CN100478354C - Novel human protein with cancer inhibiting function and its code sequence - Google Patents

Novel human protein with cancer inhibiting function and its code sequence Download PDF

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CN100478354C
CN100478354C CNB021364001A CN02136400A CN100478354C CN 100478354 C CN100478354 C CN 100478354C CN B021364001 A CNB021364001 A CN B021364001A CN 02136400 A CN02136400 A CN 02136400A CN 100478354 C CN100478354 C CN 100478354C
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CN1473849A (en
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顾健人
杨胜利
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SHANGHAI XINSHIJIE GENE TECHN DEVELOPMENT Co Ltd
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SHANGHAI XINSHIJIE GENE TECHN DEVELOPMENT Co Ltd
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Priority to PCT/CN2003/000639 priority patent/WO2004014946A1/en
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Abstract

The present invention discloses a new kind of human protein with cancer inhibiting function, polynucleotides encoding this polypeptide and recombination process to produce the polypeptide. The present invention also discloses the method of applying the polypeptide in treating various diseases, such as cancer. The present invention also discloses the agonist resisting the polypeptide and its treatment effect. The present invention also discloses the application of the polynucleotides encoding this human protein with cancer inhibiting function.

Description

New people's albumen and encoding sequence thereof with cancer suppressing function
Technical field
The invention belongs to biological technical field, specifically, the present invention relates to new coding and have the proteic polynucleotide of people of cancer suppressing function and the polypeptide of this polynucleotide encoding.The invention still further relates to the purposes and the preparation of these polynucleotide and polypeptide.
Background technology
The research of people's gene group is international focus at present, removes human chromosome DNA large scale sequencing, outside the method for expressed sequence order-checking (EST), also lacks the screening that begins from function and has the high-throughout method of functional gene.
Cancer is one of principal disease of harm humans health.In order to treat effectively and prophylaxis of tumours, people more and more pay close attention to genetic treatment of tumor at present.Therefore, this area presses for people's albumen and the agonist/inhibitor thereof that development research has cancer suppressing function.
Summary of the invention
The purpose of this invention is to provide the new people's protein polypeptide of a class with cancer suppressing function with and fragment, analogue and derivative.
Another object of the present invention provides the polynucleotide of these polypeptide of coding.
Another object of the present invention provides the method for these polypeptide of production and the purposes of this polypeptide and encoding sequence.
In a first aspect of the present invention, novel isolated protein polypeptide with cancer suppressing function is provided, it comprises the polypeptide of the aminoacid sequence with the group of being selected from down: SEQ ID NO:3,6,9,12,15,18; Or its conservative property variation polypeptide or its active fragments or its reactive derivative.
Preferably, this polypeptide is the polypeptide with aminoacid sequence of the group of being selected from down: SEQ ID NO:3,6,9,12,15,18.
In a second aspect of the present invention, a kind of isolating polynucleotide are provided, it comprises a nucleotide sequence, and this nucleotide sequence is shown at least 85% homogeny with a kind of nucleotides sequence that is selected from down group: the polynucleotide of the above-mentioned protein polypeptide with cancer suppressing function of (a) encoding; (b) with polynucleotide (a) complementary polynucleotide.Preferably, the polypeptide of this polynucleotide encoding has the aminoacid sequence of the group of being selected from down: SEQ ID NO:3,6,9,12,15,18.More preferably, the sequence of these polynucleotide is selected from down group: SEQ ID NO:2,5,8,11,14,17 coding region sequence or full length sequence.
In a third aspect of the present invention, the carrier that contains above-mentioned polynucleotide is provided, and has been transformed or host cell of transduceing or the host cell that is directly transformed or transduce by above-mentioned polynucleotide by this carrier.
In a fourth aspect of the present invention, the preparation method who prepares the polypeptide of the protein-active with cancer suppressing function is provided, this method comprises: (a) have under the proteic condition of cancer suppressing function suitable the expression, cultivate the above-mentioned host cell that is transformed or transduce; (b) from culture, isolate the polypeptide of protein-active with cancer suppressing function.
In a fifth aspect of the present invention, provide and above-mentioned protein polypeptide specificity bonded antibody with cancer suppressing function.The nucleic acid molecule that can be used for detecting also is provided, and it contains, and continuous 10 Nucleotide are to full length nucleotide in the above-mentioned polynucleotide, and preferably it contains the about 15-1000 of a successive Nucleotide.
In a sixth aspect of the present invention, a kind of pharmaceutical composition is provided, it contains the protein polypeptide and the pharmaceutically acceptable carrier with cancer suppressing function of the present invention of safe and effective amount.These pharmaceutical compositions can be treated illnesss such as cancer and cellular abnormality propagation.
Others of the present invention are because the disclosure of this paper is conspicuous to those skilled in the art.
Embodiment
The present invention adopts large-scale cDNA clone transfection cancer cells, has on the basis of cancer suppressing action in acquisition, proves new gene through order-checking, further obtains full length cDNA clone.DNA transfection evidence, the albumen with cancer suppressing function of the present invention has the effect that suppresses clone's formation, its inhibiting rate 〉=50% to liver cancer cell 7721.
As used herein, " isolating " is meant that material separates (if natural substance, primal environment promptly is a natural surroundings) from its primal environment.Do not have separation and purification as polynucleotide under the native state in the active somatic cell and polypeptide, but same polynucleotide or polypeptide as from native state with in other materials that exist separately, then for separation and purification.
As used herein, " isolating albumen or polypeptide with cancer suppressing function " is meant that the protein polypeptide with cancer suppressing function is substantially free of natural relative other albumen, lipid, carbohydrate or other material.Those skilled in the art can have the albumen of cancer suppressing function with the purified technology of protein purifying of standard.Basically pure polypeptide can produce single master tape on non-reduced polyacrylamide gel.
Polypeptide of the present invention can be recombinant polypeptide, natural polypeptides, synthetic polypeptide, preferred recombinant polypeptide.Polypeptide of the present invention can be the product of natural purifying, or the product of chemosynthesis, or uses recombinant technology to produce from protokaryon or eucaryon host (for example, bacterium, yeast, higher plant, insect and mammalian cell).The host used according to the recombinant production scheme, polypeptide of the present invention can be glycosylated, maybe can be nonglycosylated.Polypeptide of the present invention also can comprise or not comprise initial methionine residues.
The present invention also comprises the proteic fragment of the people with cancer suppressing function, derivative and analogue.As used herein, term " fragment ", " derivative " are meant with " analogue " and keep natural identical biological function or the active polypeptide of people's albumen with cancer suppressing function of the present invention basically.Polypeptide fragment of the present invention, derivative or analogue can be that (i) has one or more conservative or substituted polypeptide of non-conservation amino-acid residue (preferred conservative amino acid residue), and the amino-acid residue of such replacement can be also can not encoded by genetic code, or (ii) in one or more amino-acid residues, has a polypeptide of substituted radical, or (iii) mature polypeptide and another compound (such as the compound that prolongs the polypeptide transformation period, polyoxyethylene glycol for example) merge formed polypeptide, or (iv) additional aminoacid sequence is fused to this peptide sequence and the polypeptide that forms (as leader sequence or secretion sequence or be used for the sequence or the proteinogen sequence of this polypeptide of purifying).According to the instruction of this paper, these fragments, derivative and analogue belong to the known scope of those skilled in the art.
Polynucleotide of the present invention can be dna form or rna form.Dna form comprises the DNA of cDNA, genomic dna or synthetic.DNA can be strand or double-stranded.DNA can be coding strand or noncoding strand.Be example with FP17388 albumen (in this application, its clone numbering is adopted in proteinic name), the coding region sequence of encoding mature polypeptide can be identical with the coding region sequence shown in the SEQ ID NO:2 or the varient of degeneracy.As used herein, " varient of degeneracy " is meant that for FP17388 coding has the protein of SEQ ID NO:3, but with the differentiated nucleotide sequence of coding region sequence shown in the SEQ ID NO:2.Be example with FP17814 albumen again, the coding region sequence of encoding mature polypeptide can be identical with the coding region sequence shown in the SEQ ID NO:5 or the varient of degeneracy; " varient of degeneracy " is meant that for FP17814 coding has the protein of SEQ ID NO:6, but with the differentiated nucleotide sequence of coding region sequence shown in the SEQ ID NO:5.Have the albumen of cancer suppressing function for of the present invention other, can the rest may be inferred.
The polynucleotide of encoding mature polypeptide comprise: the encoding sequence of an encoding mature polypeptide; The encoding sequence of mature polypeptide and various additional code sequence; Encoding sequence of mature polypeptide (with optional additional code sequence) and non-coding sequence.
Term " polynucleotide of coded polypeptide " can be the polynucleotide that comprise this polypeptide of encoding, and also can be the polynucleotide that also comprise additional code and/or non-coding sequence.
The invention still further relates to the varient of above-mentioned polynucleotide, its coding has the polypeptide of identical aminoacid sequence or fragment, analogue and the derivative of polypeptide with the present invention.The varient of these polynucleotide can be the allelic variant of natural generation or the varient that non-natural takes place.These nucleotide diversity bodies comprise and replace varient, deletion mutation body and insert varient.As known in the art, allelic variant is the replacement form of polynucleotide, and it may be replacement, disappearance or the insertion of one or more Nucleotide, but can be from not changing the function of its encoded polypeptides in fact.
The invention still further relates to and above-mentioned sequence hybridization and two sequences between have at least 50%, preferably at least 70%, the polynucleotide of at least 80% homogeny more preferably.The present invention be more particularly directed under stringent condition and the interfertile polynucleotide of polynucleotide of the present invention.In the present invention, " stringent condition " is meant: (1) than hybridization under low ionic strength and the comparatively high temps and wash-out, as 0.2 * SSC, and 0.1%SDS, 60 ℃; Or (2) hybridization the time is added with denaturing agent, as 50% (v/v) methane amide, 0.1% calf serum/0.1%Ficoll, 42 ℃ etc.; Or (3) only at the homogeny between the two sequences at least more than 95%, be more preferably 97% and just hybridize when above.And the polypeptide of interfertile polynucleotide encoding has identical biological function (is example with FP17388 albumen) and activity with the mature polypeptide shown in the SEQ IDNO:3.
The invention still further relates to nucleic acid fragment with above-mentioned sequence hybridization.As used herein, the length of " nucleic acid fragment " contains 15 Nucleotide at least, better is at least 30 Nucleotide, is more preferably at least 50 Nucleotide, preferably more than at least 100 Nucleotide.The amplification technique (as PCR) that nucleic acid fragment can be used for nucleic acid has the proteic polynucleotide of cancer suppressing function to determine and/or to separate to encode.
Polypeptide among the present invention and polynucleotide preferably provide with isolating form, more preferably are purified to homogeneous.
Dna sequence dna of the present invention can obtain with several method.For example, with hybridization technique DNA isolation well known in the art.These technology including, but not limited to: 1) with probe and genome or the hybridization of cDNA library to detect homology nucleotide sequence and 2) antibody screening of expression library to be to detect the dna fragmentation of the clone with common structure feature.
