CN100354302C - SIP gene and its coded protein and uses - Google Patents

Abstract

The present invention relates to SIP gene, the SIP protein encoded by the gene, and the application of the gene and protein in medicine. The protein can inhibit tumor growth and proliferation, and can inhibit the expression of luciferase initiated by the cyclinD1 promoter.

Description

SIP gene and encoded protein thereof and application

Technical field

The present invention relates to the SIP gene, the sip protein of this genes encoding, and described gene and the application of albumen in medical science.Described albumen can suppress growth of tumor and propagation, the cyclinD1 promotor is started luciferase expression play restraining effect.

Background technology

Some albumen can suppress the growth and the propagation of tumour cell, finds that the albumen of new inhibition tumor growth and propagation and their gene of coding can be tumor treatment new ways and means is provided.

Summary of the invention

The present invention has found a kind of growth of the cancer cells that suppresses mammary cancer etc. and the albumen of propagation, cloned this proteic gene of coding, made up the host cell that contains this expression carrier and contain this expression vector, and find that this albumen can suppress the growth and the propagation of tumour cells such as mammary cancer, based on above discovery, finished the present invention.

Therefore, in one aspect, the invention provides the growth that can suppress tumour cell and the albumen of propagation, especially recombinant protein, this albumen has the aminoacid sequence shown in the sequence 2 in the sequence table, perhaps by the one or more amino acid whose disappearance in the described aminoacid sequence, replace or increase and the aminoacid sequence that obtains.

In yet another aspect, the invention provides the above-mentioned proteic gene of coding, it be have the base sequence shown in the sequence 1 DNA or with the DNA of described DNA hybridize under stringent condition.

In yet another aspect, the invention provides and contain said gene or encode above-mentioned proteic expression carrier.

In yet another aspect, the invention provides by above-mentioned expression vector transformed host cells.

In yet another aspect, the invention provides the method for producing recombinant protein of the present invention, this method comprises the following steps:

Cultivate above-mentioned transformed host cells, make transformant produce recombinant protein of the present invention; With

From culture, isolate recombinant protein.

In yet another aspect, the invention provides at above-mentioned proteic antibody, the specific antibody that it is to use described recombinant protein to produce as immunogen.It can be mono-clonal or polyclonal antibody.

In yet another aspect, the invention provides and detect described proteic test kit, it contains above-mentioned specific antibody, can carry out antigen-antibody reaction, is used to detect have the albumen shown in the sequence 2.

In addition, the invention provides the probe diagnostics test kit that is used to detect the nucleotide sequence shown in the sequence 1, it contain with nucleotide sequence 1 in partial nucleotide sequence complementary nucleotide sequence.It can be a dna fragmentation with 15-300 base.By the method for probe hybridization, be used to detect the expression of the mRNA that contains described nucleotide sequence, this mRNA is translated into the albumen that contains the aminoacid sequence shown in the sequence 2.

In addition, it is right to the invention provides a primer, be used for carrying out pcr amplification, primer is as follows: Primer1:5 '-tgc tct aga gcc acc atg gac tac aaggac gac gat gac aag gaa ttc atg gcc cag gtc ctg cac-3 '

Primer2：5’-cgc?gga?tcc?cta?ctc?ttc?tgc?cga?gga-3’。

The present invention also provides the application in the medicine of preparation treatment tumour of said gene, albumen or antibody.

The accompanying drawing summary

Fig. 1 is that Northem Blot result shows: gene transcripts conforms to its total length size.And in 8 kinds of tissues of the mankind, expression is arranged all.Conform to the information biology prediction.

Fig. 2 is the polyacrylamide gel electrophoresis figure (SDS-PAGE) of GST-SIP fusion rotein expression and purification in e. coli bl21.90kDa is proteinic molecular weight.The arrow indication is isolated protein band.

Fig. 3 is the expression figure (Western blot) of sip protein in eukaryotic cell MCF-7.One anti-is mouse anti human Flag monoclonal antibody, and the two anti-anti-mouse horseradish peroxidase of rabbit (HRP) link coupled of preserving for this laboratory two resist.1 negative contrast, 2 is cell pyrolysis liquid.PcDNA3.1 (-)-SIP transfection of flag label of MCF-7 cell.Protein level is with anti-flag antibody test.The cell lysates of negative control and MCF-7 is represented in the 1-2 road respectively.

Fig. 4 is the Subcellular Localization of sip protein.Green is GFP, the karyon that blueness is dyed for DAPI.The Subcellular Localization of SIP is in tenuigenin.

Fig. 5 detects the influence of the SIP of different concns to breast cancer cell propagation for mtt assay.X-coordinate is the transfection time, and ordinate zou is the 570nm absorbance.In every prescription post, be v-25ng from left to right successively, SIP-25ng, v-50ng, SIP-50ng, v-100ng, SIP-100ng.

Fig. 6 is SIP starts luciferase expression to the cyclinD1 promotor effect.X-coordinate is the different deletant of cyclinD1 promotor.Ordinate zou is a Firefly/Renilla ratio.Fig. 7 is the structural representation of SIP, and two grey square frames are wherein served as reasons Coils2 ＜http://smart.embl-heidelberg.de/help/smartglossary. shtml 〉The coiled coil of program determination zone and 6 black box be by SEG ＜http://smart.embl-heidelberg.de/help/smartglossary.shtml 〉The low segment of measuring of composition complicacy.

SIP (SRC-1 interacting protein) is the SRC-1 by one of p160 family member The new gene that method by yeast two-hybrid screens. We are from people's mammary gland cDNA library In obtain the ORF of this gene by the method for PCR. Be cloned into pcDNA3.1 (-) eukaryotic expression In the carrier. For ease of detecting Flag label on its N termination. SIP is again KIAA1518, FLJ20004, DKFZp434N161. Be positioned chromosome 19p13.2. Total length 5358bp. Expressing protein～90kD. ORF:2580bp. 859 amino acid of encoding.

It is carried out bioinformatic analysis, find its wide spectrum expression. Comprise Ankyrin repeat Domain.

