CN100436595C - Method for diagnosis and treatment of hair eipidermal tumor using human's CYLD gene and its coded products - Google Patents

Method for diagnosis and treatment of hair eipidermal tumor using human's CYLD gene and its coded products Download PDF

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CN100436595C
CN100436595C CNB021108579A CN02110857A CN100436595C CN 100436595 C CN100436595 C CN 100436595C CN B021108579 A CNB021108579 A CN B021108579A CN 02110857 A CN02110857 A CN 02110857A CN 100436595 C CN100436595 C CN 100436595C
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ser
glu
gly
val
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CN1438322A (en
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孔祥银
郑广勇
黄伟
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Shanghai Institute of Nutrition and Health of CAS
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Shanghai Institutes for Biological Sciences SIBS of CAS
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Abstract

The present invention discloses a method for diagnosing trichoepithelioma, which comprises the following procedures that whether aberrance exists or not is checked with the cylindromatosis tumor suppressor protein (CYLD) genes, the transcripts and/or the protein of individuals compared with normal crowds; the existence of the aberrance indicates that the possibility that the individuals have trichoepithelioma is bigger than the normal crowds. The present invention also discloses a corresponding test reagent box, a method for treating trichoepithelioma and a medicine composition.

Description

Human's CYLD gene and coded product thereof the purposes in the preparation diagnostic reagent
Technical field
The present invention relates to biotechnology and medical field.More specifically, the present invention relates to utilize the method for human's CYLD gene and coded product diagnosis and treatment trichoepithelioma, and contain CYLD gene and/or proteic pharmaceutical composition.
Background technology
Trichoepithelioma is a kind of dermatoma.It is fallen ill at preadolescence usually, good sending out, especially nasolabial groove and upper lip in face, and the back, scalp, neck, trunk, four limbs also can be got involved.Its skin decreases and is semisphere or taper shape, and quality is solid, and is translucent, and yellow or incarnadine, bigger sometimes skin decrease and show visible telangiectasis.Trichoepithelioma not only influences beauty treatment, because it makes the back, and scalp, neck, trunk, four limbs are got involved, so severe patient also can be met difficulty in life.This disease is an autosome dominant disease, and the mechanism of its morbidity is not clear at present.Still there is not at present satisfied methods of treatment for this disease.
Yet, up to now, still not fully do not disclose the definite reason of trichoepithelioma, also nobody discloses trichoepithelioma and there is direct dependency in certain albumen.In addition, this area also lacks the effective ways of early diagnosis trichoepithelioma disease and the effective means of non-operative treatment trichoepithelioma.
CYLD is a kind of known protein, and its essential information is as follows:
English: Gene from:Homo sapiens chromosome 16working draft sequencesegment.
NCBI：Contig NT_030834
GENE：Gene from：Homo sapiens chromosome 16 working draft sequencesegment.gi|17488444：243550-308161
The dna sequence dna of CYLD is shown in SEQ ID NO:1, and ORF is positioned at the 392-3262 position, 956 amino acid whose protein of the total length of encoding (SEQ ID NO:2).Other information of CYLD can be from Http: //www.ncbi.nlm.nih.gov obtains.
Once the be in the news sudden change of its exon of CYLD gene has caused generation cylindromatous, but does not still have the relation of any article report CYLD gene and trichoepithelioma in the world.The hereditary mechanistic information of trichoepithelioma can obtain from the OMIM of NCBI.
Therefore, this area presses for the effective ways of new diagnosis of exploitation and treatment trichoepithelioma, and relevant medicine, diagnostic techniques and reagent.
Summary of the invention
One object of the present invention just provides a kind of method and detection kit of new diagnosis (especially early diagnosis) trichoepithelioma.
Another object of the present invention provides a kind of method of new treatment trichoepithelioma.
A further object of the present invention provides a kind of pharmaceutical composition for the treatment of trichoepithelioma.
In a first aspect of the present invention, a kind of method that the trichoepithelioma susceptibility of individuality is diagnosed is provided, it comprises step:
Detect this individual CYLD gene, transcript and/or albumen, and compare with normal CYLD gene, transcript and/or albumen,
The possibility that there are differences with regard to showing this individuality trouble trichoepithelioma is higher than normal population.
Preferably, detected is gene or the transcript of CYLD, and with normal CYLD nucleotide sequence comparing difference.More preferably, described difference is selected from: the 1853rd A disappearance among the SEQ ID NO:1.
In a second aspect of the present invention, a kind of method for the treatment of trichoepithelioma is provided, it comprises step: the normal CYLD albumen of using safe and effective amount to the patient of the described treatment of needs.Preferably, CYLD albumen is locally applied to affected area.
In a third aspect of the present invention, provide a kind of CYLD albumen aspect pharmaceutical compositions purposes and corresponding pharmaceutical compositions, it contains the people CYLD albumen and the pharmaceutically acceptable carrier of safe and effective amount.Preferably, it is an ointment.
In a fourth aspect of the present invention, a kind of test kit that detects trichoepithelioma is provided, it comprises the primer of specific amplification CYLD gene or transcript.Preferably, it also contain with mutable site bonded probe and/or identification the mutational site enzyme.
Others of the present invention are because disclosing of the technology of this paper is conspicuous to those skilled in the art.
Description of drawings
Fig. 1 has shown the pedigree chart of the trichoepithelioma family of an autosomal dominant inheritance.
Fig. 2 is typical patient's a facial photo.
Fig. 3 is patient's a biopsy pathology photo.
Fig. 4 has shown the sequence variation in the CYLD gene, and wherein the 1853rd of SEQ ID NO:1 the disappearance of base has taken place, thereby has caused phase shift mutation, and its encoded protein is shortened.
Embodiment
The inventor is through extensive and deep research, and find first and proved that CYLD and trichoepithelioma are closely related, and found its new function: the change of CYLD will directly cause trichoepithelioma.Finished the present invention on this basis.
By the trichoepithelioma family of an autosomal dominant inheritance being carried out genetic linkage analysis, candidate gene screening and large scale sequencing, confirmed that all patients in this family are in 1853 disappearances that base takes place of No. 10 exon of CYLD gene, thereby caused phase shift mutation, its amino acid coding is shortened, protein translation is unusual, has finally caused the generation of trichoepithelioma.The disappearance of base A if taken place at 1853 places then the total length of only encoding is 496 amino acid whose albumen (SEQ ID NO:5) and the improper little peptide of part in normal CYLD coding 956 amino acid whose albumen of total length (SEQ ID NO:2).
The sudden change of CYLD has caused the human individual to show dermatoma pathology, especially trichoepithelioma.We studies show that, the existence of normal CYLD is the key of skin normal physiological state.