The proteic specific DNA fragment sequence that coding has cancer suppressing function produces also and can obtain with following method: 1) separate double chain DNA sequence from genomic dna; 2) the chemical synthesising DNA sequence is to obtain the double-stranded DNA of required polypeptide.
When the whole aminoacid sequence of the polypeptide product of needs was known, the direct chemical of dna sequence dna is synthetic to be the method for often selecting for use.When if required amino acid whose whole sequence is not known, the direct chemical of dna sequence dna is synthetic to be impossible, and the method for selecting for use is the separation of cDNA sequence.The standard method that separates interested cDNA is from the donorcells separating mRNA of this gene of high expression level and carries out reverse transcription, forms plasmid or phage cDNA library.Extract the existing multiple proven technique of method of mRNA, test kit also can obtain (Qiagene) from commercial channels.And the construction cDNA library also is usual method (Sambrook, et al., Molecular Cloning, A Laboratory Manual, Cold SpringHarbor Laboratory.New York, 1989).Also can obtain the cDNA library of commercial offers, as the different cDNA library of Clontech company.When being used in combination the polymeric enzyme reaction technology, even few expression product also can be cloned.
Available ordinary method is screened gene of the present invention from these cDNA libraries.These methods include, but is not limited to: (1) DNA-DNA or DNA-RNA hybridization; (2) function of marker gene occurs or forfeiture; (3) mensuration has the level of the proteic transcript of cancer suppressing function; (4), detect the protein product of genetic expression by immunological technique or mensuration biologic activity.Aforesaid method can singly be used, but also several different methods combined utilization.
In (1) kind method, hybridizing used probe is and any a part of homology of polynucleotide of the present invention that at least 15 Nucleotide of its length better are at least 30 Nucleotide, are more preferably at least 50 Nucleotide, preferably at least 100 Nucleotide.In addition, the length of probe within 2kb, preferably is within the 1kb usually.Probe used herein is the dna sequence dna of chemosynthesis on the basis of gene DNA sequence information of the present invention normally.Gene of the present invention itself or fragment are certainly as probe.The mark of dna probe can be used radio isotope, fluorescein or enzyme (as alkaline phosphatase) etc.
In (4) kind method, detect the protein product of protein gene expression and can use immunological technique such as Western blotting, radioimmunoprecipitation, enzyme-linked immunosorbent assay (ELISA) etc. with cancer suppressing function.
Use method (Saiki, the et al.Science 1985 of round pcr DNA amplification/RNA; 230:1350-1354) be optimized for acquisition gene of the present invention.When particularly being difficult to obtain the cDNA of total length from the library, can preferably use RACE method (the terminal rapid amplifying method of RACE-cDNA), the primer that is used for PCR can suitably be selected according to sequence information of the present invention disclosed herein, and available ordinary method is synthetic.Available ordinary method is as the DNA/RNA fragment by gel electrophoresis separation and purifying amplification.
The gene of the present invention that obtains as mentioned above, perhaps the available ordinary method of mensuration of the nucleotide sequence of various dna fragmentations etc. such as dideoxy chain termination (Sanger et al.PNAS, 1977,74:5463-5467).This class nucleotide sequencing is available commercial sequencing kit etc. also.In order to obtain the cDNA sequence of total length, order-checking need be carried out repeatedly.Sometimes need to measure a plurality of clones' cDNA sequence, just can be spliced into the cDNA sequence of total length.
The present invention also relates to comprise the carrier of polynucleotide of the present invention, and the host cell that produces through genetically engineered with carrier of the present invention or albumen coded sequence with cancer suppressing function, and the method that produces polypeptide of the present invention through recombinant technology.
By the recombinant DNA technology of routine, can utilize polymerized nucleoside acid sequence of the present invention to can be used to express or produce the protein polypeptide with cancer suppressing function of reorganization.In general following steps are arranged:
(1). have the proteic polynucleotide of people (or varient) of cancer suppressing function with coding of the present invention, or transform or the transduction proper host cell with the recombinant expression vector that contains these polynucleotide;
(2). the host cell of in suitable medium, cultivating;
(3). separation, protein purification from substratum or cell.
Among the present invention, the people's albumen polynucleotide sequence with cancer suppressing function can be inserted in the recombinant expression vector.Term " recombinant expression vector " refers to that bacterial plasmid well known in the art, phage, yeast plasmid, vegetable cell virus, mammalian cell virus are as adenovirus, retrovirus or other carriers.The carrier of Shi Yonging includes but not limited in the present invention: the expression vector based on T7 of expressing in bacterium; The pMSXND expression vector of in mammalian cell, expressing and at the carrier that derives from baculovirus of expressed in insect cells.In a word, as long as can duplicate in host and stablize, any plasmid and carrier can be used.A key character of expression vector is to contain replication orgin, promotor, marker gene and translation controlling elements usually.
Method well-known to those having ordinary skill in the art can be used to make up and contains people's encoding histone dna sequence dna with cancer suppressing function and suitable transcribing/the translate expression vector of control signal.These methods comprise extracorporeal recombinant DNA technology, DNA synthetic technology, the interior recombinant technology of body etc.Described dna sequence dna can effectively be connected on the suitable promotor in the expression vector, and is synthetic to instruct mRNA.The representative example of these promotors has: colibacillary lac or trp promotor; Lambda particles phage P LPromotor; Eukaryotic promoter comprises LTRs and some other known may command gene expression promoter in protokaryon or eukaryotic cell or its virus of CMV immediate early promoter, early stage and late period SV40 promotor, retrovirus.Expression vector also comprises ribosome bind site and the transcription terminator that translation initiation is used.
In addition, expression vector preferably comprises one or more selected markers, to be provided for selecting the phenotypic character of transformed host cells, cultivate Tetrahydrofolate dehydrogenase, neomycin resistance and the green fluorescent protein (GFP) of usefulness as eukaryotic cell, or be used for colibacillary tsiklomitsin or amicillin resistance.
Comprise the carrier of above-mentioned suitable dna sequence dna and suitable promotor or control sequence, can be used to transform appropriate host cell, so that it can marking protein.
Host cell can be a prokaryotic cell prokaryocyte, as bacterial cell; Or eukaryotic cell such as low, as yeast cell; Or higher eucaryotic cells, as mammalian cell.Representative example has: intestinal bacteria, streptomyces; The bacterial cell of Salmonella typhimurium; Fungal cell such as yeast; Vegetable cell; The insect cell of fruit bat S2 or Sf9; The zooblast of CHO, COS or Bowes melanoma cells etc.
When polynucleotide of the present invention are expressed in higher eucaryotic cells, be enhanced if will make to transcribe when in carrier, inserting enhancer sequence.Enhanser is the cis acting factor of DNA, and nearly 10 to 300 base pairs act on promotor transcribing with enhancing gene usually.Can for example be included in the SV40 enhanser of 100 to 270 base pairs of replication origin side in late period one, at the polyoma enhanser of replication origin side in late period one and adenovirus enhanser etc.
Persons skilled in the art all know how to select appropriate carriers, promotor, enhanser and host cell.
Can carry out with routine techniques well known to those skilled in the art with the recombinant DNA transformed host cell.When the host was prokaryotic organism such as intestinal bacteria, the competent cell that can absorb DNA can be used CaCl in exponential growth after date results 2Method is handled, and used step is well-known in this area.Alternative is to use MgCl 2If desired, transforming also the method for available electroporation carries out.When the host is an eukaryote, can select following DNA transfection method for use: coprecipitation of calcium phosphate method, conventional mechanical method such as microinjection, electroporation, liposome packing etc.
The transformant that obtains can be cultivated with ordinary method, expresses the polypeptide of coded by said gene of the present invention.According to used host cell, used substratum can be selected from various conventional substratum in the cultivation.Under the condition that is suitable for the host cell growth, cultivate.After host cell grows into suitable cell density, induce the promotor of selection with suitable method (as temperature transition or chemical induction), cell is cultivated for some time again.
Recombinant polypeptide in the above methods can wrap by in cell, extracellular or on cytolemma, express or be secreted into the extracellular.If desired, can utilize its physics, the separating by various separation methods with other characteristic and the albumen of purification of Recombinant of chemistry.These methods are well-known to those skilled in the art.The example of these methods includes, but are not limited to: conventional renaturation handles, with protein precipitant handle (salt analysis method), centrifugal, the broken bacterium of infiltration, superly handle, the combination of super centrifugal, sieve chromatography (gel-filtration), adsorption chromatography, ion exchange chromatography, high performance liquid chromatography (HPLC) and other various liquid chromatography (LC) technology and these methods.
The people's albumen or the polypeptide with cancer suppressing function of reorganization are of use in many ways.These purposes include, but is not limited to: directly have the disease due to the low or forfeiture of the protein function of cancer suppressing function as pharmacological agent and be used to screen and promote or antagonism has antibody, polypeptide or other part of the protein function of cancer suppressing function.For example, antibody can be used for activating or suppressing to have the proteic function of people of cancer suppressing function.The people's protein screening peptide library that has a cancer suppressing function with the reorganization of expressing can be used for seeking the peptide molecule that can suppress or stimulate the people's protein function with cancer suppressing function of therapeutic value.
The present invention also provides screening of medicaments to improve (agonist) or check the method that (antagonist) has the proteic medicament of people of cancer suppressing function to identify.Agonist improves the biological function such as stimulate cellular proliferation of the people's albumen with cancer suppressing function, and antagonist prevention disorder such as the various cancer relevant with cell hyperproliferation with treatment.For example, can be in the presence of medicine, the proteic film preparation of people that mammalian cell or expression is had cancer suppressing function is cultivated with the people's albumen with cancer suppressing function of mark.Measure the medicine raising then or check this interactional ability.
The proteic antagonist of people with cancer suppressing function comprises antibody, compound, acceptor disappearance thing and the analogue etc. that filter out.The proteic antagonist of people with cancer suppressing function can and be eliminated its function with the people's protein binding with cancer suppressing function, or suppresses to have the proteic generation of people of cancer suppressing function, or combines with the avtive spot of polypeptide and to make polypeptide can not bring into play biological function.The proteic antagonist of people with cancer suppressing function can be used for therepic use.
In screening during as the compound of antagonist, albumen of the present invention can be added during bioanalysis measures, determine by measuring albumen and the interaction between its acceptor that compounds affect has cancer suppressing function whether compound is antagonist.With the same quadrat method of above-mentioned SCREENED COMPOUND, can filter out the acceptor disappearance thing and the analogue of antagonist action.
Polypeptide of the present invention can be directly used in disease treatment, for example, and various malignant tumours and cellular abnormality propagation etc.
Polypeptide of the present invention, and fragment, derivative, analogue or their cell can be used as antigen to produce antibody.These antibody can be polyclone or monoclonal antibody.Polyclonal antibody can obtain by the method with this polypeptide direct injection animal.The technology of preparation monoclonal antibody comprises hybridoma technology, three knurl technology, people B-quadroma technology, EBV-hybridoma technology etc.