(1) SIP expressing information:

CDNA source: normal prostatic; Testis; Colon; Colon tumor, RER+; Pooled-Skin; Neuroblastoma COT 10 NORMALIZED; Primary pulmonary cystic fibrosis epithelial cell; Fat Fat; Malignant mela noma, lymphatic metastasis; White matter; Cochlea; Bone marrow matrix; Glioblastoma (pooled); Five kinds merge sarcoma, comprise MLS, single-shot fibroma, malignant fibrous Histocytoma, the stromal tumors tumour of stomach and intestine, and celiothelioma; Small cell carcinoma; Placenta; Lymph; Three kinds merge meningioma; Full brain; Moderate differentiation adenocarcinoma of endometrium, 3 kinds merge tumour; Normally Colon; Chondrosarcoma lung transfer cell line; Medulloblastoma; Tumour; Ovary; The B cell, slow The property lymphocytic leukemia; Epithelioma; Melanoma; Breast; The slurries papillary carcinoma, senior, 2 kinds Merge tumour; Normal testis; Whole embryo mainly is body; Ovarian carcinoma; Prostate; Sympathetic trunk; Liver and spleen; Meningioma; Normal prostatic; Kidney neoplasms; Iris; Crystalline lens; Olfactory epithelium; Placenta; Neck; Normal placenta; Normal endometrium, middle secretory phase, the 23rd cycle Day; Melanocyte; Merge human melanocytes, fetal rhythm, and gravid uterus; Kidney; Gland cancer; Having seal guards against The poorly differentiated gland cancer of cell characteristic; Merge; Normal coloring look epithelium; Melanoma, high MDR is (thin Born of the same parents system); Epithelial cancer cell line; Neuroblastoma; Cervical cancer tumer line; Hippocampus; The brain that merges, Lung, testis; Optic nerve; Cervix tumor; Fetal pancreas (4 Pooled Donors, 18-20 Week, stratagene); The fetus eyes; Old and feeble fibroblast; Ascites; Lung; On the elementary lung Chrotoplast; Tuberculosis kitchen range fibrillatable; The metastatic chondrosarcoma; Chondrosarcoma; Bad differone Endometrium Gland cancer, 2 kinds merge tumour; The uterus; Squamous cell carcinoma; The multiple sclerosis pathology; Insulinoma; Spleen; Squamous cell carcinoma, clone; Retinoblastoma; Mix (pool of 40 RNAs); Interior life Chondroma clone; Subchondral bone; Heart; Lung; Benign tumour; Duodenal adenocarcinoma, clone; Endometrium, gland cell system; Blood; People's pulmonary epithelial cells; The eyes leading portion; The dorsal root nerve Joint; Teratocarcinoma, clone; The RPE/ choroid; Clone; The purifying Langerhans' islands; Smooth muscle Knurl; Germinal center B cell; MDA; Pancreas; Synovial membrane; Chest muscle (mastectomy with Afterwards); Thyroid gland; The neurinoma tumour; Tumor of prostate.

(2) on the amino acid sequence basis, by ExPASy and interPro its major function is tied Functional domain a: Ankyrin is found in the prediction of structure territory. Detail See Figure and table: Fig. 1 is the structural representation of SIP, and two grey square frames are wherein served as reasonsCoils2 <http://smart.embl-heidelberg.de/help/smart glossary.shtml>The coiled coil of program determination zone and 6 black box be bySEG <http://smart.embl-heidelberg.de/help/smartglossary.shtml>The group of measuring Become the low segment of complexity.

Method	AccNumber	ahortName	Iocation-1	Evalue	Iocation-2	Evalue	Iocation-3	Evalue	Iocation-4	Evalue
											FPrintScan	PR01415	ANKYRIN	T[675-687]	2.8	T[687-699]	2.8
HMMPfam	PF00023	Ank	T[674-707]	0.00026	T[746-778]	7.00E-11	T[779-812]	3.90E-06	T[813-833]	0.008
											HMMSmart	SM00248	ANK	T[674-704]	0.0045	T[708-741]	8.00E+02	T[746-775]	6.30E-05	T[779-809]	0.024
ProfileScan	PS50088	ANK_REPEAT	T[674-699]	9.725	T[748-778]	12743
											ProfileScan	PS50297	ANK_REP_REGION	T[668-836]	31.086

The polypeptide of equivalence on the albumen of the application of the invention or the immunology can obtain it and resist Body, antibody is used for detection and the purifying of described albumen. Can utilize albumen of the present invention or its one Divide amino acid sequence originally to produce antibody as immunity. Can be moving by antibody being inoculated to the host Thing and the conventional method that reclaims serum produce polyclonal antibody. Can also pass through conventional hybridization knurl side Method produces monoclonal antibody.

Utilize mtt assay to detect the SIP of variable concentrations to the impact of MCF-7 tumor cell proliferation, Found that GIBC has the effect of inhibition tumor cell propagation.

Therefore, albumen of the present invention can be used for treating tumour, especially breast cancer.

Embodiment

Embodiment 1: with the gene of PCR method from human breast carcinoma cDNA library clone coding sip protein

With mammary cancer cDNA library is template, and Flag label on SIP gene N termination carries out pcr amplification with following primer:

Primer1：5’-tgc?tct?aga?gcc?acc?atg?gac?tac?aag?gacgac?gat?gac?aag?gaa?ttc?atg?gcc?cag?gtcctg?cac-3’

Primer2：5’-cgc?gga?tcc?cta?ctc?ttc?tgc?cga?gga-3’

The condition of amplified reaction: in the reaction volume of 50 μ L, contain 50mmol/L KCl, 10mmol/L Tris-Cl, (PH:8.5), 1.5mmol/L MgCl ₂, 200 μ mol/L dNTP, 10pmol primer, the pfu archaeal dna polymerase of 0.25U (Takara company product).Press 30 cycles of following conditioned response: 94 ℃ of 30sec; 59 ℃ of 30sec; 72 ℃ of 2min 30sec.Amplified production is with the test kit purifying of QIAGEN company, behind XbaI and BamHI double digestion, is cloned in pcDNA3.1 (-) carrier for expression of eukaryon that same enzyme cuts.The dna sequence analysis result shows that the dna sequence dna of PCR product conforms to expection.

Embodiment 2:Northern Blot

The mark of probe and crossover operation are with reference to the Spotlight of Clontech company ^TMRandomPrimer Labeling Kit test kit specification sheets.Process is as follows:

(1) probe mark: 25ng-1 μ gDNA is dissolved in the 20 μ L volumes, 90-100 ℃ 2-3 minute, rapidly ice bath is 5 minutes, add 5 μ L[α- ³²P] the RandomPrimers and Reaction Buffer Mix of dCTP mark, moisturizing to 50 μ L is hatched 5-10min for 37 ℃.

