggaa gaacggctgg acaaggcctg tgagccagga 4620 gtggactatg tgtacaagac ccgactggtc aaggttcagc tgtccaatga ctttgacgag 4680 tacatcatgg ccattgagca gaccatcaag tcaggctcgg atgaggtgca ggttggacag 4740 cagcgcacgt tcatcagccc catcaagtgc agagaagccc tgaagctgga ggagaagaaa 4800 cactacctca tgtggggtct ctcctccgat ttctggggag agaagcccaa cctcagctac 4860 atcatcggga aggacacttg ggtggagcac tggcccgagg aggacgaatg ccaagacgaa 4920 gagaccaga aacaatgcca ggacctcggc gccttcaccg agagcatggt tgtctttggg 4980 tgccccaact ga 4992 <210> 28 <211> 1752 <212> DNA <213> Homo sapiens CFI nucleotides sequence <400> 28 atgaagcttc ttcatgtttt cctgttattt ctgtgcttcc acttaaggtt ttgcaaggtc 60 acttatacat ctcaagagga tctggtggag aaaaagtgct tagcaaaaaa atatactcac 120 ctctcctgcg ataaagtctt ctgccagcca tggcagagat gcattgaggg cacctgtgtt 180 tgtaaactac cgtatcagtg cccaaagaat ggcactgcag tgtgtgcaac taacaggaga 240 agcttcccaa catactgtca acaaaagagt ttggaatgtc ttcatccagg gacaaagttt 300 ttaaataacg gaacatgcac agccgaagga aagtttagtg tttccttgaa gcatggaaat 360 acagattcag agggaatagt tgaagtaaaa cttgtggacc aagataagac aatgttcata 420 tgcaaaagca gctggagcat gagggaagcc aacgtggcct gccttgacct tgggtttcaa 480 caaggtgctg atactcaaag aaggtttaag ttgtctgatc tctctataaa ttccactgaa 540 tgtctacatg tgcattgccg aggattagag accagtttgg ctgaatgtac ttttactaag 600 agaagaacta tgggttacca ggatttcgct gatgtggttt gttatacaca gaaagcagat 660 tctccaatgg atgacttctt tcagtgtgtg aatgggaaat acatttctca gatgaaagcc 720 tgtgatggta tcaatgattg tggagaccaa agtgatgaac tgtgttgtaa agcatgccaa 780 ggcaaaggct tccattgcaa atcgggtgtt tgcattccaa gccagtatca atgcaatggt 840 gaggtggact gcattacagg ggaagatgaa gttggctgtg caggctttgc atctgtggct 900 caagaagaaa cagaaatttt gactgctgac atggatgcag aaagaagacg gataaaatca 960 ttattaccta aactatcttg tggagttaaa aacagaatgc acattcgaag gaaacgaatt 1020 gtgggaggaa agcgagcaca actgggagac ctcccatggc aggtggcaat taaggatgcc 1080 agtggaatca cctgtggggg aatttatatt ggtggctgtt ggattctgac tgctgcacat 1140 tgtctcagag ccagtaaaac tcatcgttac caaatatgga caacagtagt agactggata 1200 caccccgacc ttaaacgtat agtaattgaa tacgtggata gaattatttt ccatgaaaac 1260 tacaatgcag gcacttacca aaatgacatc gctttgattg aaatgaaaaa agacggaaac 1320 aaaaaagatt gtgagctgcc tcgttccatc cctgcctgtg tcccctggtc tccttaccta 1380 ttccaaccta atgatacatg catcgtttct ggctggggac gagaaaaaga taacgaaaga 1440 gtcttttcac ttcagtgggg tgaagttaaa ctaataagca actgctctaa gttttacgga 1500 aatcgtttct atgaaaaaga aatggaatgt gcaggtacat atgatggttc catcgatgcc 1560 tgtaaagggg actctggagg ccccttagtc tgtatggatg ccaacaatgt gacttatgtc 1620 tggggtgttg tgagttgggg ggaaaactgt ggaaaaccag agttcccagg tgtttacacc 1680 aaagtggcca attattttga ctggattagc taccatgtag gaaggccttt tatttctcag 1740 tacaatgtat aa 1752

Claims (14)

Claims 1. A composition for the detection of macular degeneration comprising an agent capable of detecting a mutation consisting of one or a combination of two or more selected from the group of mutations of the following proteins:
The CFH proteins Y56C, L77S, D798E, F176L, G1110A;
The C3 protein R1134Q, A636S, C3 H16Q;
G208S, V716L of CFB protein; And
M204R of CFI protein. The method according to claim 1,
Wherein the agent is a primer pair or a probe that specifically binds to a gene encoding the protein. 3. The method of claim 2,
Wherein said primer pair is selected from the group consisting of SEQ ID NOs: 1 and 2, SEQ ID NOs: 3 and 4, SEQ ID NOs: 5 and 6, SEQ ID NOs: 7 and 8, SEQ ID NOs: 9 and 10, SEQ ID NOs: 11 and 12, SEQ ID NOs: , SEQ ID NOS: 17 and 18, SEQ ID NOS: 19 and 20, respectively. The method according to claim 1,
Wherein the agent is an antibody specific to the protein. A kit for the diagnosis of macular degeneration comprising the composition of any one of claims 1 to 4. 6. The method of claim 5,
Wherein the kit is an RT-PCR kit, a microarray chip kit, or a protein chip kit. 6. The method of claim 5,
Wherein the RT-PCR kit comprises a primer pair that specifically binds to a gene encoding the protein. 8. The method of claim 7,
Wherein said primer pair is selected from the group consisting of SEQ ID NOs: 1 and 2, SEQ ID NOs: 3 and 4, SEQ ID NOs: 5 and 6, SEQ ID NOs: 7 and 8, SEQ ID NOs: 9 and 10, SEQ ID NOs: 11 and 12, SEQ ID NOs: , SEQ ID NOS: 17 and 18, SEQ ID NOS: 19 and 20, respectively. 6. The method of claim 5,
Wherein the microarray chip kit comprises a probe specifically binding to a gene encoding the protein. 6. The method of claim 5,
Wherein the protein chip kit comprises an antibody specific for the protein. (i) performing PCR using the primer pairs of SEQ ID NOS: 1 and 2;
(ii) pyrosequencing using the sequencing primers of SEQ ID NOS: 3 and 4 for the PCR products obtained in step (i); and detecting the mutation of the gene associated with the risk of macular degeneration . 12. The method of claim 11,
Wherein the gene associated with the risk of macular degeneration is a complement factor H (CFH). 13. The method of claim 12,
Wherein the amino acid mutation by the CFH gene mutation is CFH Y56C or L77S. A mutation in which amino acid 56 of SEQ ID NO: 21 is substituted with Tyr to Cys; A mutation in which amino acid 77 is substituted with Leu to Ser; A mutation in which amino acid 798 is substituted with Asp to Glu; A mutation in which amino acid 176 is substituted from Phe to Leu; A mutation in which amino acid 1110 is replaced by Ala in Gly;
A mutation in which amino acid 208 of SEQ ID NO: 22 is substituted with Gly to Ser; A mutation in which amino acid 716 is substituted by Val to Leu;
A mutation in which amino acid 1134 of SEQ ID NO: 23 is substituted from Arg to Gln; A mutant in which amino acid 636 is substituted with Ala to Ser, a mutant in which amino acid 16 is substituted from His to Gln; And
A mutation in which the amino acid at position 204 of SEQ ID NO: 24 is replaced by Met at Arg; and a mutant wherein at least one mutation is selected from the group consisting of:

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