Can be with polypeptide of the present invention and antagonist and suitable pharmaceutical carrier combination back use.These carriers can be water, glucose, ethanol, salt, damping fluid, glycerine and their combination.Composition comprises the polypeptide or the antagonist of safe and effective amount and carrier and the vehicle that does not influence effect of drugs.These compositions can be used as medicine and are used for disease treatment.
The present invention also provides medicine box or the test kit that contains one or more containers, and one or more medicinal compositions compositions of the present invention are housed in the container.With these containers, can have by the given indicative prompting of government authorities of making, using or selling medicine or biological products, the government authorities that this prompting reflects production, uses or sells permits it to use on human body.In addition, polypeptide of the present invention can be used in combination with other treatment compound.
Pharmaceutical composition can be with mode administration easily, as by in part, intravenously, intraperitoneal, intramuscular, subcutaneous, the nose or the route of administration of intracutaneous.Albumen with cancer suppressing function comes administration with the amount that treats and/or prevents concrete indication effectively.The proteic amount with cancer suppressing function and the dosage range that are applied to the patient will depend on many factors, as administering mode, person's to be treated healthiness condition and diagnostician's judgement.
The proteic polynucleotide of people with cancer suppressing function also can be used for multiple therapeutic purpose.Gene therapy technology can be used for treating since have that the proteic nothing of cancer suppressing function is expressed or the proteic expression with cancer suppressing function of unusual/non-activity due to cell proliferation, growth or metabolic disturbance.The gene therapy vector of reorganization can be used for treating the protein expression with cancer suppressing function or the disease of active caused by abnormal.Deriving from the expression vector of virus such as protein gene that retrovirus, adenovirus, adeno-associated virus (AAV), hsv, parvovirus etc. can be used for having cancer suppressing function is transferred in the cell.The method that structure carries the recombinant viral vector of the protein gene with cancer suppressing function be found in existing document (Sambrook, etal.).The people protein gene of reorganization with cancer suppressing function can be packaged in the liposome and be transferred in the cell in addition.
Suppress to have cancer suppressing function people's protein mRNA oligonucleotide (comprising sense-rna and DNA) and ribozyme also within the scope of the invention.Ribozyme is the enzyme sample RNA molecule that a kind of energy specificity is decomposed specific RNA, and its mechanism of action is to carry out the endonuclease effect after ribozyme molecule and the hybridization of complementary target RNA-specific.The RNA of antisense and DNA and ribozyme can obtain with existing any RNA or DNA synthetic technology, as the technology widespread use of solid phase phosphoamide chemical synthesis synthetic oligonucleotide.Antisense rna molecule can be transcribed acquisition by the dna sequence dna of this RNA that encodes in external or body.This dna sequence dna has been incorporated into the downstream of rna polymerase promoter of carrier.In order to increase the stability of nucleic acid molecule, available several different methods is modified it, and as increasing the sequence length of both sides, the connection between the ribonucleoside is used phosphoric acid thioester bond or peptide bond but not phosphodiester bond.
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.
Polypeptide of the present invention also can be used as the peptide spectrum analysis, for example, the polypeptide available physical, chemistry or enzyme carry out the specificity cutting, and carry out the two-dimentional or three-dimensional gel electrophoresis analysis of one dimension.
The present invention also provides the antibody at the people's proteantigen determinant with cancer suppressing function.These antibody include, but is not limited to: the fragment that polyclonal antibody, monoclonal antibody, chimeric antibody, single-chain antibody, Fab fragment and Fab expression library produce.These antibody can prepare with ordinary method.The anti-proteic antibody of people with cancer suppressing function can be used in the immunohistochemistry technology, detects the people's albumen with cancer suppressing function in the biopsy specimen.
With the also available labelled with radioisotope of the protein bound monoclonal antibody of the people with cancer suppressing function, inject in the body and can follow the tracks of its position and distribution.Antibody among the present invention can be used for treating or prevents and the relevant disease of people's albumen with cancer suppressing function.The antibody that gives suitable dosage can stimulate or block proteic generation of the people with cancer suppressing function or activity.
Antibody also can be used for designing the immunotoxin at a certain privileged sites in the body.As have cancer suppressing function people's albumen high-affinity monoclonal antibody can with bacterium or plant poison (as diphtheria toxin, ricin, abrine etc.) covalent attachment.
Available people's albumen or the polypeptide immune animal of the production of polyclonal antibody with cancer suppressing function, as rabbit, mouse, rat etc.Multiple adjuvant can be used for the enhancing immunity reaction, includes but not limited to freund's adjuvant etc.
Have cancer suppressing function people's protein monoclonal antibody can with hybridoma technology production (Kohler and Milstein.Nature, 1975,256:495-497).With the variable region bonded chimeric antibody in human constant region and inhuman source can with existing technology production (Morrison et al, PNAS, 1985,81:6851).And the technology of existing manufacture order chain antibody (U.S.PatNo.4946778) also can be used for producing the anti-proteic single-chain antibody of people with cancer suppressing function.
Can be incorporated into the rondom polypeptide storehouse that solid formation forms by the various amino acid that may make up by screening with the protein bound peptide molecule of the present invention obtains.During screening, must carry out mark to people's protein molecular with cancer suppressing function.
The invention still further relates to quantitatively and detection and localization has the diagnostic testing process of people's protein level of cancer suppressing function.These tests are known in the art, and comprise that FISH measures and radioimmunoassay.The people's protein level that is detected in the test with cancer suppressing function, the disease that can have the importance of people's albumen in various diseases of cancer suppressing function with laying down a definition and be used to diagnose albumen to work with cancer suppressing function.
Proteic polynucleotide with cancer suppressing function can be used for having the diagnosis and the treatment of the protein related diseases of cancer suppressing function.Aspect diagnosis, the proteic polynucleotide with cancer suppressing function can be used for detecting have cancer suppressing function proteic expression whether or under morbid state, have an abnormal exprssion of cancer suppressing function.As the protein D NA sequence with cancer suppressing function can be used for the hybridization of biopsy specimen is had with judgement the proteic abnormal expression of cancer suppressing function.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 the albumen with cancer suppressing function and also can detect proteic transcription product with cancer suppressing function.
The sudden change that detection has the protein gene of cancer suppressing function also can be used for diagnosing the relevant disease of albumen with cancer suppressing function.Form with protein mutation of cancer suppressing function comprises that to have point mutation that the protein D NA sequence of cancer suppressing function compares, transposition, disappearance, reorganization and other any unusual etc. with normal wild type.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.
Sequence of the present invention identifies it also is valuable to karyomit(e).These sequences can be specifically at certain bar human chromosome particular location and and can with its hybridization.At present, need to identify the concrete site of each gene on the karyomit(e).Yet have only chromosomal marker thing seldom to can be used for the marker chromosomes position now based on actual sequence data (repetition polymorphism).For these sequences are associated with disease related gene.The first step is positioned dna sequence dna of the present invention on the karyomit(e) exactly.
In brief, prepare PCR primer (preferred 15-35bp), sequence can be positioned on the karyomit(e) according to cDNA.Then, these primers are used for the somatocyte hybrid cell that the PCR screening contains each bar human chromosome.Have only those hybrid cells that contain corresponding to the people's gene of primer can produce the fragment of amplification.
The PCR localization method of somatocyte hybrid cell is that DNA is navigated to concrete chromosomal quick method.Use Oligonucleolide primers of the present invention,, can utilize one group to realize inferior location from specific chromosomal fragment or a large amount of genomic clone by similar approach.Other the similar strategy that can be used for chromosomal localization comprises in situ hybridization, uses the karyomit(e) prescreen and the hybridization preliminary election of the airflow classification of mark, thereby makes up the special cDNA storehouse of karyomit(e).
The cDNA clone is carried out fluorescence in situ hybridization (FISH) with Metaphase Chromosome, can in a step, accurately carry out chromosomal localization.The summary of this technology is referring to Verma etc., Human Chromosomes:a Manual of BasicTechniques, Pergamon Press, New York (1988).
In case sequence is positioned to chromosome position accurately, the physical location of this sequence on karyomit(e) just can be associated with the gene map data.These data for example are found in, V.Mckusick, Mendelian Inheritance in Man (can by with the online acquisition of Johns Hopkins University Welch Medical Library).Can pass through linkage analysis then, determine gene and navigated to relation between the disease on the chromosomal region already.
Then, need to measure ill and not cDNA between diseased individuals or genome sequence difference.If observe certain sudden change in some or all of diseased individuals, and this sudden change is not observed in any normal individual, then this sudden change may be the cause of disease of disease.More ill and diseased individuals not is usually directed at first seek the variation of structure in the karyomit(e), as from the horizontal visible of karyomit(e) or use based on detectable disappearance of the PCR of cDNA sequence or transposition.
Pyrenoids thuja acid full length sequence or its fragment with cancer suppressing function of the present invention can obtain with the method for pcr amplification method, recombination method or synthetic usually.For the pcr amplification method, can be disclosed according to the present invention about nucleotide sequence, 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.
At present, can be fully come the dna sequence dna of code book invention albumen (or its fragment, or derivatives thereof) by chemosynthesis.This dna sequence dna can be introduced then in the various dna moleculars (as carrier) and cell in this area.In addition, also can will suddenly change and introduce in the protein sequence of the present invention by chemosynthesis.
In addition, because the albumen with cancer suppressing function of the present invention has the natural acid sequence that is derived from the people, therefore, compare with the albumen of the same clan that derives from other species, estimate to have higher active and/or lower side effect (for example in the intravital immunogenicity of people lower or do not have) being applied to man-hour.
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:Cold Spring Harbor LaboratoryPress, 1989) condition described in, or the condition of advising according to manufacturer.
Embodiment
The acquisition of embodiment 1:cDNA gene and the restraining effect that the cancer cells clone is formed
FP17388, FP17814, FP18149, FP18376, FP18463 and FP18821 come from the human fetal cDNA library that makes up with ordinary method.Get fetal tissue, (GIBCO BRL company) extracts total RNA by manufacturer's specification sheets with Trizol reagent, extracts mRNA with the mRNA test kit (Pharmacia company) of purifying.Make up the cDNA library of above-mentioned mRNA with pCMV-script TMXRcDNA library construction test kit (Stratagene company).Wherein ThermoScript II is used MMLV-RT-Superscript II (GIBCO BRL) instead, and reverse transcription reaction carries out at 42 ℃.Transform XL 10-Gold recipient cell, obtained 1 * 10 6The cDNA library of cfu/ μ g cDNA titre.The first round is picking cDNA clone at random, is probe with high abundance cDNA clone with the cDNA clone who has proved cancer inhibitor cell growth function thereafter, screening by hybridization cDNA library, weak positive and negative clone of picking.With Qiagen 96 orifice plate plasmid extraction test kits, carry out the extraction of plasmid DNA by shop instruction.Plasmid DNA and empty carrier transfection simultaneously hepatoma cell line 7721.After the 100ng DNA alcohol precipitation drying, add 6 μ l H 2Transfection is treated in the O dissolving.Add 0.74 μ l liposome and 9.3 μ l serum-free mediums in every part of DNA sample, behind the mixing, room temperature was placed 10 minutes.Add 150 μ l serum-free mediums in every pipe, divide equally and add 3 holes and grow in 7721 cells of 96 orifice plates, placed 2 hours for 37 ℃, every hole adds 50 μ l serum-free mediums again, 37 ℃ 24 hours.Every hole is changed 100 μ l and is trained liquid entirely, 37 ℃ 24 hours, change the full training liquid 100 μ l that contain G418,37 ℃ 24-48 hour, the limit is observed, the training liquid that G418 concentration does not wait is changed on the limit.After about 2-3 time, there is the clone to form up to the microscopy cell, counting.Find that above-mentioned clone has the cell clone of inhibition formation effect, the result is as shown in the table.