(2) hybridization: 68 ℃ of preheating hybridization solutions, the nylon membrane that is adsorbed with nucleic acid is put into the hybridization bottle that contains hybridization solution, add the hybridization solution 0.1mL/cm of the salmon sperm DNA that contains 100 μ g/mL ², 68 ℃ of prehybridization 30min.The 20ng/mL probe is sneaked into hybridization solution, 95 ℃ 2-5 minute, ice bath 5 minutes.Add the hybridization bottle, hybridized 1 hour for 68 ℃.

(3) wash film: washed film 30-40 minute with Wash Buffer 1 room temperature, washed film 40 minutes with 250 ℃ of Wash Buffer.

(4) colour developing: take out film with tweezers, wash and do Wash Buffer 2, cover with preservative film ,-70 ℃ of exposures were developed photographic fixing after 1-3 days.

Expression and the purifying of embodiment 3:SIP in prokaryotic cell prokaryocyte

PGEX-4T-3-SIP can be in e. coli strain bl21 the fusion rotein of 30 ℃ of abduction delivering GST-SIP, at first prepare the competent cell of BL21: the picking mono-clonal is in an amount of substratum, and 37 ℃ of 250rpm are cultured to A ₆₀₀0.4-0.5, centrifugal 15 minutes of 2500g, standby.1-50ng pGEX-4T-3-SIP is transformed in the competent cell of above-mentioned preparation, carries out purifying with Glutathione Sepharose 4B, the step of purifying is as follows:

(1) getting mono-clonal cultivated 4-5 hour in 2-3mL 2YTA substratum.Be transferred to grow overnight in the Erlenmeyer flask of 100mL.

(2) be transferred in the Erlenmeyer flask of 500mL and cultivated 3-5 hour.

(3) add 30 ℃ of 0.5mL 1M IPTG, cultivated 3-6 hour.

(4) 10 minutes centrifugal collecting cells of 3000rpm.

(5) use the 25mLPBS re-suspended cell.Ultrasonic 10 minutes.

(6) add 1.25mL 20%Triton X-100, mixing 1 hour.

(7) 1000rpm 10 minutes is centrifugal, adds 0.5mL 50%slurry of Glutathione

Sepharose 4B, room temperature 30 minutes.

(8) 1000rpm 5 minutes is centrifugal, and PBS gives a baby a bath on the third day after its birth time.

(9) with 0.5mL wash-out Buffer wash-out, supernatant is collected in of short duration centrifugal back.

The expression of embodiment 4:SIP in eukaryotic cell

Western Blot grows up on the basis of protein electrophorese and solid-phase immunoassay.Its ultimate principle is antigen antibody reaction.The cell protein extract is transferred to (nitrocellulose filter is adopted in this experiment) on the solid support after polyacrylamide gel electrophoresis separates.(the anti-people Flag of this laboratory mice monoclonal antibody Sigma), is hatched with horseradish peroxidase (HRP) link coupled two anti-(it is anti-that the anti-mouse of rabbit two is preserved in this laboratory) again, and colour developing can obtain the western blot figure of differential protein molecule with special one anti-reaction.This method has detect specific antigenic advantage from mixes antigen, so this experiment is used to detect the expression behind the SIP gene transfection MCF-7 cell that has the Flag label.

1) transient transfection experiment:

In transfection the day before yesterday with the MCF-7 cell inoculation in the 10cm culture plate, inoculum density is 2 * 10 ⁶Individual cells/well, every hole adds nutrient solution 10ml.After 24 hours, cell grows to when covering the about 50-60% of culture hole floorage, can prepare transfection.The following solution of preparation in the EP pipe: solution A: 24 μ g pcDNA3.1 (-)-SIP expression plasmid are dissolved in the unparalleled anti-DMEM nutrient solution of 1.5mL serum-free.Solution B: 48 μ L liposome solutions (lipofectamine 2000 reagent) are joined in the unparalleled anti-DMEM nutrient solution of 1.5mL serum-free.Room temperature left standstill 5 minutes.Then solution A, B are mixed, room temperature left standstill 20 minutes.With serum-free with do not contain two anti-DMEM nutrient solution washed cells.With adding the unparalleled anti-DMEM nutrient solution of 12mL serum-free in the mixed solution of above-mentioned solution A, B, add to cell surface behind the mixing.37 ℃, hatch 4-6 hour under the 5%CO2 condition after, replace aforementioned nutrient solution with the DMEM nutrient solution that normally contains 10% foetal calf serum again, continue culturing cell to 48 hour.

2) extraction of total protein of cell:

Cultured cells makes it to disperse through 0.25% trysinization, and 4 ℃ of low-speed centrifugals remove supernatant, and precipitation is placed 30min on ice, and the PBS with precooling washes per 10 2 times again ⁶Cell adds lysate 100 μ L, makes the abundant cracking 30min of cell, and again in 4 ℃, the centrifugal 15min of 20000 * g collects supernatant.

3) the Bradford method is measured the proteic concentration of cell pyrolysis liquid:

At first prepare coomassie brilliant blue staining liquid; Standard bovine serum albumin (BSA) is mixed with the protein standard liquid of 1mg/ml, 10 μ g, 20 μ g, 30 μ g, 40 μ g, 50 μ g, 60 μ g protein standard liquid and 5 μ L testing protein quality samples are added in the 3mL Xylene Brilliant Cyanine G dye liquor respectively, incubated at room 10min measures absorption value at 595nm wavelength place.BSA standard model bioassay standard curve, OD within the specific limits ₅₉₅Be directly proportional with protein concentration.The concentration of sample protein matter is calculated through the typical curve fit equation.4) electrophoresis and immune response: flow process is as follows: A, glue (5% concentrates glue, 10% separation gel); B, 50 μ g protein samples add sample-loading buffer, boil last sample; C, electrophoresis concentrate glue 80V voltage, separation gel 120V voltage; D, commentaries on classics film, 100V, 1.5h; E, sealing, 1h; F, an anti-reaction, are spent the night by 4 ℃; G, TBST wash film, and three times, each 10min; H, two anti-reactions, 1.5h; I, TBST wash film, and three times, each 10min; J, colour developing, exposure.

Embodiment 5:SIP is in intracellular location

The pEGFP-C1 C-terminal albumen fusion vector that utilizes this laboratory to preserve has made up the pEGFP-C1-SIP carrier, and it can express the fusion rotein of GFP-SIP in eukaryotic cell, with this carrier Lipofectin ^TMThe method transfection advance in the MCF-7 cell, cells transfected is carried out creep plate, after 24 hours, gives a baby a bath on the third day after its birth time with PBS, fixes with 4% Paraformaldehyde 96/PBS, room temperature 15 minutes is given a baby a bath on the third day after its birth time with PBS then.With saturatingization of 0.1%Triton X-100/PBS 15 minutes, give a baby a bath on the third day after its birth time with PBS then, add the DAPI of 200 μ L 2.5mg/mL, room temperature effect 30 minutes, give a baby a bath on the third day after its birth time with PBS then, under fluorescent microscope, observe with conventional wavelength of fluorescence and DAPI wavelength respectively, and take pictures.The results are shown in Figure 4, be positioned tenuigenin.