In view of the variation of human CYLD is one of immediate cause that causes trichoepithelioma.Therefore, can be used for diagnosing and treating human trichoepithelioma according to the medicine and the diagnoses and treatment technology of this gene and expression product design thereof.
People CYLD Nucleotide full length sequence of the present invention or its fragment can obtain with the method for pcr amplification method, recombination method or synthetic usually.For the pcr amplification method, can be according to the relevant nucleotide sequence of CYLD, especially open reading frame sequence designs primer, and with commercially available cDNA storehouse or by the prepared cDNA storehouse of ordinary method well known by persons skilled in the art as template, amplification and must relevant sequence.When sequence is longer, usually needs to carry out twice or pcr amplification repeatedly, and then the fragment that each time amplifies is stitched together by proper order.
In case obtained relevant sequence, just can obtain relevant sequence in large quantity with recombination method.This normally is cloned into carrier with it, changes cell again over to, separates obtaining relevant sequence then from the host cell after the propagation by ordinary method.
In addition, also the method for available synthetic is synthesized relevant sequence, especially fragment length more in short-term.Usually, by first synthetic a plurality of small segments, and then connect and to obtain the very long fragment of sequence.
The CYLD encoding sequence is inserted suitable expression vector, change host cell again over to, just can isolate CYLD albumen.
Based on new risk of the present invention, CYLD albumen or polypeptide have many-sided new purposes.These purposes include, but is not limited to: the direct disease (as trichoepithelioma) due to the low or forfeiture and be used to screen the material that promotes the CYLD protein function as pharmacological agent CYLD protein function, and as antibody, polypeptide or other part.The peptide molecule that can stimulate people CYLD protein function that can be used for seeking therapeutic value with the recombinant human CYLD protein screening peptide library of expressing.
On the other hand, the present invention also comprises people CYLD DNA or the polypeptide of its fragment coding has specific polyclonal antibody and monoclonal antibody, especially monoclonal antibody.Here, " specificity " is meant that antibody capable is incorporated into human's CYLD gene product or fragment.Preferably, refer to that those can combine with human's CYLD gene product or fragment but nonrecognition and be incorporated into the antibody of other irrelevant antigen molecule.Among the present invention antibody comprise those can in conjunction with and suppress the proteic molecule of people CYLD, comprise that also those do not influence the antibody of people CYLD protein function.
The present invention not only comprises complete mono-clonal or polyclonal antibody, but also comprises having immunocompetent antibody fragment, as Fab ' or (Fab) 2Fragment; Heavy chain of antibody; Light chain of antibody; Genetically engineered strand Fv molecule; Or chimeric antibody.
Antibody of the present invention can be prepared by the known various technology of those skilled in that art.For example, the human's CYLD gene product of purifying or its have antigenic fragment, can be applied to animal to induce the generation of polyclonal antibody.Similarly, expressing human CYLD albumen or its have antigenic segmental cell and can be used to immune animal and produce antibody.Multiple adjuvant can be used for the enhancing immunity reaction, includes but not limited to freund's adjuvant etc.
Antibody of the present invention also can be monoclonal antibody.This type of monoclonal antibody can utilize hybridoma technology to prepare.Antibody of the present invention comprises the antibody that can block people CYLD protein function and the antibody that does not influence people CYLD protein function.Each antibody-like of the present invention can utilize the fragment or the functional zone of human's CYLD gene product, obtains by the routine immunization technology.These fragments or functional zone can utilize recombinant methods or utilize Peptide synthesizer synthetic.Can come immune animal and produce with the gene product of producing in the prokaryotic cell prokaryocyte (for example E.Coli) with the unmodified form bonded antibody of human's CYLD gene product; With posttranslational modification form bonded antibody (as the albumen or the polypeptide of glycosylation or phosphorylation), can come immune animal and obtain with the gene product that produces in the eukaryotic cell (for example yeast or insect cell).
The proteic antibody of anti-people CYLD can be used in the immunohistochemistry technology, detects the people CYLD albumen in the biopsy specimen.A kind of preferred anti-CYLD antibody is the normal CYLD of nonrecognition but discerns sudden change CYLD (owing to the albumen that the coded albumen of the CYLD of sudden change is more coded than normal CYLD is much smaller, so can design the antibody of only discerning unusual small protein) antibody, perhaps discern normal CYLD but the antibody of nonrecognition sudden change CYLD.Utilize this antibody to the proteic specificity difference of normal and unusual CYLD, the trichoepithelioma susceptibility that can carry out protein level easily detects.
Utilize CYLD albumen of the present invention,, can filter out with CYLD albumen interactional material takes place, as inhibitor, agonist or antagonist etc. by various conventional screening methods.
CYLD albumen of the present invention and antibody thereof, inhibitor, agonist, antagonist etc. when using (administration) in treatment, can provide different effects.Usually, can these materials are formulated in nontoxic, inert and the pharmaceutically acceptable aqueous carrier medium, wherein pH is about 5-8 usually, and preferably pH is about 6-8, although the pH value can be with being changed to some extent by preparation Substance Properties and illness to be treated.The pharmaceutical composition for preparing can carry out administration by conventional route, comprising (but being not limited to): intramuscular, intravenously, subcutaneous or topical (comprising affected area).Be preferably topical.
Normal CYLD polypeptide can be directly used in disease treatment, for example, is used for the treatment of trichoepithelioma aspect.When using CYLD albumen of the present invention, also can use the medicament of other treatment trichoepithelioma simultaneously.
The present invention also provides a kind of pharmaceutical composition, and it contains CYLD albumen of the present invention and the pharmaceutically acceptable carrier or the vehicle of safe and effective amount.This class carrier comprises (but being not limited to): salt solution, damping fluid, glucose, water, glycerine, ethanol and combination thereof.Pharmaceutical preparation should be complementary with administering mode.Pharmaceutical composition of the present invention can be made into the injection form, for example is prepared by ordinary method with the physiological saline or the aqueous solution that contains glucose and other assistant agents.Pharmaceutical composition such as ointment, tablet and capsule can be prepared by ordinary method.Pharmaceutical composition such as ointment, injection, solution, tablet and capsule should be made under aseptic condition.The dosage of activeconstituents is the treatment significant quantity, for example every day about 0.1 microgram/kg body weight-Yue 10 mg/kg body weight.In addition, polypeptide of the present invention also can use with the other treatment agent.
When making pharmaceutical composition, be that the CYLD albumen of safe and effective amount or its antagonist, agonist are applied to Mammals, wherein this safe and effective amount is usually at least about 0.1 microgram/kg body weight, and in most of the cases be no more than about 10 mg/kg body weight, preferably this dosage is about 0.1 microgram/kg body weight-Yue 100 micrograms/kg body weight.Certainly, concrete dosage also should be considered factors such as route of administration, patient health situation, and these all are within the skilled practitioners skill.