CDNA clone's transfectional cell (7721) clone formation situation
Figure C0213640000121
The cDNA clone is adopted two deoxidation cessation method, on the ABI377DNA automatic sequencer, measure the nucleotide sequence of the nearly 500bp of one end.After the analysis, be defined as novel gene cloning, carry out the other end order-checking, do not obtain full length cDNA sequence yet, the design primer checks order once more, up to obtaining full length sequence (SEQ ID NO:1,4,7,10,13,16).
Embodiment 2: PCR obtains full-length gene from placenta or fetus cDNA
Get fetal tissue, (GIBCO BRL company) extracts total RNA by manufacturer's specification sheets with Trizol reagent, extracts mRNA with the mRNA test kit (Pharmacia company) of purifying.With MMLV-RT-Superscript II (GIBCO BRL), ThermoScript II is carried out reverse transcription reaction at 42 ℃, obtains placenta or fetus cDNA.Utilize the special primer (as shown in the table) of each gene, by 97 ℃ of 3 ' 1 circulations.94 ℃ 30 " 60 ℃ 30 " 72 ℃ of 1 ' 35 circulations, pcr amplification is carried out in 72 ℃ of 10 ' 1 circulations, and acquisition contains the amplified production of each protein gene of complete open reading frame sequence.Amplified production is through sequence verification, and the sequence that records with embodiment 1 conforms to, and changes amplified production over to host cell with routine techniques subsequently, obtains recombinant protein (SEQ ID NO:2,5,8,11,14,17).
Gene specific primer
Clone's title Special primer 1 (5 ' → 3 ') SEQ ID NO： Special primer 2 (3 ' → 5 ') SEQ ID NO：
FP17388 (82)ATTTCCTGACCTGTGATCTGCC 19 TCTTACCGAACTTGGGTCTCC(2107) 20
FP17814 (64)TTCTAGGTCCGTGCTGTTGTAA 21 CGTGTCCGTGAAATCGGGC(2731) 22
FP18149 (150)CTCCAAGACGTAGAGGAAGTGG 23 CCGAGCCGGAAGGATGGA(2240) 24
FP18376 (15)CTGTCAAATCTCAGGGTGCTG 25 TTGTCGTCCCTCCCGTTC(1366) 26
FP18463 (19)GGCACTGCTCACAGGACG 27 AGGCAGACAGGAAACAGGTG(1930) 28
FP18821 (136)TGAGTCAGCCTCGTAAATTGTG 29 TTGTTCTCGCTTTGAGGCAGA(2373) 30
Annotate: in the bracket is the correspondence position of primer in each gene DNA sequence.
Embodiment 3:cDNA cloned sequence is analyzed
1.FP17388
A: nucleotide sequence (SEQ ID NO:1) length: 2194 bases
B: aminoacid sequence (SEQ ID NO:3) length: 198 amino acid
C. Nucleotide and amino acid composite sequence (SEQ ID NO:2) clone number and protein name: FP17388 start code: 1019ATG stops coding: 1613TAG protein molecular weight: 22159.42
2.FP17814
A: nucleotide sequence (SEQ ID NO:4) length: 2834 bases
B: aminoacid sequence (SEQ ID NO:6) length: 208 amino acid
C. Nucleotide and amino acid composite sequence (SEQ ID NO:5) clone number and protein name: FP17814 start code: 1794ATG stops coding: 2418TGA protein molecular weight: 24154.66
3.FP18149
A: nucleotide sequence (SEQ ID NO:7) length: 2351 bases
B: aminoacid sequence (SEQ ID NO:9) length: 381 amino acid
C. Nucleotide and amino acid composite sequence (SEQ ID NO:8) clone number and protein name: FP18149 start code: 1061 ATG stop coding: 2204 TAG protein molecular weights: 41411.90
4.FP18376
A: nucleotide sequence (SEQ ID NO:10) length: 1413 bases
B: aminoacid sequence (SEQ ID NO:12) length: 117 amino acid
C. Nucleotide and amino acid composite sequence (SEQ ID NO:11) clone number and protein name: FP18376 start code: 58 ATG stop coding: 409 TAA protein molecular weights: 13159.52
5.FP18463
A: nucleotide sequence (SEQ ID NO:13) length: 2055 bases
B: aminoacid sequence (SEQ ID NO:15) length: 337 amino acid
C. Nucleotide and amino acid composite sequence (SEQ ID NO:14) clone number and protein name: FP18463 start code: 322 ATG stop coding: 1333 TAG protein molecular weights: 37576.65
6.FP18821
A: nucleotide sequence (SEQ ID NO:16) length: 2392 bases
B: aminoacid sequence (SEQ ID NO:18) length: 92 amino acid
C. Nucleotide and amino acid composite sequence (SEQ ID NO:17) clone number and protein name: FP18821 start code: 1089 ATG stop coding: 1365 TAA protein molecular weights: 9973.67
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 Xinshijie Gene Techn Development Co., Ltd.
＜120〉have new the people's albumen and the encoding sequence thereof of cancer suppressing function
<130>024038
<160>30
<170>PatentIn version 3.1
<210>1
<211>2194
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<400>1
gggcgcatgc cactacaccc ggctaatatt ttgtattttt agttgagaca ggattttacc 60
atgttggcca ggctggtctc gatttcctga cctgtgatct gcctgcctcg gcctcccaaa 120
gtgctgagat tacaggcgtg agccactgtg cccagccctt ggctactttt tatattttta 180
gtacagacag ggtttcatca tgtcggccag tctggtcttg aactcctgac cttgtgatac 240
actcacctcg gcctcgcaaa gtgctgggat tacaggcgtg agccaccgtg cctggccctt 300
ggctactttt tatattttta gtagagatgg ggtttcacca tgttggccag tctggtctcg 360
aactcctgac ctcaggtgat ccgccctcct cggcctccca aagcactggg attacaagcg 420
tgagccactg tgcctggccc aatcatagtt attttaaagc ccttgtttcc taactccaat 480
atgtggctta tctgtaatct gcttcttctg ttagctttcc gcatgattat tgatcactgt 540
ttcctgctgt gtcctgtatc tcgtgctttc tgtcagaggt atgcctcaaa ggaccgtggg 600
ggtccatatc tagggaccgt ggggggtcca tatgtcggga ccgtgggggt gtctatatct 660
cgggaccgtg ggggtccata tctaaggacc gtgggggtcc atatcttggg accatggggg 720
gtctatatgt caggaccgtg ggggtctata tctaaggacc gtggggttct gtatctcagg 780
accgtggggg tccatatcta gggaccgtgg gggtccatat ctagggaccg tggggggtcc 840
atatgtcggg accgtggggg tgtctatatc tcgggaccgt gggggtccat atctaaggac 900
tgtgggggtc catatcttgg agcccccaca agaaccacag gcccccttgg agcacccaca 960
ggagccacag ccccctttgc cacagctgga gcaggaacaa gctggcacac agcagcacat 1020
gggcttgcag cagcagacag gcacacagca gctggagcca catcccccac agccggaacc 1080
acagccaccc ttggatcccc cacaagagcc acagcccccc ttggagcccc cacaggagcc 1140
acaacccccc ttggatcccc cacaagagca cagcccccct tgcagcctcc acaggagcca 1200
cagcccccct tggagccccc agaagagcca cagccccctt tgccacagct ggagcaggaa 1260
caggttggca cacggcagca cacgggcttg cagcagcaga cgggcacaca gcagctggag 1320
ccagaacctc cacagccaga gccacagccc ccacagccgg agccacagcc cccacagccg 1380
gagccacagc ccccacagct ggagccacag cctccggagc agccgcaaca gcccatggtt 1440
ctggtggatt gagggtggag caggtagagg agcaggtgag agggaggtgc aggtgtggag 1500
ctccctgagc ctggaccctt tatatccctg cccagggtca tgtgtgaggc tgggcacaca 1560
tttcctggtt cctgtttgtg ccatttttag ggcccctttt tcttgtttcc tctagaaatc 1620
cgccccttgg tgtatgggct gctcagtggg ctgctgctct cttgctgaat ctgtgtccag 1680
acttaatgga ggcccccaag ggtctagcct ctccctgttg actccagagt cacactggat 1740
ttacaaaagc atctatttta ggctgggcat ggtggctcac acctgtaatc tcagcacttt 1800
gggaggccta ggcaggtgga tcacttgagg tgaggggttg gagaccagcc tggccaaaat 1860
ggtgaaatct cgtgtctact aaaaacacaa aaattaactg ggtgtggtgg ctcacatctg 1920
taatcccagc actttgggag gccgaggcag gtggatcatt tgaggtcagg agttggagac 1980
cagcctgccc aacatggcaa aaccccatct ttactaaaaa tacaaaaatt agccaggcat 2040
ggtggtgcat gcttgtaact ccagctactt gggaggctga ggcaggagaa tggcttgaac 2100
ccagaggtga aggttgcagt gagcagagat cacaccactg cactctagtg tgggcaacag 2160
agcggggctc tgtctcaaaa aaaaaaaaaa aaaa 2194
<210>2
<211>2194
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<220>
<221>CDS
<222>(1019)..(1612)
<223>
<400>2
gggcgcatgc cactacaccc ggctaatatt ttgtattttt agttgagaca ggattttacc 60