Embodiment 6:MTT method detects the influence of the SIP of different concns to breast cancer cell propagation

MTT (tetramethyl-azo azoles orchid) can be by cellular uptake, and is reduced into a kind of water-fast bluish voilet product first a ceremonial jade-ladle, used in libation by mitochondrial dehydrogenase in the viable cell, and is deposited in the cell, and dead cell does not have this function.The first a ceremonial jade-ladle, used in libation dissolves in dimethyl sulfoxide (DMSO) (DMSO), and solution has maximum absorption at the 570nm place.So the big more representative survivaling cell of this wave band place absorption value amount is many more.Mtt assay is widely used in cytotoxicity experiment, cell growth measurement etc.This experiment utilizes mtt assay to observe the influence of SIP to the MCF-7 growth.Concrete operation method is as follows:

In 96 orifice plates, every hole inoculating cell number is 1 * 10 with the MCF-7 cell inoculation of cultivating logarithmic phase ⁴Individual.Add SIP (25,50,100ng) according to concentration gradient next day, every mass action 6 porocytes, and 37 ℃, 5%CO2 cultivates.After cultivating 1d, 2d, 3d, 4d respectively, the MTT solution that in each cell cultures hole, adds 50 μ lL 1mg/mL, same culture condition effect down took out after 4 hours, liquid in the careful every hole of sucking-off, every hole adds dimethyl sulfoxide (DMSO) 150 μ L, slight vibration 10 minutes fully is dissolved in the dimethyl sulfoxide (DMSO) bluish voilet crystallization.Measure in 96 orifice plates every hole at the light absorption value at 570nm place with automatic microplate reader.Referring to Fig. 5, the MTT experiment shows that SIP plays restraining effect to the MCF-7 growth and proliferation of cell, and relevant with dosage.

Embodiment 7: transient transfection and luciferase (Luciferase) experiment:

Use luciferase as reporter gene, for the controlling element of analyzing gene with measure the method that the genetic transcription activity is provided convenience.This experiment is building up to cyclinD1 gene promoter and luciferase reading frame in the carrier for expression of eukaryon exactly jointly, with this fusion expression plasmid transfectional cell, by measuring the activity of luciferase, thereby the research transcription factor, regulate the effect of albumen to the cyclinD1 gene promoter.It is reporter gene that Photinus pyralis LUC (Firefly) is adopted in experiment, and extra large wild pansy (Renillar) luciferase is an internal reference.With two fluorescence report gene alaysis system test kits that Promega company produces, in same test tube, measure the activity of two kinds of enzymes.And the applied chemistry light-emitting appearance detects.

2) transient transfection experiment:

In transfection the day before yesterday with the MCF-7 cell inoculation in 24 well culture plates, inoculum density is 1-2 * 10 ⁵Individual cells/well, every hole adds nutrient solution 0.5mL.After 24 hours, cell grows to when covering the about 50-60% of culture hole floorage, can prepare transfection.The following solution of preparation in the EP pipe: solution A: 0.8 μ gDNA (cyclinD1 reporter gene plasmid: 200ng; Confidential reference items Renillar plasmid: 10ng; SIP expression plasmid: 600ng or pcDNA3.1 (-) empty carrier plasmid: 600ng), be dissolved in the unparalleled anti-DMEM nutrient solution of 50 μ L serum-frees.Solution B: 2 μ L liposome solutions (lipofectamine 2000 reagent) are joined in the unparalleled anti-DMEM nutrient solution of 50 μ L serum-frees.Room temperature left standstill 5 minutes.Then solution A, B are mixed, room temperature left standstill 20 minutes.With serum-free with do not contain two anti-DMEM nutrient solution washed cells.With adding the unparalleled anti-DMEM nutrient solution of 400 μ L serum-frees in the mixed solution of above-mentioned solution A, B, add to cell surface behind the mixing.Each experimental group repeats 3 holes.37 ℃, 5%CO ₂After hatching 4-6 hour under the condition, replace aforementioned nutrient solution with the DMEM nutrient solution that normally contains 10% foetal calf serum again, continue culturing cell to 48 hour.Lysing cell, the activity of measurement luciferase.

2) with pair activity of the two luciferases of fluorescence report kit genes measurement:

(1) lysing cell:

Clean cell with cold PBS, 100 μ L add each cell cultures hole with 1 * cell pyrolysis liquid (PLB), and incubated at room 15 minutes makes the abundant cracking of cell.Resulting cell pyrolysis liquid can be directly used in fluorescence measurement.

(2) preparation of luciferase substrate:

Photinus pyralis LUC substrate freezing dry powder is dissolved in the 10mL luciferase analysis buffer, and gained solution is LARII, packing ,-70 ℃ of preservations.

Use Stop﹠amp; The Glo damping fluid is diluted to 1 with 50 * extra large wild pansy luciferase substrate *, gained solution is Stop﹠amp; Glo, matching while using.

(3) measuring process:

20 μ L cell pyrolysis liquids are put into 96 hole fluorescence measurement plates, add 100 μ LLARII, measure Photinus pyralis LUC, measuring condition is: postpone 2 seconds, measured 10 seconds.

Add 100 μ LStop﹠amp again; Glo measures extra large wild pansy luciferase, and measuring condition is the same.

Referring to Fig. 6, SIP starts luciferase expression to the cyclinD1 promotor and plays restraining effect.