The proteic polynucleotide of people CYLD also can be used for multiple therapeutic purpose.Gene therapy technology can be used for treating because cell proliferation, growth or the metabolic disturbance due to the proteic expression of CYLD of the proteic nothing expression of CYLD or unusual/non-activity.The method that structure carries the recombinant viral vector of CYLD gene is found in existing document (Sambrook, et al.).The human's CYLD gene of recombinating in addition can be packaged in the liposome, and then is transferred in the cell.
Polynucleotide imports tissue or intracellular method comprises: directly be injected into polynucleotide in the in-vivo tissue; Or external by carrier (as virus, phage or plasmid etc.) earlier with the polynucleotide transfered cell in, again cell is transplanted in the body etc.
The invention still further relates to the diagnostic testing process of quantitative and detection and localization people CYLD protein level.These tests are known in the art, and comprise that FISH measures and radioimmunoassay.
Whether having the proteic method of CYLD in a kind of detection test sample is to utilize the proteic specific antibody of CYLD to detect, and it comprises: sample is contacted with the CYLD protein specific antibody; Observe whether form antibody complex, formed antibody complex and just represented to exist in the sample CYLD albumen.
The proteic polynucleotide of CYLD can be used for the diagnosis and the treatment of CYLD protein related diseases.Aspect diagnosis, the proteic polynucleotide of CYLD can be used for detecting the proteic expression of CYLD CYLD abnormal exprssion whether or under morbid state.Can be used for the hybridization of biopsy specimen to judge the proteic abnormal expression of CYLD as the CYLD dna sequence dna.Hybridization technique comprises the Southern blotting, Northern blotting, in situ hybridization etc.These technological methods all are disclosed mature technologies, and relevant test kit all can obtain from commercial channels.Part or all of polynucleotide of the present invention can be used as probe stationary on microarray (microarray) or DNA chip (being called " gene chip " again), is used for analyzing the differential expression analysis and the gene diagnosis of tissue gene.Carry out RNA-polymerase chain reaction (RT-PCR) amplification in vitro with the special primer of CYLD albumen and also can detect the proteic transcription product of CYLD.
Whether unusual the present invention also provide a kind of human's CYLD gene that detects to express method, and it comprises step: (a) determine the 1853rd Nucleotide in sequence shown in the SEQ of the human's CYLD gene ID NO:1; (b) whether detection exists base A disappearance in described position.
The sudden change that detects the CYLD gene also can be used for diagnosing trichoepithelioma.Detection can be at cDNA, also can be at genomic dna.The form of CYLD protein mutation comprises that the point mutation compared with normal wild type CYLD dna sequence dna, transposition, disappearance, reorganization and other are any unusual etc.Available existing technology such as Southern blotting, dna sequence analysis, PCR and in situ hybridization detect sudden change.In addition, sudden change might influence proteic expression, therefore can judge indirectly that with Northern blotting, Western blotting gene has or not sudden change.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to people such as normal condition such as Sambrook, molecular cloning: laboratory manual (New York:ColdSpring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.
Embodiment 1
Determining of autosomal dominant trichoepithelioma family
1.1 object
The big family of trichoepithelioma, totally 4 generations, 53 people.Wherein the trichoepithelioma patient has 20 people (Fig. 1).
Embodiment 2
Determine that the CYLD sudden change is the immediate cause that causes trichoepithelioma
2.1 genetic linkage analysis
Utilize microsatellite marker to carry out the linkage analysis of this family.Utilize 384 pairs of micro-satellite primers (RESEARCH GENETICS) that whole genome is scanned altogether, with this family trichoepithelioma assignment of genes gene mapping on people's No. 16 karyomit(e).Further synthetic primer navigates to Disease-causing gene in the genetic distance of 14 between microsatellite marker d16s753 and the d16s3253 (cM) centimorgan, as shown in Figure 1.Nearly 1,000 4 hundred ten thousand base pairs of this physical distance, more than 100 gene.
2.2 candidate gene
With DNA extraction agent box (QIAGEN company) from family's blood sample the extracting genomic dna as template.
To candidate gene, design PCR primer, pcr amplification goes out candidate gene, checks order then.Detect through order-checking, find that the trichoepithelioma of CYLD and this family is closely related candidate gene.
CYLD has been designed 34 pairs of primers to check order.Wherein, when the PCR product that goes out with following a pair of primer amplification checks order, the normal people that the middle CYLD that finds each patient of this trichoepithelioma family contrasts this family finds 1853 the disappearances that base take place of this gene at No. 10 exon, be the 1853rd disappearance that base A has taken place of SEQ ID NO:1, thereby caused phase shift mutation, made its encoded protein shorten to 496 amino acid.This sudden change has finally caused the generation (see figure 4) of trichoepithelioma.
Title Sequence (5 ' → 3 ') Numbering
The CYLD_15 forward tctgcagtga tagcttttct gaca SEQ ID NO：3
CYLD_15 is reverse cagtctcacc aagatgccca atac SEQ ID NO：4
In addition, in order to verify the specificity of this site to the generation of trichoepithelioma, casual inspection 100 situations that do not have the people of close source relation in this site, do not find any sudden change.This illustrates that this site mutation is not due to the polymorphism among the crowd, but the specific mutant site of trichoepithelioma.Therefore shown that CYLD is closely related with the generation of the mankind's trichoepithelioma, this gene and coded product thereof can play an important role to the mankind's trichoepithelioma.
Embodiment 3
The preparation of trichoepithelioma detection kit
Prepare a test kit, it contains:
Title sequence (5 ' → 3 ') numbering concentration
Forward primer tctgcagtga tagcttttct gaca SEQ ID NO:3 dry powder 20D
Reverse primer cagtctcacc aagatgccca atac SEQ ID NO:4 dry powder 20D
The PCR reaction solution contains Taq enzyme dNTP magnesium ion PCR reaction buffer
PCR product purification box contains the solution of PCR product purification in a small amount, DNA adsorption column
Sequencing reaction liquid contains Big Dye
Extract patients'blood 3ml to be detected, use ordinary method (or using specific test kit) from blood, to extract DNA.PCR primer in the trichoepithelioma detection kit is diluted to 1 μ mol/ μ l, is that template is carried out the PCR reaction with the primer that is provided with the DNA that is extracted.Use the PRC product purification box that detection kit provided that the PCR product is carried out purifying.The product of purifying is carried out directly checking order behind the sequencing reaction.Whether the resulting sequence of observation order-checking has phase shift mutation.