atgttggcca ggctggtctc gatttcctga cctgtgatct gcctgcctcg gcctcccaaa 120
gtgctgagat tacaggcgtg agccactgtg cccagccctt ggctactttt tatattttta 180
gtacagacag ggtttcatca tgtcggccag tctggtcttg aactcctgac cttgtgatac 240
actcacctcg gcctcgcaaa gtgctgggat tacaggcgtg agccaccgtg cctggccctt 300
ggctactttt tatattttta gtagagatgg ggtttcacca tgttggccag tctggtctcg 360
aactcctgac ctcaggtgat ccgccctcct cggcctccca aagcactggg attacaagcg 420
tgagccactg tgcctggccc aatcatagtt attttaaagc ccttgtttcc taactccaat 480
atgtggctta tctgtaatct gcttcttctg ttagctttcc gcatgattat tgatcactgt 540
ttcctgctgt gtcctgtatc tcgtgctttc tgtcagaggt atgcctcaaa ggaccgtggg 600
ggtccatatc tagggaccgt ggggggtcca tatgtcggga ccgtgggggt gtctatatct 660
cgggaccgtg ggggtccata tctaaggacc gtgggggtcc atatcttggg accatggggg 720
gtctatatgt caggaccgtg ggggtctata tctaaggacc gtggggttct gtatctcagg 780
accgtggggg tccatatcta gggaccgtgg gggtccatat ctagggaccg tggggggtcc 840
atatgtcggg accgtggggg tgtctatatc tcgggaccgt gggggtccat atctaaggac 900
tgtgggggtc catatcttgg agcccccaca agaaccacag gcccccttgg agcacccaca 960
ggagccacag ccccctttgc cacagctgga gcaggaacaa gctggcacac agcagcac 1018
atg ggc ttg cag cag cag aca ggc aca cag cag ctg gag cca cat ccc 1066
Met Gly Leu Gln Gln Gln Thr Gly Thr Gln Gln Leu Glu Pro His Pro
1 5 10 15
cca cag ccg gaa cca cag cca ccc ttg gat ccc cca caa gag cca cag 1114
Pro Gln Pro Glu Pro Gln Pro Pro Leu Asp Pro Pro Gln Glu Pro Gln
20 25 30
ccc ccc ttg gag ccc cca cag gag cca caa ccc ccc ttg gat ccc cca 1162
Pro Pro Leu Glu Pro Pro Gln Glu Pro Gln Pro Pro Leu Asp Pro Pro
35 40 45
caa gag cac agc ccc cct tgc agc ctc cac agg agc cac agc ccc cct 1210
Gln Glu His Ser Pro Pro Cys Ser Leu His Arg Ser His Ser Pro Pro
50 55 60
tgg agc ccc cag aag agc cac agc ccc ctt tgc cac agc tgg agc agg 1258
Trp Ser Pro Gln Lys Ser His Ser Pro Leu Cys His Ser Trp Ser Arg
65 70 75 80
aac agg ttg gca cac ggc agc aca cgg gct tgc agc agc aga cgg gca 1306
Asn Arg Leu Ala His Gly Ser Thr Arg Ala Cys Ser Ser Arg Arg Ala
85 90 95
cac agc agc tgg agc cag aac ctc cac agc cag agc cac agc ccc cac 1354
His Ser Ser Trp Ser Gln Asn Leu His Ser Gln Ser His Ser Pro His
100 105 110
agc cgg agc cac agc ccc cac agc cgg agc cac agc ccc cac agc tgg 1402
Ser Arg Ser His Ser Pro His Ser Arg Ser His Ser Pro His Ser Trp
115 120 125
agc cac agc ctc cgg agc agc cgc aac agc cca tgg ttc tgg tgg att 1450
Ser His Ser Leu Arg Ser Ser Arg Asn Ser Pro Trp Phe Trp Trp Ile
130 135 140
gag ggt gga gca ggt aga gga gca ggt gag agg gag gtg cag gtg tgg 1498
Glu Gly Gly Ala Gly Arg Gly Ala Gly Glu Arg Glu Val Gln Val Trp
145 150 155 160
agc tcc ctg agc ctg gac cct tta tat ccc tgc cca ggg tca tgt gtg 1546
Ser Ser Leu Ser Leu Asp Pro Leu Tyr Pro Cys Pro Gly Ser Cys Val
165 170 175
agg ctg ggc aca cat ttc ctg gtt cct gtt tgt gcc att ttt agg gcc 1594
Arg Leu Gly Thr His Phe Leu Val Pro Val Cys Ala Ile Phe Arg Ala
180 185 190
cct ttt tct tgt ttc ctc tagaaatccg ccccttggtg tatgggctgc 1642
Pro Phe Ser Cys Phe Leu
195
tcagtgggct gctgctctct tgctgaatct gtgtccagac ttaatggagg cccccaaggg 1702
tctagcctct ccctgttgac tccagagtca cactggattt acaaaagcat ctattttagg 1762
ctgggcatgg tggctcacac ctgtaatctc agcactttgg gaggcctagg caggtggatc 1822
acttgaggtg aggggttgga gaccagcctg gccaaaatgg tgaaatctcg tgtctactaa 1882
aaacacaaaa attaactggg tgtggtggct cacatctgta atcccagcac tttgggaggc 1942
cgaggcaggt ggatcatttg aggtcaggag ttggagacca gcctgcccaa catggcaaaa 2002
ccccatcttt actaaaaata caaaaattag ccaggcatgg tggtgcatgc ttgtaactcc 2062
agctacttgg gaggctgagg caggagaatg gcttgaaccc agaggtgaag gttgcagtga 2122
gcagagatca caccactgca ctctagtgtg ggcaacagag cggggctctg tctcaaaaaa 2182
aaaaaaaaaa aa 2194
<210>3
<211>198
<212>PRT
＜213〉homo sapiens (Homo sapiens)
<400>3
Met Gly Leu Gln Gln Gln Thr Gly Thr Gln Gln Leu Glu Pro His Pro
1 5 10 15
Pro Gln Pro Glu Pro Gln Pro Pro Leu Asp Pro Pro Gln Glu Pro Gln
20 25 30
Pro Pro Leu Glu Pro Pro Gln Glu Pro Gln Pro Pro Leu Asp Pro Pro
35 40 45
Gln Glu His Ser Pro Pro Cys Ser Leu His Arg Ser His Ser Pro Pro
50 55 60
Trp Ser Pro Gln Lys Ser His Ser Pro Leu Cys His Ser Trp Ser Arg
65 70 75 80
Asn Arg Leu Ala His Gly Ser Thr Arg Ala Cys Ser Ser Arg Arg Ala
85 90 95
His Ser Ser Trp Ser Gln Asn Leu His Ser Gln Ser His Ser Pro His
100 105 110
Ser Arg Ser His Ser Pro His Ser Arg Ser His Ser Pro His Ser Trp
115 120 125
Ser His Ser Leu Arg Ser Ser Arg Asn Ser Pro Trp Phe Trp Trp Ile
130 135 140
Glu Gly Gly Ala Gly Arg Gly Ala Gly Glu Arg Glu Val Gln Val Trp
145 150 155 160
Ser Ser Leu Ser Leu Asp Pro Leu Tyr Pro Cys Pro Gly Ser Cys Val
165 170 175
Arg Leu Gly Thr His Phe Leu Val Pro Val Cys Ala Ile Phe Arg Ala
180 185 190
Pro Phe Ser Cys Phe Leu
195
<210>4
<211>2834
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<400>4
gcccaaatcc tcttctccaa gaaggtaggc tccacccgct ttgcccttct ccactcgctt 60
tttttctagg tccgtgctgt tgtaagtcat ttgagggacc tgcgtacaat cccagaccta 120
ctcgtggggc atgtgtgacc ctgtttaggc ttgggtactc tgcctctcct tttgctcctc 180
tctgtgaaat gggcagagga tgcccattca aggaaaaaga gcagggagca taagacgcac 240
ggtcgtgggg acggctgacc cctctctccc tttgcttctt gggagatccc tgccagggtg 300
caagatgtgg attcgagttg cttgaaggga aatggcattt cctctggtgc cagaggacag 360
ggcctgacag cctactcacc cacttcgtca ctgctcccgg ctttcccttc gttatgtctc 420
ctgggacttc ccatggctgg accccgtcct gggcaattct gagacccaga gataagtcag 480
atccagaccc acctaagccc tgacccttac tcactttttg agaaactgat agatgaggtc 540
gaaaacgcta ggaagtgttg gaactttttg cacccagaca gtctgattcc agatccccaa 600
cgcttcctgg gagagcagac atttgagctg ggactcccag gatatgaaag gacaagaagg 660
tatttcaggg actcaaggca cacggagaag ttggatgcct ctgtaggggg gttctttagt 720
ggggatgaga cctccattgg agctgatgag ttccacccag gggccggaag tcagtcggca 780
cccacagagc tagtgacgag acactggaag gctgagctga ccacctgcgc tgccccaggg 840
caggaggatg aagatgaaga tgaagatgac tctccacctt gctccacagc caaacacagc 900
caaggcacca ggaagttgcg ttacatcact tccgctttcc aggactccag tgggggtgac 960
tcatgtgctt ctgaggagga accaaccttt gaccccggct atgaacctga ctgggctgtc 1020
atcagcactg tgcggccaca gctctgccac tcagagccca cgagaggtgc agagcccagc 1080
tgcccccagg gctcctgggc cttctcagtg agtctggggg agctaaagtc catccgccgc 1140
tccaagccag gcctcagctg ggcctacctg gttctggtga cccaggctgg aggttccctg 1200
cccgcactgc acttccaccg cgggggcacc cgcgccctgc tccgcgtcct cagccgctac 1260
ctgctgttgg ccagctcccc gcaggactcc cgcctctacc ttgtcttccc ccacgactcc 1320
tctgctctct ccaactcctt ccaccacctg cagctctttg accaggacag ctccatgtgg 1380
tgtcacgctt cctccaggat ccctactcca ccaccttcag cagcttctcc cgagtgacca 1440
acttcttccg gggtgccctg cagccacagc ctgagggagc cgcctccgac cttcccccgc 1500
cacccgacga tgagcccgag cctggattcg aggtcatttc ctgtgtggag ctggggcctc 1560
ggccaaccgt ggagcggggc cctccagtta cagaggagga gtgggcacgc cacgtgggcc 1620
ctgaaggtcg cctgcagcag gtccctgagc tgaagaaccg gatcttctcg gggggtctga 1680
gccccagcct gcggcgcgag gcctggaagt tcctcctagg gtacctcagc tgggaaggca 1740
cagctgagga gcacaaggcc cacatacgca agaaaacgga tgagtatttc cgcatgaagc 1800
tgcagtggaa atctgtgagc cctgagcagg agcggagaaa ctcacttctg catggatacc 1860
gcagcctcat cgaaagggat gtgagccgca ctgacaggac caacaagttc tacgagggtc 1920
ccgagaaccc ggggctgggc ctgctgaacg atatcctcct cacctactgc atgtatcact 1980