SIP preface ~ 1

SEQUENCE?LISTING

＜110〉Peking University

＜120〉SIP gene and encoded protein thereof and application

<130>

<160>2

<170>PatentIn?version?3.1

<210>1

<211>2580

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(1)..?(2577)

<223>

<400>1

atg?gcc?cag?gtc?ctg?cac?gtg?cct?gct?ccc?ttc?cca?ggg?acc?cct?ggc 48

Met?Ala?Gln?Val?Leu?His?Val?Pro?Ala?Pro?Phe?Pro?Gly?Thr?Pro?Gly

1 5 10 15

cca?gcc?tcc?cca?cct?gcc?ttc?cct?gcc?aag?gac?ccc?gat?cca?ccc?tac 96

Pro?Ala?Ser?Pro?Pro?Ala?Phe?Pro?Ala?Lys?Asp?Pro?Asp?Pro?Pro?Tyr

20 25 30

tcc?gtg?gag?acc?ccc?tat?ggc?tac?cgc?ctg?gac?ctg?gac?ttc?ctc?aag 144

Ser?Val?Glu?Thr?Pro?Tyr?Gly?Tyr?Arg?Leu?Asp?Leu?Asp?Phe?Leu?Lys

35 40 45

tac?gtg?gat?gac?atc?gag?aag?ggc?cac?acg?ctg?cga?cgc?gtg?gca?gtg 192

Tyr?Val?Asp?Asp?Ile?Glu?Lys?Gly?His?Thr?Leu?Arg?Arg?Val?Ala?Val

50 55 60

cag?cgc?cgc?ccc?cgc?ctg?agc?tcg?ctg?ccc?cgt?ggc?cct?ggc?tcc?tgg 240

Gln?Arg?Arg?Pro?Arg?Leu?Ser?Ser?Leu?Pro?Arg?Gly?Pro?Gly?Ser?Trp

65 70 75 80

tgg?acg?tcc?act?gag?tcg?ctg?tgc?tcc?aat?gcc?agt?ggg?gac?agc?cgc 288

Trp?Thr?Ser?Thr?Glu?Ser?Leu?Cys?Ser?Asn?Ala?Ser?Gly?Asp?Ser?Arg

85 90 95

cac?tca?gcc?tat?tcc?tac?tgc?ggc?cgt?ggc?ttc?tac?cct?cag?tat?ggt 336

His?Ser?Ala?Tyr?Ser?Tyr?Cys?Gly?Arg?Gly?Phe?Tyr?Pro?Gln?Tyr?Gly

100 105 110

gct?ctg?gag?acc?cgc?ggt?ggc?ttc?aat?ccg?cgg?gtg?gag?cgc?acg?ctg 384

Ala?Leu?Glu?Thr?Arg?Gly?Gly?Phe?Asn?Pro?Arg?Val?Glu?Arg?Thr?Leu

115 120 125

ctg?gat?gcc?cgt?cgc?cgt?ctc?gag?gac?cag?gcg?gcc?aca?ccc?acc?ggc 432

Leu?Asp?Ala?Arg?Arg?Arg?Leu?Glu?Asp?Gln?Ala?Ala?Thr?Pro?Thr?Gly

130 135 140

ctg?ggc?tcc?ctg?acc?ccc?agt?gcg?gcc?ggc?tcg?aca?gcc?tcc?ctg?gtg 480

Leu?Gly?Ser?Leu?Thr?Pro?Ser?Ala?Ala?Gly?Ser?Thr?Ala?Ser?Leu?Val

145 150 155 160

ggc?gtg?ggg?ttg?cca?ccc?ccg?aca?cca?cgg?agt?tca?gga?ctg?tcc?aca 528

Gly?Val?Gly?Leu?Pro?Pro?Pro?Thr?Pro?Arg?Ser?Ser?Gly?Leu?Ser?Thr

165 170 175

ccg?gtg?cct?ccc?agt?gcc?ggg?cac?ctg?gcc?cac?gtg?cgg?gag?cag?atg 576

Pro?Val?Pro?Pro?Ser?Ala?Gly?His?Leu?Ala?His?Val?Arg?Glu?Gln?Met

180 185 190

gcg?ggt?gcc?ctg?cgg?aag?ctg?cgg?cag?ctg?gag?gag?cag?gtg?aag?ctg 624

Ala?Gly?Ala?Leu?Arg?Lys?Leu?Arg?Gln?Leu?Glu?Glu?Gln?Val?Lys?Leu

195 200 205

atc?cct?gtg?ctc?cag?gtg?aag?ctc?tcg?gtg?ctc?cag?gag?gaa?aag?cgg 672

Ile?Pro?Val?Leu?Gln?Val?Lys?Leu?Ser?Val?Leu?Gln?Glu?Glu?Lys?Arg

210 215 220

cag?ctc?aca?gta?caa?ctt?aag?agc?cag?aag?ttc?ctg?ggc?cac?ccc?aca 720

Gln?Leu?Thr?Val?Gln?Leu?Lys?Ser?Gln?Lys?Phe?Leu?Gly?His?Pro?Thr

225 230 235 240

gcg?ggc?cgg?ggt?cgc?agc?gag?ctc?tgc?ctg?gac?ctc?ccc?gat?ccc?cca 768

Ala?Gly?Arg?Gly?Arg?Ser?Glu?Leu?Cys?Leu?Asp?Leu?Pro?Asp?Pro?Pro

245 250 255

gag?gac?cca?gtg?gca?ctg?gag?acc?cgg?agt?gtg?ggc?acc?tgg?gtc?cga 816

Glu?Asp?Pro?Val?Ala?Leu?Glu?Thr?Arg?Ser?Val?Gly?Thr?Trp?Val?Arg

260 265 270

gaa?cgg?gac?ttg?ggc?atg?cct?gat?ggg?gag?gct?gcc?ctc?gcc?gcc?aag 864

Glu?Arg?Asp?Leu?Gly?Met?Pro?Asp?Gly?Glu?Ala?Ala?Leu?Ala?Ala?Lys

275 280 285

gtc?gct?gtg?ctg?gag?acc?cag?ctc?aag?aag?gcg?ctt?cag?gag?ctg?cag 912

Val?Ala?Val?Leu?Glu?Thr?Gln?Leu?Lys?Lys?Ala?Leu?Gln?Glu?Leu?Gln

290 295 300

gca?gct?cag?gcc?cgg?cag?gct?gac?ccc?cag?ccc?cag?gcc?tgg?cca?ccg 960

Ala?Ala?Gln?Ala?Arg?Gln?Ala?Asp?Pro?Gln?Pro?Gln?Ala?Trp?Pro?Pro

305 310 315 320