Embodiment 4
Preparation of drug combination
Normal protein that CYLD is coded and common ointment material are made the ointment that contains the coded normal protein of CYLD by specific mixed.During use, ointment is imposed on the affected part, CYLD coded normal protein in affected part is replenished, and makes patient owing to the dermatosis that the coded normal protein of shortage CYLD causes is alleviated, until last disappearance.Or the normal protein that CYLD is coded makes injection, during use subcutaneous injection carried out in the affected part, directly replenishes the CYLD proteins encoded of affected area, the physiological situation of patient's skin improved, thereby reach the purpose of treatment.
All quote in this application as a reference at all documents that the present invention mentions, just quoted as a reference separately as each piece document.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Sequence table
＜110〉Shanghai Research Center of Biotechnology
＜120〉utilize human's CYLD gene and coded product thereof to diagnose and treat the method for trichoepithelioma
<130>020613
<160>5
<170>PatentIn version 3.0
<210>1
<211>5371
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<220>
<221>CDS
<222>(392)..(3262)
<400>1
gggggcgggc ccaggtagca ggtttggctg cgcgggggcc gcgcgtcgga gtttccccct 60
ttctagggtg aggatggttc tacacagcca cccggagttc cttagttgaa aggtgcgccc 120
tgctgtgaca gaatgtggta attgtaatct ttaacatttt catgtaaaac atatttcctg 180
atcatctttc cattgtcttc atggaaaatt gataaatatt tgtgccttcc aactctcgtc 240
ttggttgaat gacttcatct taatacaaca tggacaccac gttgctgaaa acatgctttg 300
ggactgccac tgaatttatc ttttgcggtt ttatgacaaa gttattagta gtttcccttt 360
tttgaattag tattttgaag ttaatatcac a atg agt tca ggc tta tgg agc 412
Met Ser Ser Gly Leu Trp Ser
1 5
caa gaa aaa gtc act tca ccc tac tgg gaa gag cgg att ttt tac ttg 460
Gln Glu Lys Val Thr Ser Pro Tyr Trp Glu Glu Arg Ile Phe Tyr Leu
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ctt ctt caa gaa tgc agc gtt aca gac aaa caa aca caa aag ctc ctt 508
Leu Leu Gln Glu Cys Ser Val Thr Asp Lys Gln Thr Gln Lys Leu Leu
25 30 35
aaa gta ccg aag gga agt ata gga cag tat att caa gat cgt tot gtg 556
Lys Val Pro Lys Gly Ser Ile Gly Gln Tyr Ile Gln Asp Arg Ser Val
40 45 50 55
ggg cat tca agg att cct tct gca aaa ggc aag aaa aat cag att gga 604
Gly His Ser Arg Ile Pro Ser Ala Lys Gly Lys Lys Asn Gln Ile Gly
60 65 70
tta aaa att cta gag caa cct cat gca gtt ctc ttt gtt gat gaa aag 652
Leu Lys Ile Leu Glu Gln Pro His Ala Val Leu Phe Val Asp Glu Lys
75 80 85
gat gtt gta gag ata aat gaa aag ttc aca gag tta ctt ttg gca att 700
Asp Val Val Glu Ile Asn Glu Lys Phe Thr Glu Leu Leu Leu Ala Ile
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acc aat tgt gag gag agg ttc agc ctg ttt aaa aac aga aac aga cta 748
Thr Asn Cys Glu Glu Arg Phe Ser Leu Phe Lys Asn Arg Asn Arg Leu
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agt aaa ggc ctc caa ata gac gtg ggc tgt cct gtg aaa gta cag ctg 796
Ser Lys Gly Leu Gln Ile Asp Val Gly Cys Pro Val Lys Val Gln Leu
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aga tct ggg gaa gaa aaa ttt cct gga gtt gta cgc ttc aga gga ccc 844
Arg Ser Gly Glu Glu Lys Phe Pro Gly Val Val Arg Phe Arg Gly Pro
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ctg tta gca gag agg aca gtc tcc gga ata ttc ttt gga gtt gaa ttg 892
Leu Leu Ala Glu Arg Thr Val Ser Gly Ile Phe Phe Gly Val Glu Leu
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ctg gaa gaa ggt cgt ggt caa ggt ttc act gac ggg gtg tac caa ggg 940
Leu Glu Glu Gly Arg Gly Gln Gly Phe Thr Asp Gly Val Tyr Gln Gly
170 175 180
aaa cag ctt ttt cag tgt gat gaa gat tgt ggc gtg ttt gtt gca ttg 988
Lys Gln Leu Phe Gln Cys Asp Glu Asp Cys Gly Val Phe Val Ala Leu
185 190 195
gac aag cta gaa ctc ata gaa gat gat gac act gca ttg gaa agt gat 1036
Asp Lys Leu Glu Leu Ile Glu Asp Asp Asp Thr Ala Leu Glu Ser Asp
200 205 210 215
tac gca ggt cct ggg gac aca atg cag gtc gaa ctt cct cct ttg gaa 1084
Tyr Ala Gly Pro Gly Asp Thr Met Gln Val Glu Leu Pro Pro Leu Glu
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ata aac tcc aga gtt tct ttg aag gtt gga gaa aca ata gaa tct gga 1132
Ile Asn Ser Arg Val Ser Leu Lys Val Gly Glu Thr Ile Glu Ser Gly
235 240 245
aca gtt ata ttc tgt gat gtt ttg cca gga aaa gaa agc tta gga tat 1180
Thr Val Ile Phe Cys Asp Val Leu Pro Gly Lys Glu Ser Leu Gly Tyr
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ttt gtt ggt gtg gac atg gat aac cct att ggc aac tgg gat gga aga 1228