tcgacctcgg ctacgtccag ggcatgagtg atcttctctc cccgatcctc tacgtcattc 2040
agaacgaggt ggatgctttc tggtgtttct gtggcttcat ggagctcgtg caagggaact 2100
ttgaagagag ccaggagacc atgaagcggc aactcgggcg actgctgctg ctcctgaggg 2160
tgctggaccc cctgctctgc gacttcctgg attcccagga ctccggctct ctctgcttct 2220
gtttccggtg gctgctcatc tggttcaaga gggaattccc cttcccggat gtccttcggc 2280
tgtgggaggt gctgtggaca gggctccctg gccccaatct gcacctgctg gtggcctgcg 2340
ccatcctgga catggagagg gacaccctca tgctgtccgg cttcggctcc atgagatcct 2400
caagcacatc aacgagctga ctatgaagct gagcgtggag gacgtgctga cccgcgccga 2460
ggccctgcac cgccagctaa ccgcctgccc cgagctgccc cacaacgtgc aggagatcct 2520
ggggctggcc ccgcccgcag agccccacag ccctcgccac cgcctccccg ctgcctctgt 2580
cgcccacccg ggccccgccc accccgccgc cctccacgga cacagccccg cagcccgaca 2640
gcagcctgga gatccttccc gaggaggagg acgagggcgc cgactcctaa ccccgccagg 2700
cagcctcgtt ctgcacaggc actttagccc gagccaggca cacctgcgag ggggcaggtg 2760
tgctcccccg ccctgctgat aagctggctt cattaaactg acacttctca tgtgaaaaaa 2820
aaaaaaaaaa aaaa 2834
<210>5
<211>2834
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<220>
<221>CDS
<222>(1794)..(2417)
<223>
<400>5
gcccaaatcc tcttctccaa gaaggtaggc tccacccgct ttgcccttct ccactcgctt 60
tttttctagg tccgtgctgt tgtaagtcat ttgagggacc tgcgtacaat cccagaccta 120
ctcgtggggc atgtgtgacc ctgtttaggc ttgggtactc tgcctctcct tttgctcctc 180
tctgtgaaat gggcagagga tgcccattca aggaaaaaga gcagggagca taagacgcac 240
ggtcgtgggg acggctgacc cctctctccc tttgcttctt gggagatccc tgccagggtg 300
caagatgtgg attcgagttg cttgaaggga aatggcattt cctctggtgc cagaggacag 360
ggcctgacag cctactcacc cacttcgtca ctgctcccgg ctttcccttc gttatgtctc 420
ctgggacttc ccatggctgg accccgtcct gggcaattct gagacccaga gataagtcag 480
atccagaccc acctaagccc tgacccttac tcactttttg agaaactgat agatgaggtc 540
gaaaacgcta ggaagtgttg gaactttttg cacccagaca gtctgattcc agatccccaa 600
cgcttcctgg gagagcagac atttgagctg ggactcccag gatatgaaag gacaagaagg 660
tatttcaggg actcaaggca cacggagaag ttggatgcct ctgtaggggg gttctttagt 720
ggggatgaga cctccattgg agctgatgag ttccacccag gggccggaag tcagtcggca 780
cccacagagc tagtgacgag acactggaag gctgagctga ccacctgcgc tgccccaggg 840
caggaggatg aagatgaaga tgaagatgac tctccacctt gctccacagc caaacacagc 900
caaggcacca ggaagttgcg ttacatcact tccgctttcc aggactccag tgggggtgac 960
tcatgtgctt ctgaggagga accaaccttt gaccccggct atgaacctga ctgggctgtc 1020
atcagcactg tgcggccaca gctctgccac tcagagccca cgagaggtgc agagcccagc 1080
tgcccccagg gctcctgggc cttctcagtg agtctggggg agctaaagtc catccgccgc 1140
tccaagccag gcctcagctg ggcctacctg gttctggtga cccaggctgg aggttccctg 1200
cccgcactgc acttccaccg cgggggcacc cgcgccctgc tccgcgtcct cagccgctac 1260
ctgctgttgg ccagctcccc gcaggactcc cgcctctacc ttgtcttccc ccacgactcc 1320
tctgctctct ccaactcctt ccaccacctg cagctctttg accaggacag ctccatgtgg 1380
tgtcacgctt cctccaggat ccctactcca ccaccttcag cagcttctcc cgagtgacca 1440
acttcttccg gggtgccctg cagccacagc ctgagggagc cgcctccgac cttcccccgc 1500
cacccgacga tgagcccgag cctggattcg aggtcatttc ctgtgtggag ctggggcctc 1560
ggccaaccgt ggagcggggc cctccagtta cagaggagga gtgggcacgc cacgtgggcc 1620
ctgaaggtcg cctgcagcag gtccctgagc tgaagaaccg gatcttctcg gggggtctga 1680
gccccagcct gcggcgcgag gcctggaagt tcctcctagg gtacctcagc tgggaaggca 1740
cagctgagga gcacaaggcc cacatacgca agaaaacgga tgagtatttc cgc atg 1796
Met
1
aag ctg cag tgg aaa tct gtg agc cct gag cag gag cgg aga aac tca 1844
Lys Leu Gln Trp Lys Ser Val Ser Pro Glu Gln Glu Arg Arg Asn Ser
5 10 15
ctt ctg cat gga tac cgc agc ctc atc gaa agg gat gtg agc cgc act 1892
Leu Leu His Gly Tyr Arg Ser Leu Ile Glu Arg Asp Val Ser Arg Thr
20 25 30
gac agg acc aac aag ttc tac gag ggt ccc gag aac ccg ggg ctg ggc 1940
Asp Arg Thr Asn Lys Phe Tyr Glu Gly Pro Glu Asn Pro Gly Leu Gly
35 40 45
ctg ctg aac gat atc ctc ctc acc tac tgc atg tat cac ttc gac ctc 1988
Leu Leu Asn Asp Ile Leu Leu Thr Tyr Cys Met Tyr His Phe Asp Leu
50 55 60 65
ggc tac gtc cag ggc atg agt gat ctt ctc tcc ccg atc ctc tac gtc 2036
Gly Tyr Val Gln Gly Met Ser Asp Leu Leu Ser Pro Ile Leu Tyr Val
70 75 80
att cag aac gag gtg gat gct ttc tgg tgt ttc tgt ggc ttc atg gag 2084
Ile Gln Asn Glu Val Asp Ala Phe Trp Cys Phe Cys Gly Phe Met Glu
85 90 95
ctc gtg caa ggg aac ttt gaa gag agc cag gag acc atg aag cgg caa 2132
Leu Val Gln Gly Asn Phe Glu Glu Ser Gln Glu Thr Met Lys Arg Gln
100 105 110
ctc ggg cga ctg ctg ctg ctc ctg agg gtg ctg gac ccc ctg ctc tgc 2180
Leu Gly Arg Leu Leu Leu Leu Leu Arg Val Leu Asp Pro Leu Leu Cys
115 120 125
gac ttc ctg gat tcc cag gac tcc ggc tct ctc tgc ttc tgt ttc cgg 2228
Asp Phe Leu Asp Ser Gln Asp Ser Gly Ser Leu Cys Phe Cys Phe Arg
130 135 140 145
tgg ctg ctc atc tgg ttc aag agg gaa ttc ccc ttc ccg gat gtc ctt 2276
Trp Leu Leu Ile Trp Phe Lys Arg Glu Phe Pro Phe Pro Asp Val Leu
150 155 160
cgg ctg tgg gag gtg ctg tgg aca ggg ctc cct ggc ccc aat ctg cac 2324
Arg Leu Trp Glu Val Leu Trp Thr Gly Leu Pro Gly Pro Asn Leu His
165 170 175
ctg ctg gtg gcc tgc gcc atc ctg gac atg gag agg gac acc ctc atg 2372
Leu Leu Val Ala Cys Ala Ile Leu Asp Met Glu Arg Asp Thr Leu Met
180 185 190
ctg tcc ggc ttc ggc tcc atg aga tcc tca agc aca tca acg agc 2417
Leu Ser Gly Phe Gly Ser Met Arg Ser Ser Ser Thr Ser Thr Ser
195 200 205
tgactatgaa gctgagcgtg gaggacgtgc tgacccgcgc cgaggccctg caccgccagc 2477
taaccgcctg ccccgagctg ccccacaacg tgcaggagat cctggggctg gccccgcccg 2537
cagagcccca cagccctcgc caccgcctcc ccgctgcctc tgtcgcccac ccgggccccg 2597
cccaccccgc cgccctccac ggacacagcc ccgcagcccg acagcagcct ggagatcctt 2657
cccgaggagg aggacgaggg cgccgactcc taaccccgcc aggcagcctc gttctgcaca 2717
ggcactttag cccgagccag gcacacctgc gagggggcag gtgtgctccc ccgccctgct 2777
gataagctgg cttcattaaa ctgacacttc tcatgtgaaa aaaaaaaaaa aaaaaaa 2834
<210>6
<211>208
<212>PRT
＜213〉homo sapiens (Homo sapiens)
<400>6
Met Lys Leu Gln Trp Lys Ser Val Ser Pro Glu Gln Glu Arg Arg Asn
1 5 10 15
Ser Leu Leu His Gly Tyr Arg Ser Leu Ile Glu Arg Asp Val Ser Arg
20 25 30
Thr Asp Arg Thr Asn Lys Phe Tyr Glu Gly Pro Glu Asn Pro Gly Leu
35 40 45
Gly Leu Leu Asn Asp Ile Leu Leu Thr Tyr Cys Met Tyr His Phe Asp
50 55 60
Leu Gly Tyr Val Gln Gly Met Ser Asp Leu Leu Ser Pro Ile Leu Tyr
65 70 75 80
Val Ile Gln Asn Glu Val Asp Ala Phe Trp Cys Phe Cys Gly Phe Met
85 90 95
Glu Leu Val Gln Gly Asn Phe Glu Glu Ser Gln Glu Thr Met Lys Arg
100 105 110
Gln Leu Gly Arg Leu Leu Leu Leu Leu Arg Val Leu Asp Pro Leu Leu
115 120 125
Cys Asp Phe Leu Asp Ser Gln Asp Ser Gly Ser Leu Cys Phe Cys Phe
130 135 140
Arg Trp Leu Leu Ile Trp Phe Lys Arg Glu Phe Pro Phe Pro Asp Val
145 150 155 160
Leu Arg Leu Trp Glu Val Leu Trp Thr Gly Leu Pro Gly Pro Asn Leu
165 170 175
His Leu Leu Val Ala Cys Ala Ile Leu Asp Met Glu Arg Asp Thr Leu
180 185 190
Met Leu Ser Gly Phe Gly Ser Met Arg Ser Ser Ser Thr Ser Thr Ser
195 200 205
<210>7
<211>2351
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<400>7
ggtgtcctgt gtcagggcca cagcccggga gcacgtggcg gtcagagagg agcggaggca 60
gtggtccaga gggtggaggt gagccccagc ctccctgacc cccaggctcc cctggctcca 120
gccctaaggt gctctcccag ccgtccgacc tggatctcca agacgtagag gaagtggaga 180
tcggcagaga caccttctgg cccggtgagt gagcagaatc gggttactgg gggaccagag 240
ctctgcgcgt gccaggctcc tcacaatgcc cacctctgtg gcacccccaa tgctggcacc 300