ccg?gac?agc?ccg?gtc?cgc?gtg?gat?aca?gtc?cgg?gtg?gta?gaa?ggg?cca 1008

Pro?Asp?Ser?Pro?Val?Arg?Val?Asp?Thr?Val?Arg?Val?Val?Glu?Gly?Pro

325 330 335

cgg?gag?gtg?gag?gtg?gtg?gcc?agc?aca?gcc?gct?ggc?gcc?ccc?gca?cag 1056

Arg?Glu?Val?Glu?Val?Val?Ala?Ser?Thr?Ala?Ala?Gly?Ala?Pro?Ala?Gln

340 345 350

cgg?gcc?cag?agc?ctg?gag?cct?tac?ggc?aca?ggg?ctg?agg?gcc?ctg?gca 1104

Arg?Ala?Gln?Ser?Leu?Glu?Pro?Tyr?Gly?Thr?Gly?Leu?Arg?Ala?Leu?Ala

355 360 365

atg?cct?ggt?agg?cct?gag?agc?cca?cct?gtg?ttc?cgc?agc?cag?gag?gtg 1152

Met?Pro?Gly?Arg?Pro?Glu?Ser?Pro?Pro?Val?Phe?Arg?Ser?Gln?Glu?Val

370 375 380

gtg?gag?aca?atg?tgc?cca?gtg?ccc?gct?gca?gct?acc?agc?aac?gtc?cat 1200

Val?Glu?Thr?Met?Cys?Pro?Val?Pro?Ala?Ala?Ala?Thr?Ser?Asn?Val?His

385 390 395 400

atg?gtg?aag?aag?att?agc?atc?aca?gag?cga?agc?tgc?gat?gga?gca?gca 1248

Met?Val?Lys?Lys?Ile?Ser?Ile?Thr?Glu?Arg?Ser?Cys?Asp?Gly?Ala?Ala

405 410 415

ggc?ctc?cca?gaa?gtt?cct?gcc?gaa?tcg?tct?tcg?tca?ccc?ecg?ggg?tcc 1296

Gly?Leu?Pro?Glu?Val?Pro?Ala?Glu?Ser?Ser?Ser?Ser?Pro?Pro?Gly?Ser

420 425 430

gag?gta?gcc?tcc?ctt?aca?cag?cct?gag?aag?agc?aca?ggc?cga?gtg?ccc 1344

Glu?Val?Ala?Ser?Leu?Thr?Gln?Pro?Glu?Lys?Ser?Thr?Gly?Arg?Val?Pro

435 440 445

acc?cag?gag?ccc?acc?cac?agg?gag?ccc?acc?agg?caa?gca?gcc?tcc?caa 1392

Thr?Gln?Glu?Pro?Thr?His?Arg?Glu?Pro?Thr?Arg?Gln?Ala?Ala?Ser?Gln

450 455 460

gag?tcc?gag?gag?gcc?ggg?ggc?acc?gca?cct?ccg?ctg?tcc?tcc?cct?cca 1440

Glu?Ser?Glu?Glu?Ala?Gly?Gly?Thr?Ala?Pro?Pro?Leu?Ser?Ser?Pro?Pro

465 470 475 480

ggc?ggg?ccc?ccg?gca?ggc?gtg?cga?tct?atc?atg?aaa?cgg?aaa?gag?gag 1488

Gly?Gly?Pro?Pro?Ala?Gly?Val?Arg?Ser?Ile?Met?Lys?Arg?Lys?Glu?Glu

485 490 495

gtt?gca?gac?ccc?acg?gcc?cac?cgg?agg?agc?ctc?cag?ttc?gtg?ggg?gtc 1536

Val?Ala?Asp?Pro?Thr?Ala?His?Arg?Arg?Ser?Leu?Gln?Phe?Val?Gly?Val

500 505 510

aac?ggc?ggg?tat?gag?tcg?tca?tcc?gag?gac?tcc?agc?aca?gca?gag?aac 1584

Asn?Gly?Gly?Tyr?Glu?Ser?Ser?Ser?Glu?Asp?Ser?Ser?Thr?Ala?Glu?Asn

515 520 525

atc?tca?gac?aac?gac?agc?aca?gag?aac?gag?gcc?cca?gag?ccg?agg?gag 1632

Ile?Ser?Asp?Asn?Asp?Ser?Thr?Glu?Asn?Glu?Ala?Pro?Glu?Pro?Arg?Glu

530 535 540

agg?gtt?ccg?agt?gtg?gcc?gaa?gcc?ccc?cag?ctc?agg?cct?gca?ggg?acg 1680

Arg?Val?Pro?Ser?Val?Ala?Glu?Ala?Pro?Gln?Leu?Arg?Pro?Ala?Gly?Thr

545 550 555 560

gca?gcg?gcc?aag?acc?agc?cgg?cag?gag?tgt?cag?ctg?tct?cga?gaa?tct 1728

Ala?Ala?Ala?Lys?Thr?Ser?Arg?Gln?Glu?Cys?Gln?Leu?Ser?Arg?Glu?Ser

565 570 575

cag?cac?ata?ccc?act?gct?gag?ggg?gca?tca?gga?tca?aac?acg?gag?gag 1776

Gln?His?Ile?Pro?Thr?Ala?Glu?Gly?Ala?Ser?Gly?Ser?Asn?Thr?Glu?Glu

580 585 590

gag?atc?agg?atg?gag?cta?agc?cct?gac?ctc?atc?tca?gcc?tgc?ttg?gcc 1824

Glu?Ile?Arg?Met?Glu?Leu?Ser?Pro?Asp?Leu?Ile?Ser?Ala?Cys?Leu?Ala

595 600 605

ctg?gaa?aag?tac?ctg?gac?aat?ccc?aac?gcc?ctc?aca?gag?cgg?gag?ctg 1872

Leu?Glu?Lys?Tyr?Leu?Asp?Asn?Pro?Asn?Ala?Leu?Thr?Glu?Arg?Glu?Leu

610 615 620

aaa?gtg?gcc?tac?acc?aca?gtg?ctg?cag?gag?tgg?ctg?cgc?ctg?gcc?tgc 1920

Lys?Val?Ala?Tyr?Thr?Thr?Val?Leu?Gln?Glu?Trp?Leu?Arg?Leu?Ala?Cys

625 630 635 640

cgc?agc?gac?gca?cac?ccc?gag?ctg?gtg?cgg?cgg?cac?ctg?gtc?acg?ttc 1968

Arg?Ser?Asp?Ala?His?Pro?Glu?Leu?Val?Arg?Arg?His?Leu?Val?Thr?Phe

645 650 655

cgg?gcc?atg?tct?gcg?cgg?ctg?ctg?gac?tac?gtg?gtc?aac?atc?gcc?gac 2016