Phe Val Gly Val Asp Met Asp Asn Pro Ile Gly Asn Trp Asp Gly Arg
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ttt gat gga gtg cag ctt tgt agt ttt gcg tgt gtt gaa agt aca att 1276
Phe Asp Gly Val Gln Leu Cys Ser Phe Ala Cys Val Glu Ser Thr Ile
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cta ttg cac atc aat gat atc atc cca gct tta tca gag agt gtg acg 1324
Leu Leu His Ile Asn Asp Ile Ile Pro Ala Leu Ser Glu Ser Val Thr
300 305 310
cag gaa agg agg cct ccc aaa ctt gcc ttt atg tca aga ggt gtt ggg 1372
Gln Glu Arg Arg Pro Pro Lys Leu Ala Phe Met Ser Arg Gly Val Gly
315 320 325
gac aaa ggt tca tcc agt cat aat aaa cca aag gct aca gga tct acc 1420
Asp Lys Gly Ser Ser Ser His Asn Lys Pro Lys Ala Thr Gly Ser Thr
330 335 340
tca gac cct gga aat aga aac aga tct gaa tta ttt tat acc tta aat 1468
Ser Asp Pro Gly Asn Arg Asn Arg Ser Glu Leu Phe Tyr Thr Leu Asn
345 350 355
ggg tct tct gtt gac tca caa cca caa tcc aaa tca aaa aat aca tgg 1516
Gly Ser Ser Val Asp Ser Gln Pro Gln Ser Lys Ser Lys Asn Thr Trp
360 365 370 375
tac att gat gaa gtt gca gaa gac cct gca aaa tct ctt aca gag ata 1564
Tyr Ile Asp Glu Val Ala Glu Asp Pro Ala Lys Ser Leu Thr Glu Ile
380 385 390
tct aca gac ttt gac cgt tct tca cca cca ctc cag cct cct cct gtg 1612
Ser Thr Asp Phe Asp Arg Ser Ser Pro Pro Leu Gln Pro Pro Pro Val
395 400 405
aac tca ctg acc acc gag aac aga ttc cac tct tta cca ttc agt ctc 1660
Asn Ser Leu Thr Thr Glu Asn Arg Phe His Ser Leu Pro Phe Ser Leu
410 415 420
acc aag atg ccc aat acc aat gga agt att ggc cac agt cca ctt tct 1708
Thr Lys Met Pro Asn Thr Asn Gly Ser Ile Gly His Ser Pro Leu Ser
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ctg tca gcc cag tct gta atg gaa gag cta aac act gca ccc gtc caa 1756
Leu Ser Ala Gln Ser Val Met Glu Glu Leu Asn Thr Ala Pro Val Gln
440 445 450 455
gag agt cca ccc ttg gcc atg cct cct ggg aac tca cat ggt cta gaa 1804
Glu Ser Pro Pro Leu Ala Met Pro Pro Gly Asn Ser His Gly Leu Glu
460 465 470
gtg ggc tca ttg gct gaa gtt aag gag aac cct cct ttc tat ggg gta 1852
Val Gly Ser Leu Ala Glu Val Lys Glu Asn Pro Pro Phe Tyr Gly Val
475 480 485
atc cgt tgg atc ggt cag cca cca gga ctg aat gaa gtg ctc gct gga 1900
Ile Arg Trp Ile Gly Gln Pro Pro Gly Leu Asn Glu Val Leu Ala Gly
490 495 500
ctg gaa ctg gaa gat gag tgt gca ggc tgt acg gat gga acc ttc aga 1948
Leu Glu Leu Glu Asp Glu Cys Ala Gly Cys Thr Asp Gly Thr Phe Arg
505 510 515
ggc act cgg tat ttc acc tgt gcc ctg aag aag gcg ctg ttt gtg aaa 1996
Gly Thr Arg Tyr Phe Thr Cys Ala Leu Lys Lys Ala Leu Phe Val Lys
520 525 530 535
ctg aag agc tgc agg cct gac tct agg ttt gca tca ttg cag ccg gtt 2044
Leu Lys Ser Cys Arg Pro Asp Ser Arg Phe Ala Ser Leu Gln Pro Val
540 545 550
tcc aat cag att gag cgc tgt aac tct tta gca ttt gga ggc tac tta 2092
Ser Asn Gln Ile Glu Arg Cys Asn Ser Leu Ala Phe Gly Gly Tyr Leu
555 560 565
agt gaa gta gta gaa gaa aat act cca cca aaa atg gaa aaa gaa ggc 2140
Ser Glu Val Val Glu Glu Asn Thr Pro Pro Lys Met Glu Lys Glu Gly
570 575 580
ttg gag ata atg att ggg aag aag aaa ggc atc cag ggt cat tac aat 2188
Leu Glu Ile Met Ile Gly Lys Lys Lys Gly Ile Gln Gly His Tyr Asn
585 590 595
tct tgt tac tta gac tca acc tta ttc tgc tta ttt gct ttt agt tct 2236
Ser Cys Tyr Leu Asp Ser Thr Leu Phe Cys Leu Phe Ala Phe Ser Ser
600 605 610 615
gtt ctg gac act gtg tta ctt aga ccc aaa gaa aag aac gat gta gaa 2284
Val Leu Asp Thr Val Leu Leu Arg Pro Lys Glu Lys Asn Asp Val Glu
620 625 630
tat tat agt gaa acc caa gag cta ctg agg aca gaa att gtt aat cct 2332
Tyr Tyr Ser Glu Thr Gln Glu Leu Leu Arg Thr Glu Ile Val Asn Pro
635 640 645
ctg aga ata tat gga tat gtg tgt gcc aca aaa att atg aaa ctg agg 2380
Leu Arg Ile Tyr Gly Tyr Val Cys Ala Thr Lys Ile Met Lys Leu Arg
650 655 660
aaa ata ctt gaa aag gtg gag gct gca tca gga ttt acc tct gaa gaa 2428
Lys Ile Leu Glu Lys Val Glu Ala Ala Ser Gly Phe Thr Ser Glu Glu
665 670 675
aaa gat cct gag gaa ttc ttg aat att ctg ttt cat cat att tta agg 2476
Lys Asp Pro Glu Glu Phe Leu Asn Ile Leu Phe His His Ile Leu Arg
680 685 690 695
gta gaa cct ttg cta aaa ata aga tca gca ggt caa aag gta caa gat 2524
Val Glu Pro Leu Leu Lys Ile Arg Ser Ala Gly Gln Lys Val Gln Asp
700 705 710
tgt tac ttc tat caa att ttt atg gaa aaa aat gag aaa gtt ggc gtt 2572
Cys Tyr Phe Tyr Gln Ile Phe Met Glu Lys Asn Glu Lys Val Gly Val
715 720 725
ccc aca att cag cag ttg tta gaa tgg tct ttt atc aac agt aac ctg 2620
Pro Thr Ile Gln Gln Leu Leu Glu Trp Ser Phe Ile Asn Ser Asn Leu
730 735 740
aaa ttt gca gag gca cca tca tgt ctg att att cag atg cct cga ttt 2668