acccccttcc tggggcccct tttcttccct ggcatcagct cacctctgcc ccgtggcccc 360
tcccccactc taatgccagt ccagttactg atgcttcccg aggcggtgac tgcagcccac 420
agccacctgc gccagtggcc ccacctggac tcctgccggc cccagctgga cacagtgaat 480
caggaactcc tccctcccct ccacctgcct tgggctgggg cttctggctc agggcaccac 540
tgtcccccca gatccacgag ttatcctgtg tcccctcctc ctcctgcatc ccctagatgg 600
cctcccctca gcctggttgg agctgtgtcc aggtcccctg cacctccccc ttgagaagct 660
cccagggttc cctctgggcg atgttagtga tatggctgga tttaacagtg gtgagccctc 720
gcccacatct gggcggccct ggggtgggga gcaggtgtct gaggtcaggt ccctggaaag 780
agaccctggg ccagagattt gcgtgggaag ctcctggggt gggcatggga gctgggcggc 840
agcacacttg ccaccgaggc ctggggcagc cccatgagga gctgtgccca tatggaccac 900
atcctcggca agggctttga ggcgccctct ggagctctgg cgtcccccag gttccgggtc 960
ctcaccaaca cttagtgctt cctcctggct aagggtgacc ccacggtctg tcttgccact 1020
gtggctccgt gcaccggcac ggccgccgtg gtgactgtgg atggtcccag cagctctggg 1080
tcaccctgca acgcccccca cccctgtgtc tatgtcagct ctcctgggca cggcactccg 1140
agcccctgtc cgcttgtctc ccatccactg ggtgcctggg ccctccttgt cccctctaag 1200
cccccctgtc cagctctgct ctcgctcccc gtccccgtcc ctccgtgcca cggtgggggc 1260
atgcgggtac tatccgcctc tgccgtttct catttcagac tccgagccca agccggagca 1320
ggctccacgc tctcctggct ctcaggcccc tgacgagggg gcgggcgggg cgctgcgcag 1380
cctcctgagg agccttcccc gcagggcccg gtgcagcgcc ggcttcgggc ctgaatccag 1440
cgcggagcgg ccggcgggcc agccgcctgg ggccgtccct tgcgcccagc cgcggggcgc 1500
ctggcgcgtg acgctcgtgc agcaagcagc ggccgggccc gagggtgcgc ccgagcgggc 1560
tgccgagctg ggagtcaact tcggtcggag ccggcagggc agcgcgcggg ggaccaagcc 1620
gcacaggtgc gaggcctgcg gcaagagttt caagtataac tcgctgctcc tgaagcacca 1680
gcgcatccac acgggcgaga agccctacgc ctgccacgag tgcggcaagc gcttccgcgg 1740
ctggtcgggc ttcatccagc accaccgcat ccacacgggc gagaagccct acgagtgcgg 1800
ccagtgcggc cgcgccttca gccacagctc gcacttcacg cagcacctgc gcatccacaa 1860
cggcgagaag ccctacaagt gcggcgagtg cggccaggcc ttcagccaga gctccaacct 1920
ggtgcgccac cagcggctgc acacgggtga gaagccctac gcctgcagcc agtgcggcaa 1980
ggccttcatc tggagctccg tgctcatcga gcaccagcgc atccacactg gcgagaagcc 2040
ctacgagtgc tccgactgcg gcaaagcctt ccgcggccgc tcgcacttct tccggcacct 2100
gcggacccac acgggcgaga agcccttcgc gtgtggcgcc tgcggcaagg ccttcggcca 2160
gagctcccag ctcatccagc accagcgggt gcactaccgc gagtagccgg gcgggggctc 2220
ggggctcggc cttcctacct gcccccaacc caccctccac cccgtccccc acggtgggca 2280
ctgcccagca ccgcatgcca cgtgtccgga ataaattctt tttgattgtt ggaaaaaaaa 2340
aaaaaaaaaa a 2351
<210>8
<211>2351
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<220>
<221>CDS
<222>(1061)..(2203)
<223>
<400>8
ggtgtcctgt gtcagggcca cagcccggga gcacgtggcg gtcagagagg agcggaggca 60
gtggtccaga gggtggaggt gagccccagc ctccctgacc cccaggctcc cctggctcca 120
gccctaaggt gctctcccag ccgtccgacc tggatctcca agacgtagag gaagtggaga 180
tcggcagaga caccttctgg cccggtgagt gagcagaatc gggttactgg gggaccagag 240
ctctgcgcgt gccaggctcc tcacaatgcc cacctctgtg gcacccccaa tgctggcacc 300
acccccttcc tggggcccct tttcttccct ggcatcagct cacctctgcc ccgtggcccc 360
tcccccactc taatgccagt ccagttactg atgcttcccg aggcggtgac tgcagcccac 420
agccacctgc gccagtggcc ccacctggac tcctgccggc cccagctgga cacagtgaat 480
caggaactcc tccctcccct ccacctgcct tgggctgggg cttctggctc agggcaccac 540
tgtcccccca gatccacgag ttatcctgtg tcccctcctc ctcctgcatc ccctagatgg 600
cctcccctca gcctggttgg agctgtgtcc aggtcccctg cacctccccc ttgagaagct 660
cccagggttc cctctgggcg atgttagtga tatggctgga tttaacagtg gtgagccctc 720
gcccacatct gggcggccct ggggtgggga gcaggtgtct gaggtcaggt ccctggaaag 780
agaccctggg ccagagattt gcgtgggaag ctcctggggt gggcatggga gctgggcggc 840
agcacacttg ccaccgaggc ctggggcagc cccatgagga gctgtgccca tatggaccac 900
atcctcggca agggctttga ggcgccctct ggagctctgg cgtcccccag gttccgggtc 960
ctcaccaaca cttagtgctt cctcctggct aagggtgacc ccacggtctg tcttgccact 1020
gtggctccgt gcaccggcac ggccgccgtg gtgactgtgg atg gtc cca gca gct 1075
Met Val Pro Ala Ala
1 5
ctg ggt cac cct gca acg ccc ccc acc cct gtg tct atg tca gct ctc 1123
Leu Gly His Pro Ala Thr Pro Pro Thr Pro Val Ser Met Ser Ala Leu
10 15 20
ctg ggc acg gca ctc cga gcc cct gtc cgc ttg tct ccc atc cac tgg 1171
Leu Gly Thr Ala Leu Arg Ala Pro Val Arg Leu Ser Pro Ile His Trp
25 30 35
gtg cct ggg ccc tcc ttg tcc cct cta agc ccc cct gtc cag ctc tgc 1219
Val Pro Gly Pro Ser Leu Ser Pro Leu Ser Pro Pro Val Gln Leu Cys
40 45 50
tct cgc tcc ccg tcc ccg tcc ctc cgt gcc acg gtg ggg gca tgc ggg 1267
Ser Arg Ser Pro Ser Pro Ser Leu Arg Ala Thr Val Gly Ala Cys Gly
55 60 65
tac tat ccg cct ctg ccg ttt ctc att tca gac tcc gag ccc aag ccg 1315
Tyr Tyr Pro Pro Leu Pro Phe Leu Ile Ser Asp Ser Glu Pro Lys Pro
70 75 80 85
gag cag gct cca cgc tct cct ggc tct cag gcc cct gac gag ggg gcg 1363
Glu Gln Ala Pro Arg Ser Pro Gly Ser Gln Ala Pro Asp Glu Gly Ala
90 95 100
ggc ggg gcg ctg cgc agc ctc ctg agg agc ctt ccc cgc agg gcc cgg 1411
Gly Gly Ala Leu Arg Ser Leu Leu Arg Ser Leu Pro Arg Arg Ala Arg
105 110 115
tgc agc gcc ggc ttc ggg cct gaa tcc agc gcg gag cgg ccg gcg ggc 1459
Cys Ser Ala Gly Phe Gly Pro Glu Ser Ser Ala Glu Arg Pro Ala Gly
120 125 130
cag ccg cct ggg gcc gtc cct tgc gcc cag ccg cgg ggc gcc tgg cgc 1507
Gln Pro Pro Gly Ala Val Pro Cys Ala Gln Pro Arg Gly Ala Trp Arg
135 140 145
gtg acg ctc gtg cag caa gca gcg gcc ggg ccc gag ggt gcg ccc gag 1555
Val Thr Leu Val Gln Gln Ala Ala Ala Gly Pro Glu Gly Ala Pro Glu
150 155 160 165
cgg gct gcc gag ctg gga gtc aac ttc ggt cgg agc cgg cag ggc agc 1603
Arg Ala Ala Glu Leu Gly Val Asn Phe Gly Arg Ser Arg Gln Gly Ser
170 175 180
gcg cgg ggg acc aag ccg cac agg tgc gag gcc tgc ggc aag agt ttc 1651
Ala Arg Gly Thr Lys Pro His Arg Cys Glu Ala Cys Gly Lys Ser Phe
185 190 195
aag tat aac tcg ctg ctc ctg aag cac cag cgc atc cac acg ggc gag 1699
Lys Tyr Asn Ser Leu Leu Leu Lys His Gln Arg Ile His Thr Gly Glu
200 205 210
aag ccc tac gcc tgc cac gag tgc ggc aag cgc ttc cgc ggc tgg tcg 1747
Lys Pro Tyr Ala Cys His Glu Cys Gly Lys Arg Phe Arg Gly Trp Ser
215 220 225
ggc ttc atc cag cac cac cgc atc cac acg ggc gag aag ccc tac gag 1795
Gly Phe Ile Gln His His Arg Ile His Thr Gly Glu Lys Pro Tyr Glu
230 235 240 245
tgc ggc cag tgc ggc cgc gcc ttc agc cac agc tcg cac ttc acg cag 1843
Cys Gly Gln Cys Gly Arg Ala Phe Ser His Ser Ser His Phe Thr Gln
250 255 260
cac ctg cgc atc cac aac ggc gag aag ccc tac aag tgc ggc gag tgc 1891
His Leu Arg Ile His Asn Gly Glu Lys Pro Tyr Lys Cys Gly Glu Cys
265 270 275
ggc cag gcc ttc agc cag agc tcc aac ctg gtg cgc cac cag cgg ctg 1939
Gly Gln Ala Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Leu
280 285 290
cac acg ggt gag aag ccc tac gcc tgc agc cag tgc ggc aag gcc ttc 1987
His Thr Gly Glu Lys Pro Tyr Ala Cys Ser Gln Cys Gly Lys Ala Phe
295 300 305
atc tgg agc tcc gtg ctc atc gag cac cag cgc atc cac act ggc gag 2035
Ile Trp Ser Ser Val Leu Ile Glu His Gln Arg Ile His Thr Gly Glu
310 315 320 325
aag ccc tac gag tgc tcc gac tgc ggc aaa gcc ttc cgc ggc cgc tcg 2083
Lys Pro Tyr Glu Cys Ser Asp Cys Gly Lys Ala Phe Arg Gly Arg Ser
330 335 340
cac ttc ttc cgg cac ctg cgg acc cac acg ggc gag aag ccc ttc gcg 2131
His Phe Phe Arg His Leu Arg Thr His Thr Gly Glu Lys Pro Phe Ala