Arg?Ala?Met?Ser?Ala?Arg?Leu?Leu?Asp?Tyr?Val?Val?Asn?Ile?Ala?Asp

660 665 670

agc?aac?ggc?aac?aca?gcc?ctg?cac?tac?tcc?gtg?tct?cat?gcc?aac?ttc 2064

Ser?Asn?Gly?Asn?Thr?Ala?Leu?His?Tyr?Ser?Val?Ser?His?Ala?Asn?Phe

675 680 685

ccc?gtg?gtg?cag?cag?ctg?ctc?gac?agc?ggt?gtc?tgc?aag?gtg?gac?aaa 2112

Pro?Val?Val?Gln?Gln?Leu?Leu?Asp?Ser?Gly?Val?Cys?Lys?Val?Asp?Lys

690 695 700

cag?aac?cgt?gct?ggc?tac?agc?cct?att?atg?ctc?acc?gcc?ctg?gcc?acc 2160

Gln?Asn?Arg?Ala?Gly?Tyr?Ser?Pro?Ile?Met?Leu?Thr?Ala?Leu?Ala?Thr

705 710 715 720

ctg?aag?acc?cag?gac?gac?atc?gag?act?gtc?ctt?cag?ctc?ttc?cgg?ctt 2208

Leu?Lys?Thr?Gln?Asp?Asp?Ile?Glu?Thr?Val?Leu?Gln?Leu?Phe?Arg?Leu

725 730 735

ggc?aac?atc?aat?gcc?aaa?gcc?agc?cag?gca?gga?cag?acg?gcc?ctg?atg 2256

Gly?Asn?Ile?Asn?Ala?Lys?Ala?Ser?Gln?Ala?Gly?Gln?Thr?Ala?Leu?Met

740 745 750

ctg?gcc?gtc?agc?cac?ggg?cgg?gtg?gac?gtt?gtc?aaa?gcc?ctg?ctg?gcc 2304

Leu?Ala?Val?Ser?His?Gly?Arg?Val?Asp?Val?Val?Lys?Ala?Leu?Leu?Ala

755 760 765

tgt?gag?gca?gat?gtc?aac?gtg?caa?gat?gat?gac?ggc?tcc?acg?gcc?ctc 2352

Cys?Glu?Ala?Asp?Val?Asn?Val?Gln?Asp?Asp?Asp?Gly?Ser?Thr?Ala?Leu

770 775 780

atg?tgc?gcc?tgt?gag?cac?ggc?cac?aag?gag?atc?gcg?ggg?ctg?ctg?ctg 2400

Met?Cys?Ala?Cys?Glu?His?Gly?His?Lys?Glu?Ile?Ala?Gly?Leu?Leu?Leu

785 790 795 800

gcc?gtg?ccc?agc?tgt?gac?atc?tca?ctc?aca?gat?cgc?gat?ggg?agc?aca 2448

Ala?Val?Pro?Ser?Cys?Asp?Ile?Ser?Leu?Thr?Asp?Arg?Asp?Gly?Ser?Thr

805 810 815

gct?ctg?atg?gtg?gcc?ttg?gac?gca?ggg?cag?agt?gag?att?gcg?tcc?atg 2496

Ala?Leu?Met?Val?Ala?Leu?Asp?Ala?Gly?Gln?Ser?Glu?Ile?Ala?Ser?Met

820 825 830

ctg?tat?tcc?cgc?atg?aac?atc?aag?tgc?tcg?ttt?gcc?cca?atg?tca?gat 2544

Leu?Tyr?Ser?Arg?Met?Asn?Ile?Lys?Cys?Ser?Phe?Ala?Pro?Met?Ser?Asp

835 840 845

gac?gag?agc?cct?aca?tca?tcc?tcg?gca?gaa?gag?tag 2580

Asp?Glu?Ser?Pro?Thr?Ser?Ser?Ser?Ala?Glu?Glu

850 855

<210>2

<211>859

<212>PRT

＜213〉people

<400>2

Met?Ala?Gln?Val?Leu?His?Val?Pro?Ala?Pro?Phe?Pro?Gly?Thr?Pro?Gly

1 5 10 15

Pro?Ala?Ser?Pro?Pro?Ala?Phe?Pro?Ala?Lys?Asp?Pro?Asp?Pro?Pro?Tyr

20 25 30

Ser?Val?Glu?Thr?Pro?Tyr?Gly?Tyr?Arg?Leu?Asp?Leu?Asp?Phe?Leu?Lys

35 40 45

Tyr?Val?Asp?Asp?Ile?Glu?Lys?Gly?His?Thr?Leu?Arg?Arg?Val?Ala?Val

50 55 60

Gln?Arg?Arg?Pro?Arg?Leu?Ser?Ser?Leu?Pro?Arg?Gly?Pro?Gly?Ser?Trp

65 70 75 80

Trp?Thr?Ser?Thr?Glu?Ser?Leu?Cys?Ser?Asn?Ala?Ser?Gly?Asp?Ser?Arg

85 90 95

His?Ser?Ala?Tyr?Ser?Tyr?Cys?Gly?Arg?Gly?Phe?Tyr?Pro?Gln?Tyr?Gly

100 105 110

Ala?Leu?Glu?Thr?Arg?Gly?Gly?Phe?Asn?Pro?Arg?Val?Glu?Arg?Thr?Leu

115 120 125

Leu?Asp?Ala?Arg?Arg?Arg?Leu?Glu?Asp?Gln?Ala?Ala?Thr?Pro?Thr?Gly

130 135 140

Leu?Gly?Ser?Leu?Thr?Pro?Ser?Ala?Ala?Gly?Ser?Thr?Ala?Ser?Leu?Val

145 150 155 160

Gly?Val?Gly?Leu?Pro?Pro?Pro?Thr?Pro?Arg?Ser?Ser?Gly?Leu?Ser?Thr

165 170 175

Pro?Val?Pro?Pro?Ser?Ala?Gly?His?Leu?Ala?His?Val?Arg?Glu?Gln?Met

180 185 190

Ala?Gly?Ala?Leu?Arg?Lys?Leu?Arg?Gln?Leu?Glu?Glu?Gln?Val?Lys?Leu

195 200 205

Ile?Pro?Val?Leu?Gln?Val?Lys?Leu?Ser?Val?Leu?Gln?Glu?Glu?Lys?Arg

210 215 220

Gln?Leu?Thr?Val?Gln?Leu?Lys?Ser?Gln?Lys?Phe?Leu?Gly?His?Pro?Thr

225 230 235 240

Ala?Gly?Arg?Gly?Arg?Ser?Glu?Leu?Cys?Leu?Asp?Leu?Pro?Asp?Pro?Pro

245 250 255

Glu?Asp?Pro?Val?Ala?Leu?Glu?Thr?Arg?Ser?Val?Gly?Thr?Trp?Val?Arg

260 265 270

Glu?Arg?Asp?Leu?Gly?Met?Pro?Asp?Gly?Glu?Ala?Ala?Leu?Ala?Ala?Lys