Lys Phe Ala Glu Ala Pro Ser Cys Leu Ile Ile Gln Met Pro Arg Phe
745 750 755
gga aaa gac ttt aaa cta ttt aaa aaa att ttt cct tct ctg gaa tta 2716
Gly Lys Asp Phe Lys Leu Phe Lys Lys Ile Phe Pro Ser Leu Glu Leu
760 765 770 775
aat ata aca gat tta ctt gaa gac act ccc aga cag tgc cgg ata tgt 2764
Asn Ile Thr Asp Leu Leu Glu Asp Thr Pro Arg Gln Cys Arg Ile Cys
780 785 790
gga ggg ctt gca atg tat gag tgt aga gaa tgc tac gac gat ccg gac 2812
Gly Gly Leu Ala Met Tyr Glu Cys Arg Glu Cys Tyr Asp Asp Pro Asp
795 800 805
atc tca gct gga aaa atc aag cag ttt tgt aaa acc tgc aac act caa 2860
Ile Ser Ala Gly Lys Ile Lys Gln Phe Cys Lys Thr Cys Asn Thr Gln
810 815 820
gtc cac ctt cat ccg aag agg ctg aat cat aaa tat aac cca gtg tca 2908
Val His Leu His Pro Lys Arg Leu Asn His Lys Tyr Asn Pro Val Ser
825 830 835
ctt ccc aaa gac tta ccc gac tgg gac tgg aga cac ggc tgc atc cct 2956
Leu Pro Lys Asp Leu Pro Asp Trp Asp Trp Arg His Gly Cys Ile Pro
840 845 850 855
tgc cag aat atg gag tta ttt gct gtt ctc tgc ata gaa aca agc cac 3004
Cys Gln Asn Met Glu Leu Phe Ala Val Leu Cys Ile Glu Thr Ser His
860 865 870
tat gtt gct ttt gtg aag tat ggg aag gac gat tct gcc tgg ctc ttc 3052
Tyr Val Ala Phe Val Lys Tyr Gly Lys Asp Asp Ser Ala Trp Leu Phe
875 880 885
ttt gac agc atg gcc gat cgg gat ggt ggt cag aat ggc ttc aac att 3100
Phe Asp Ser Met Ala Asp Arg Asp Gly Gly Gln Asn Gly Phe Asn Ile
890 895 900
cct caa gtc acc cca tgc cca gaa gta gga gag tac ttg aag atg tct 3148
Pro Gln Val Thr Pro Cys Pro Glu Val Gly Glu Tyr Leu Lys Met Ser
905 910 915
ctg gaa gac ctg cat tcc ttg gac tcc agg aga atc caa ggc tgt gca 3196
Leu Glu Asp Leu His Ser Leu Asp Ser Arg Arg Ile Gln Gly Cys Ala
920 925 930 935
cga aga ctg ctt tgt gat gca tat atg tgc atg tac cag agt cca aca 3244
Arg Arg Leu Leu Cys Asp Ala Tyr Met Cys Met Tyr Gln Ser Pro Thr
940 945 950
atg agt ttg tac aaa taa ctggggtcat cgggaaaggc aaagaaactg 3292
Met Ser Leu Tyr Lys
955
aaggcagagt cctaacgttg catcttattc gagctggcag ttctgttcac gtccattgcc 3352
ggcaatggat gtctttgtgg tgatgatcct tcagaaaagg atgcctctgt ttaaaaacaa 3412
attgcttttg tgtccctgaa gtatttaata agaagcattt tgcactctag aaagtatgtt 3472
tgtgttggtt ttttaagaag tctaaatgaa gttattaata cctgaagctt taagttaagt 3532
gcattgatca tatgatattt ttggaagcat acaattttaa ttgtggaagt ttaaagcctc 3592
ttttagtcca ttgagaatgt aaataaatgt gtcttcttta tggaccaagg atatgaaatc 3652
atttttcttt tgtagctaac ggttgccttg aggaagaaat aatttggttt tattaagagt 3712
ctactctcaa tccagttatt agagatgtac tgagtttgat ttgttaatcc tttctatata 3772
ctgctgatct tgcatgtcta caatctgctc agtttttctg tgtttctgca atagtggtca 3832
gaaaaatact taaattccct taatggtgtt gttttctatt tgttctggtt ttgagataaa 3892
tgagtgattc tgtccccaaa tgtccatttt tgaagtgatt ttcctggagg attagggtat 3952
ttagcagttg aagctcttca ttcatagtag ttactgtcag ctaacaggtt ttttaaggct 4012
tttaactatt aatattttat ggaatggggc aaagtaaatt gatgaaagaa ttggagtgat 4072
aatagtcctt tacaaacata cagtccataa gaaaatgaat ttggcatata gaattattac 4132
aatttcctgg gagagatgga tatttaaacc tctattattt tagacaagac tgtctagaac 4192
ttaagtttga tctgtcagcc agtactccca ttaaattcag tgtagtttca cttgatagaa 4252
tcagatatgt tatcgaaatg ttagcagcag cttcatcctc cttctgatta aagtaagtag 4312
aaatgggatg ttttgtttaa taacagccat agtgtgtgtt tagaccacag cggatgttgt 4372
agaccaggac catagatgat acatgtcagt gctgtggaat gtgcattctc tgagtgttgt 4432
tttgtggtat cattgtcttt cctgaatgac tttctaactg tgcagaaagg cagaaaagtc 4492
atcatatgta tatgtcatat gactttataa aatatttaat gtgacaaaaa gtggaaagaa 4552
tctttacaaa ccctgcaatt acttttttaa aggcactttt actctttggt tttatcattc 4612
cattttgcta atatttacta gctttataaa ttacagtaag gtacaaaaac tcatcttgta 4672
atattttcat ttttgaagtg aaaaagtaca tatattttgc acaaggtttt atactgctaa 4732
gtgcttggtt ggggtggtga gatgatgatt agatcagggg tgaggctgag agactctggg 4792
tttagggcta gccctgcctc catctccctt gggtaaaatg aagggtgtgg ggtaaaagat 4852
gcataaggcc ttttctagct ctgacagcct agaagtccaa tcaccctgta ataaatatgt 4912
gttgaatgaa gaaatgggtg aatgagcttg tcaatgtgat tttaaaaaat tgactacctg 4972
gaggaatgat taggaatcta aatgaagcca gccctcggta tctgcaggtt tctcatccat 5032
ggattcaacc aactgcaaat ggaaaatacg atttttttta aaaaaaggat ggttacatcc 5092
gtattgaaca tgtacagact tttttcttgt cattattctc tgaacaatac aagaactctt 5152
tatgtagcat ttacatttat taggtattat aagtaatcta gagattattt aattaaaata 5212
tacaggagga tgtgtgttta tatgccagaa attctgtacc attttgtatc agggaattga 5272
gcatcttcag atgttggtat ctgcagggat cctggaacca aacccctgca gatactaagg 5332
gctgacgatc taggtaagac tggatttaac agttggaaa 5371
<210>2
<211>956
<212>PRT
＜213〉homo sapiens (Homo sapiens)
<400>2
Met Ser Ser Gly Leu Trp Ser Gln Glu Lys Val Thr Ser Pro Tyr Trp