275 280 285

Val?Ala?Val?Leu?Glu?Thr?Gln?Leu?Lys?Lys?Ala?Leu?Gln?Glu?Leu?Gln

290 295 300

Ala?Ala?Gln?Ala?Arg?Gln?Ala?Asp?Pro?Gln?Pro?Gln?Ala?Trp?Pro?Pro

305 310 315 320

Pro?Asp?Ser?Pro?Val?Arg?Val?Asp?Thr?Val?Arg?Val?Val?Glu?Gly?Pro

325 330 335

Arg?Glu?Val?Glu?Val?Val?Ala?Ser?Thr?Ala?Ala?Gly?Ala?Pro?Ala?Gln

340 345 350

Arg?Ala?Gln?Ser?Leu?Glu?Pro?Tyr?Gly?Thr?Gly?Leu?Arg?Ala?Leu?Ala

355 360 365

Met?Pro?Gly?Arg?Pro?Glu?Ser?Pro?Pro?Val?Phe?Arg?Ser?Gln?Glu?Val

370 375 380

Val?Glu?Thr?Met?Cys?Pro?Val?Pro?Ala?Ala?Ala?Thr?Ser?Asn?Val?His

385 390 395 400

Met?Val?Lys?Lys?Ile?Ser?Ile?Thr?Glu?Arg?Ser?Cys?Asp?Gly?Ala?Ala

405 410 415

Gly?Leu?Pro?Glu?Val?Pro?Ala?Glu?Ser?Ser?Ser?Ser?Pro?Pro?Gly?Ser

420 425 430

Glu?Val?Ala?Ser?Leu?Thr?Gln?Pro?Glu?Lys?Ser?Thr?Gly?Arg?Val?Pro

435 440 445

Thr?Gln?Glu?Pro?Thr?His?Arg?Glu?Pro?Thr?Arg?Gln?Ala?Ala?Ser?Gln

450 455 460

Glu?Ser?Glu?Glu?Ala?Gly?Gly?Thr?Ala?Pro?Pro?Leu?Ser?Ser?Pro?Pro

465 470 475 480

Gly?Gly?Pro?Pro?Ala?Gly?Val?Arg?Ser?Ile?Met?Lys?Arg?Lys?Glu?Glu

485 490 495

Val?Ala?Asp?Pro?Thr?Ala?His?Arg?Arg?Ser?Leu?Gln?Phe?Val?Gly?Val

500 505 510

Asn?Gly?Gly?Tyr?Glu?Ser?Ser?Ser?Glu?Asp?Ser?Ser?Thr?Ala?Glu?Asn

515 520 525

Ile?Ser?Asp?Asn?Asp?Ser?Thr?Glu?Asn?Glu?Ala?Pro?Glu?Pro?Arg?Glu

530 535 540

Arg?Val?Pro?Ser?Val?Ala?Glu?Ala?Pro?Gln?Leu?Arg?Pro?Ala?Gly?Thr

545 550 555 560

Ala?Ala?Ala?Lys?Thr?Ser?Arg?Gln?Glu?Cys?Gln?Leu?Ser?Arg?Glu?Ser

565 570 575

Gln?His?Ile?Pro?Thr?Ala?Glu?Gly?Ala?Ser?Gly?Ser?Asn?Thr?Glu?Glu

580 585 590

Glu?Ile?Arg?Met?Glu?Leu?Ser?Pro?Asp?Leu?Ile?Ser?Ala?Cys?Leu?Ala

595 600 605

Leu?Glu?Lys?Tyr?Leu?Asp?Asn?Pro?Asn?Ala?Leu?Thr?Glu?Arg?Glu?Leu

610 615 620

Lys?Val?Ala?Tyr?Thr?Thr?Val?Leu?Gln?Glu?Trp?Leu?Arg?Leu?Ala?Cys

625 630 635 640

Arg?Ser?Asp?Ala?His?Pro?Glu?Leu?Val?Arg?Arg?His?Leu?Val?Thr?Phe

645 650 655

Arg?Ala?Met?Ser?Ala?Arg?Leu?Leu?Asp?Tyr?Val?Val?Asn?Ile?Ala?Asp

660 665 670

Ser?Asn?Gly?Asn?Thr?Ala?Leu?His?Tyr?Ser?Val?Ser?His?Ala?Asn?Phe

675 680 685

Pro?Val?Val?Gln?Gln?Leu?Leu?Asp?Ser?Gly?Val?Cys?Lys?Val?Asp?Lys

690 695 700

Gln?Asn?Arg?Ala?Gly?Tyr?Ser?Pro?Ile?Met?Leu?Thr?Ala?Leu?Ala?Thr

705 710 715 720

Leu?Lys?Thr?Gln?Asp?Asp?Ile?Glu?Thr?Val?Leu?Gln?Leu?Phe?Arg?Leu

725 730 735

Gly?Asn?Ile?Asn?Ala?Lys?Ala?Ser?Gln?Ala?Gly?Gln?Thr?Ala?Leu?Met

740 745 750

Leu?Ala?Val?Ser?His?Gly?Arg?Val?Asp?Val?Val?Lys?Ala?Leu?Leu?Ala

755 760 765

Cys?Glu?Ala?Asp?Val?Asn?Val?Gln?Asp?Asp?Asp?Gly?Ser?Thr?Ala?Leu

770 775 780

Met?Cys?Ala?Cys?Glu?His?Gly?His?Lys?Glu?Ile?Ala?Gly?Leu?Leu?Leu

785 790 795 800

Ala?Val?Pro?Ser?Cys?Asp?Ile?Ser?Leu?Thr?Asp?Arg?Asp?Gly?Ser?Thr

805 810 815

Ala?Leu?Met?Val?Ala?Leu?Asp?Ala?Gly?Gln?Ser?Glu?Ile?Ala?Ser?Met

820 825 830

Leu?Tyr?Ser?Arg?Met?Asn?Ile?Lys?Cys?Ser?Phe?Ala?Pro?Met?Ser?Asp

835 840 845

Asp?Glu?Ser?Pro?Thr?Ser?Ser?Ser?Ala?Glu?Glu

850 855

Claims (2)

1. Use of a protein with an amino acid sequence as shown in Sequence 2 in the Sequence Listing in the preparation of a medicament for treating breast cancer. 2. Use of the gene encoding the protein of claim 1 whose DNA sequence is shown in Sequence 1 in the Sequence Listing in the preparation of a drug for treating breast cancer.

Images