1 5 10 15
Glu Glu Arg Ile Phe Tyr Leu Leu Leu Gln Glu Cys Ser Val Thr Asp
20 25 30
Lys Gln Thr Gln Lys Leu Leu Lys Val Pro Lys Gly Ser Ile Gly Gln
35 40 45
Tyr Ile Gln Asp Arg Ser Val Gly His Ser Arg Ile Pro Ser Ala Lys
50 55 60
Gly Lys Lys Asn Gln Ile Gly Leu Lys Ile Leu Glu Gln Pro His Ala
65 70 75 80
Val Leu Phe Val Asp Glu Lys Asp Val Val Glu Ile Asn Glu Lys Phe
85 90 95
Thr Glu Leu Leu Leu Ala Ile Thr Asn Cys Glu Glu Arg Phe Ser Leu
100 105 110
Phe Lys Asn Arg Asn Arg Leu Ser Lys Gly Leu Gln Ile Asp Val Gly
115 120 125
Cys Pro Val Lys Val Gln Leu Arg Ser Gly Glu Glu Lys Phe Pro Gly
130 135 140
Val Val Arg Phe Arg Gly Pro Leu Leu Ala Glu Arg Thr Val Ser Gly
145 150 155 160
Ile Phe Phe Gly Val Glu Leu Leu Glu Glu Gly Arg Gly Gln Gly Phe
165 170 175
Thr Asp Gly Val Tyr Gln Gly Lys Gln Leu Phe Gln Cys Asp Glu Asp
180 185 190
Cys Gly Val Phe Val Ala Leu Asp Lys Leu Glu Leu Ile Glu Asp Asp
195 200 205
Asp Thr Ala Leu Glu Ser Asp Tyr Ala Gly Pro Gly Asp Thr Met Gln
210 215 220
Val Glu Leu Pro Pro Leu Glu Ile Asn Ser Arg Val Ser Leu Lys Val
225 230 235 240
Gly Glu Thr Ile Glu Ser Gly Thr Val Ile Phe Cys Asp Val Leu Pro
245 250 255
Gly Lys Glu Ser Leu Gly Tyr Phe Val Gly Val Asp Met Asp Asn Pro
260 265 270
Ile Gly Asn Trp Asp Gly Arg Phe Asp Gly Val Gln Leu Cys Ser Phe
275 280 285
Ala Cys Val Glu Ser Thr Ile Leu Leu His Ile Asn Asp Ile Ile Pro
290 295 300
Ala Leu Ser Glu Ser Val Thr Gln Glu Arg Arg Pro Pro Lys Leu Ala
305 310 315 320
Phe Met Ser Arg Gly Val Gly Asp Lys Gly Ser Ser Ser His Asn Lys
325 330 335
Pro Lys Ala Thr Gly Ser Thr Ser Asp Pro Gly Asn Arg Asn Arg Ser
340 345 350
Glu Leu Phe Tyr Thr Leu Asn Gly Ser Ser Val Asp Ser Gln Pro Gln
355 360 365
Ser Lys Ser Lys Asn Thr Trp Tyr Ile Asp Glu Val Ala Glu Asp Pro
370 375 380
Ala Lys Ser Leu Thr Glu Ile Ser Thr Asp Phe Asp Arg Ser Ser Pro
385 390 395 400
Pro Leu Gln Pro Pro Pro Val Asn Ser Leu Thr Thr Glu Asn Arg Phe
405 410 415
His Ser Leu Pro Phe Ser Leu Thr Lys Met Pro Asn Thr Asn Gly Ser
420 425 430
Ile Gly His Ser Pro Leu Ser Leu Ser Ala Gln Ser Val Met Glu Glu
435 440 445
Leu Asn Thr Ala Pro Val Gln Glu Ser Pro Pro Leu Ala Met Pro Pro
450 455 460
Gly Asn Ser His Gly Leu Glu Val Gly Ser Leu Ala Glu Val Lys Glu
465 470 475 480
Asn Pro Pro Phe Tyr Gly Val Ile Arg Trp Ile Gly Gln Pro Pro Gly
485 490 495
Leu Asn Glu Val Leu Ala Gly Leu Glu Leu Glu Asp Glu Cys Ala Gly
500 505 510
Cys Thr Asp Gly Thr Phe Arg Gly Thr Arg Tyr Phe Thr Cys Ala Leu
515 520 525
Lys Lys Ala Leu Phe Val Lys Leu Lys Ser Cys Arg Pro Asp Ser Arg
530 535 540
Phe Ala Ser Leu Gln Pro Val Ser Asn Gln Ile Glu Arg Cys Asn Ser
545 550 555 560
Leu Ala Phe Gly Gly Tyr Leu Ser Glu Val Val Glu Glu Asn Thr Pro
565 570 575
Pro Lys Met Glu Lys Glu Gly Leu Glu Ile Met Ile Gly Lys Lys Lys
580 585 590
Gly Ile Gln Gly His Tyr Asn Ser Cys Tyr Leu Asp Ser Thr Leu Phe
595 600 605
Cys Leu Phe Ala Phe Ser Ser Val Leu Asp Thr Val Leu Leu Arg Pro
610 615 620
Lys Glu Lys Asn Asp Val Glu Tyr Tyr Ser Glu Thr Gln Glu Leu Leu
625 630 635 640
Arg Thr Glu Ile Val Asn Pro Leu Arg Ile Tyr Gly Tyr Val Cys Ala
645 650 655
Thr Lys Ile Met Lys Leu Arg Lys Ile Leu Glu Lys Val Glu Ala Ala
660 665 670
Ser Gly Phe Thr Ser Glu Glu Lys Asp Pro Glu Glu Phe Leu Asn Ile
675 680 685
Leu Phe His His Ile Leu Arg Val Glu Pro Leu Leu Lys Ile Arg Ser
690 695 700
Ala Gly Gln Lys Val Gln Asp Cys Tyr Phe Tyr Gln Ile Phe Met Glu
705 710 715 720
Lys Asn Glu Lys Val Gly Val Pro Thr Ile Gln Gln Leu Leu Glu Trp
725 730 735
Ser Phe Ile Asn Ser Asn Leu Lys Phe Ala Glu Ala Pro Ser Cys Leu
740 745 750
Ile Ile Gln Met Pro Arg Phe Gly Lys Asp Phe Lys Leu Phe Lys Lys
755 760 765
Ile Phe Pro Ser Leu Glu Leu Asn Ile Thr Asp Leu Leu Glu Asp Thr
770 775 780
Pro Arg Gln Cys Arg Ile Cys Gly Gly Leu Ala Met Tyr Glu Cys Arg
785 790 795 800
Glu Cys Tyr Asp Asp Pro Asp Ile Ser Ala Gly Lys Ile Lys Gln Phe
805 810 815
Cys Lys Thr Cys Asn Thr Gln Val His Leu His Pro Lys Arg Leu Asn
820 825 830
His Lys Tyr Asn Pro Val Ser Leu Pro Lys Asp Leu Pro Asp Trp Asp
835 840 845
Trp Arg His Gly Cys Ile Pro Cys Gln Asn Met Glu Leu Phe Ala Val
850 855 860
Leu Cys Ile Glu Thr Ser His Tyr Val Ala Phe Val Lys Tyr Gly Lys
865 870 875 880
Asp Asp Ser Ala Trp Leu Phe Phe Asp Ser Met Ala Asp Arg Asp Gly
885 890 895
Gly Gln Asn Gly Phe Asn Ile Pro Gln Val Thr Pro Cys Pro Glu Val
900 905 910
Gly Glu Tyr Leu Lys Met Ser Leu Glu Asp Leu His Ser Leu Asp Ser
915 920 925
Arg Arg Ile Gln Gly Cys Ala Arg Arg Leu Leu Cys Asp Ala Tyr Met
930 935 940
Cys Met Tyr Gln Ser Pro Thr Met Ser Leu Tyr Lys
945 950 955
<210>3
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>3
tctgcagtga tagcttttct gaca 24
<210>4
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>4
cagtctcacc aagatgccca atac 24
<210>5
<211>496
<212>PRT
＜213〉homo sapiens (Homo sapiens)
<400>5
Met Ser Ser Gly Leu Trp Ser Gln Glu Lys Val Thr Ser Pro Tyr Trp
1 5 10 15
Glu Glu Arg Ile Phe Tyr Leu Leu Leu Gln Glu Cys Ser Val Thr Asp
20 25 30
Lys Gln Thr Gln Lys Leu Leu Lys Val Pro Lys Gly Ser Ile Gly Gln
35 40 45
Tyr Ile Gln Asp Arg Ser Val Gly His Ser Arg Ile Pro Ser Ala Lys
50 55 60
Gly Lys Lys Asn Gln Ile Gly Leu Lys Ile Leu Glu Gln Pro His Ala
65 70 75 80
Val Leu Phe Val Asp Glu Lys Asp Val Val Glu Ile Asn Glu Lys Phe
85 90 95
Thr Glu Leu Leu Leu Ala Ile Thr Asn Cys Glu Glu Arg Phe Ser Leu
100 105 110
Phe Lys Asn Arg Asn Arg Leu Ser Lys Gly Leu Gln Ile Asp Val Gly
115 120 125
Cys Pro Val Lys Val Gln Leu Arg Ser Gly Glu Glu Lys Phe Pro Gly
130 135 140
Val Val Arg Phe Arg Gly Pro Leu Leu Ala Glu Arg Thr Val Ser Gly
145 150 155 160
Ile Phe Phe Gly Val Glu Leu Leu Glu Glu Gly Arg Gly Gln Gly Phe
165 170 175
Thr Asp Gly Val Tyr Gln Gly Lys Gln Leu Phe Gln Cys Asp Glu Asp
180 185 190
Cys Gly Val Phe Val Ala Leu Asp Lys Leu Glu Leu Ile Glu Asp Asp
195 200 205
Asp Thr Ala Leu Glu Ser Asp Tyr Ala Gly Pro Gly Asp Thr Met Gln
210 215 220
Val Glu Leu Pro Pro Leu Glu Ile Asn Ser Arg Val Ser Leu Lys Val
225 230 235 240
Gly Glu Thr Ile Glu Ser Gly Thr Val Ile Phe Cys Asp Val Leu Pro
245 250 255
Gly Lys Glu Ser Leu Gly Tyr Phe Val Gly Val Asp Met Asp Asn Pro
260 265 270
Ile Gly Asn Trp Asp Gly Arg Phe Asp Gly Val Gln Leu Cys Ser Phe
275 280 285
Ala Cys Val Glu Ser Thr Ile Leu Leu His Ile Asn Asp Ile Ile Pro
290 295 300
Ala Leu Ser Glu Ser Val Thr Gln Glu Arg Arg Pro Pro Lys Leu Ala
305 310 315 320
Phe Met Ser Arg Gly Val Gly Asp Lys Gly Ser Ser Ser His Asn Lys
325 330 335
Pro Lys Ala Thr Gly Ser Thr Ser Asp Pro Gly Asn Arg Asn Arg Ser
340 345 350
Glu Leu Phe Tyr Thr Leu Asn Gly Ser Ser Val Asp Ser Gln Pro Gln
355 360 365
Ser Lys Ser Lys Asn Thr Trp Tyr Ile Asp Glu Val Ala Glu Asp Pro
370 375 380
Ala Lys Ser Leu Thr Glu Ile Ser Thr Asp Phe Asp Arg Ser Ser Pro
385 390 395 400
Pro Leu Gln Pro Pro Pro Val Asn Ser Leu Thr Thr Glu Asn Arg Phe
405 410 415
His Ser Leu Pro Phe Ser Leu Thr Lys Met Pro Asn Thr Asn Gly Ser
420 425 430
Ile Gly His Ser Pro Leu Ser Leu Ser Ala Gln Ser Val Met Glu Glu
435 440 445
Leu Asn Thr Ala Pro Val Gln Glu Ser Pro Pro Leu Ala Met Pro Pro
450 455 460
Gly Asn Ser His Gly Leu Glu Val Gly Ser Leu Ala Glu Val Lys Glu
465 470 475 480
Asn Pro Pro Phe Tyr Gly Val Ser Val Gly Ser Val Ser His Gln Asp
485 490 495

Claims (4)

1. the CYLD gene purposes in the reagent of preparation diagnosis trichoepithelioma susceptibility, the nucleotide sequence of wherein said CYLD gene is the 1853rd A disappearance among sequence shown in the SEQ ID NO:1 and the SEQ ID NO:1.
2. the CYLD albumen purposes in the reagent of preparation diagnosis trichoepithelioma, the proteic aminoacid sequence of wherein said CYLD is the aminoacid sequence shown in the SEQ ID NO:5.
3. test kit that detects trichoepithelioma, it is characterized in that, it comprises the primer of specific amplification CYLD gene or transcript, the PCR reaction solution, PCR product purification box, and sequencing reaction liquid, wherein said PCR product purification box contains solution and the DNA adsorption column that is useful on the PCR product purification, and described test kit also contains the reagent that is selected from down group:
(a) with mutable site bonded probe;
(b) restriction enzyme in identification mutational site;
Wherein said mutable site or mutational site are the 1853rd A disappearances among the SEQ ID NO:1.
4. test kit as claimed in claim 3 is characterized in that, described primer is that sequence is the primer of SEQ ID NO:3 and 4.
CNB021108579A 2002-02-10 2002-02-10 Method for diagnosis and treatment of hair eipidermal tumor using human's CYLD gene and its coded products Expired - Lifetime CN100436595C (en)

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Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Loss of heterozygosity at cylindromatosis gene locus, CYLD,in sporadic skin adnexal tumours. N Leonard , et al.J CLIN PATHOL,Vol.54 No.9. 2001
Loss of heterozygosity at cylindromatosis gene locus, CYLD,in sporadic skin adnexal tumours. N Leonard , et al.J CLIN PATHOL,Vol.54 No.9. 2001 *

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