345 350 355
tgt ggc gcc tgc ggc aag gcc ttc ggc cag agc tcc cag ctc atc cag 2179
Cys Gly Ala Cys Gly Lys Ala Phe Gly Gln Ser Ser Gln Leu Ile Gln
360 365 370
cac cag cgg gtg cac tac cgc gag tagccgggcg ggggctcggg gctcggcctt 2233
His Gln Arg Val His Tyr Arg Glu
375 380
cctacctgcc cccaacccac cctccacccc gtcccccacg gtgggcactg cccagcaccg 2293
catgccacgt gtccggaata aattcttttt gattgttgga aaaaaaaaaa aaaaaaaa 2351
<210>9
<211>381
<212>PRT
＜213〉homo sapiens (Homo sapiens)
<400>9
Met Val Pro Ala Ala Leu Gly His Pro Ala Thr Pro Pro Thr Pro Val
1 5 10 15
Ser Met Ser Ala Leu Leu Gly Thr Ala Leu Arg Ala Pro Val Arg Leu
20 25 30
Ser Pro Ile His Trp Val Pro Gly Pro Ser Leu Ser Pro Leu Ser Pro
35 40 45
Pro Val Gln Leu Cys Ser Arg Ser Pro Ser Pro Ser Leu Arg Ala Thr
50 55 60
Val Gly Ala Cys Gly Tyr Tyr Pro Pro Leu Pro Phe Leu Ile Ser Asp
65 70 75 80
Ser Glu Pro Lys Pro Glu Gln Ala Pro Arg Ser Pro Gly Ser Gln Ala
85 90 95
Pro Asp Glu Gly Ala Gly Gly Ala Leu Arg Ser Leu Leu Arg Ser Leu
100 105 110
Pro Arg Arg Ala Arg Cys Ser Ala Gly Phe Gly Pro Glu Ser Ser Ala
115 120 125
Glu Arg Pro Ala Gly Gln Pro Pro Gly Ala Val Pro Cys Ala Gln Pro
130 135 140
Arg Gly Ala Trp Arg Val Thr Leu Val Gln Gln Ala Ala Ala Gly Pro
145 150 155 160
Glu Gly Ala Pro Glu Arg Ala Ala Glu Leu Gly Val Asn Phe Gly Arg
165 170 175
Ser Arg Gln Gly Ser Ala Arg Gly Thr Lys Pro His Arg Cys Glu Ala
180 185 190
Cys Gly Lys Ser Phe Lys Tyr Asn Ser Leu Leu Leu Lys His Gln Arg
195 200 205
Ile His Thr Gly Glu Lys Pro Tyr Ala Cys His Glu Cys Gly Lys Arg
210 215 220
Phe Arg Gly Trp Ser Gly Phe Ile Gln His His Arg Ile His Thr Gly
225 230 235 240
Glu Lys Pro Tyr Glu Cys Gly Gln Cys Gly Arg Ala Phe Ser His Ser
245 250 255
Ser His Phe Thr Gln His Leu Arg Ile His Asn Gly Glu Lys Pro Tyr
260 265 270
Lys Cys Gly Glu Cys Gly Gln Ala Phe Ser Gln Ser Ser Asn Leu Val
275 280 285
Arg His Gln Arg Leu His Thr Gly Glu Lys Pro Tyr Ala Cys Ser Gln
290 295 300
Cys Gly Lys Ala Phe Ile Trp Ser Ser Val Leu Ile Glu His Gln Arg
305 310 315 320
Ile His Thr Gly Glu Lys Pro Tyr Glu Cys Ser Asp Cys Gly Lys Ala
325 330 335
Phe Arg Gly Arg Ser His Phe Phe Arg His Leu Arg Thr His Thr Gly
340 345 350
Glu Lys Pro Phe Ala Cys Gly Ala Cys Gly Lys Ala Phe Gly Gln Ser
355 360 365
Ser Gln Leu Ile Gln His Gln Arg Val His Tyr Arg Glu
370 375 380
<210>10
<211>1413
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<400>10
gatggtttct tcagctgtca aatctcaggg tgctgggagt actgaataaa ttaaggcatg 60
ttaggtcagt ggggcagcgc ctggcagctg ggaaccaact ttattcccct cctctccttg 120
aaaagctctc ataattggcc tactccatcc cgcccactct tggtcactct cacttggtct 180
ttccttccaa gtcatcaagt gatttccaga gccactgcat ggaccttcct gagatggtgg 240
caagctgcct cagtacacac attaagccag catttaaaaa ggcactgcca gccgagcgtg 300
gtacctcata cctgtaatcc cagcactttg ggaggccgag gtgggcggat cccttcaggt 360
caggagtgca agaccagcct gaccttcatg gcaaaacccc gtctctacta aaaatacgaa 420
aattagccag gcatggtggc acacacctgg aggaggctct gtctcaaaaa aaaaaaaaaa 480
agaggcactg ccttggagaa gctgtcctga tcagcagcac atcttagact gtgggattac 540
aggttccaca gtactgggca gaaaccacct agagtaggcc aggcgcagtg gctcacgcct 600
gtaatcccag cactttggga agcccaggta ggcagatcac ttgaggccag gctggtctcg 660
gacacctgac ctcaggtgct gatcactttt tttatataat cttgaattgc attaaaggtt 720
tcctcatagg aaaccatatc aaagggacct ttgatagcag agaatgttct ttttaggctc 780
cccaaccacc tcaagtaccc tgaagcatca ttcccaattt taagagagtg tttatcttca 840
tttaaactct gtaatggttt ctctgtaatc acaaggatcc tgctccagac tgtcaccctc 900
ccagccttgc caggatgtcc agatgagcag tggtgtggag tcactagggg cacacattct 960
agagtcagac agcttgggtt tgaatcctga ctcccctgct aattcagcag ataacccagg 1020
gtgtctcaca caatctctcc agctcttgct ttaaaaataa aaaagcccgt ccatatagag 1080
gatgggtgaa taaatggtgg tatatctacc ccattaatgc agttggaatt agagagagat 1140
tttccatggg gtacctacag tttaatgaaa aatgcgagat gcagggaaaa atcgtgaaga 1200
tgtatgattc catttttgtc aaacaatgac ccaaaagcca tgaatgcaaa cgtgtgtgca 1260
caggattata caagtagaca gaaaaatatg gaagggtata tactcctaac acagggtacc 1320
aggatgcagg ggaagaaggg aggatggcaa cagcagggag ggcaagagag agaattggga 1380
aggaaagagc caagcaaaaa aaaaaaaaaa aaa 1413
<210>11
<211>1413
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<220>
<221>CDS
<222>(58)..(408)
<223>
<400>11
gatggtttct tcagctgtca aatctcaggg tgctgggagt actgaataaa ttaaggc 57
atg tta ggt cag tgg ggc agc gcc tgg cag ctg gga acc aac ttt att 105
Met Leu Gly Gln Trp Gly Ser Ala Trp Gln Leu Gly Thr Asn Phe Ile
1 5 10 15
ccc ctc ctc tcc ttg aaa agc tct cat aat tgg cct act cca tcc cgc 153
Pro Leu Leu Ser Leu Lys Ser Ser His Asn Trp Pro Thr Pro Ser Arg
20 25 30
cca ctc ttg gtc act ctc act tgg tct ttc ctt cca agt cat caa gtg 201
Pro Leu Leu Val Thr Leu Thr Trp Ser Phe Leu Pro Ser His Gln Val
35 40 45
att tcc aga gcc act gca tgg acc ttc ctg aga tgg tgg caa gct gcc 249
Ile Ser Arg Ala Thr Ala Trp Thr Phe Leu Arg Trp Trp Gln Ala Ala
50 55 60
tca gta cac aca tta agc cag cat tta aaa agg cac tgc cag ccg agc 297
Ser Val His Thr Leu Ser Gln His Leu Lys Arg His Cys Gln Pro Ser
65 70 75 80
gtg gta cct cat acc tgt aat ccc agc act ttg gga ggc cga ggt ggg 345
Val Val Pro His Thr Cys Asn Pro Ser Thr Leu Gly Gly Arg Gly Gly
85 90 95
cgg atc cct tca ggt cag gag tgc aag acc agc ctg acc ttc atg gca 393
Arg Ile Pro Ser Gly Gln Glu Cys Lys Thr Ser Leu Thr Phe Met Ala
100 105 110
aaa ccc cgt ctc tac taaaaatacg aaaattagcc aggcatggtg gcacacacct 448
Lys Pro Arg Leu Tyr
115
ggaggaggct ctgtctcaaa aaaaaaaaaa aaagaggcac tgccttggag aagctgtcct 508
gatcagcagc acatcttaga ctgtgggatt acaggttcca cagtactggg cagaaaccac 568
ctagagtagg ccaggcgcag tggctcacgc ctgtaatccc agcactttgg gaagcccagg 628
taggcagatc acttgaggcc aggctggtct cggacacctg acctcaggtg ctgatcactt 688
tttttatata atcttgaatt gcattaaagg tttcctcata ggaaaccata tcaaagggac 748
ctttgatagc agagaatgtt ctttttaggc tccccaacca cctcaagtac cctgaagcat 808
cattcccaat tttaagagag tgtttatctt catttaaact ctgtaatggt ttctctgtaa 868
tcacaaggat cctgctccag actgtcaccc tcccagcctt gccaggatgt ccagatgagc 928
agtggtgtgg agtcactagg ggcacacatt ctagagtcag acagcttggg tttgaatcct 988
gactcccctg ctaattcagc agataaccca gggtgtctca cacaatctct ccagctcttg 1048
ctttaaaaat aaaaaagccc gtccatatag aggatgggtg aataaatggt ggtatatcta 1108
ccccattaat gcagttggaa ttagagagag attttccatg gggtacctac agtttaatga 1168
aaaatgcgag atgcagggaa aaatcgtgaa gatgtatgat tccatttttg tcaaacaatg 1228
acccaaaagc catgaatgca aacgtgtgtg cacaggatta tacaagtaga cagaaaaata 1288
tggaagggta tatactccta acacagggta ccaggatgca ggggaagaag ggaggatggc 1348
aacagcaggg agggcaagag agagaattgg gaaggaaaga gccaagcaaa aaaaaaaaaa 1408
aaaaa 1413
<210>12
<211>117
<212>PRT
＜213〉homo sapiens (Homo sapiens)
<400>12
Met Leu Gly Gln Trp Gly Ser Ala Trp Gln Leu Gly Thr Asn Phe Ile
1 5 10 15
Pro Leu Leu Ser Leu Lys Ser Ser His Asn Trp Pro Thr Pro Ser Arg
20 25 30
Pro Leu Leu Val Thr Leu Thr Trp Ser Phe Leu Pro Ser His Gln Val
35 40 45
Ile Ser Arg Ala Thr Ala Trp Thr Phe Leu Arg Trp Trp Gln Ala Ala
50 55 60
Ser Val His Thr Leu Ser Gln His Leu Lys Arg His Cys Gln Pro Ser
65 70 75 80
Val Val Pro His Thr Cys Asn Pro Ser Thr Leu Gly Gly Arg Gly Gly
85 90 95
Arg Ile Pro Ser Gly Gln Glu Cys Lys Thr Ser Leu Thr Phe Met Ala
100 105 110
Lys Pro Arg Leu Tyr
115
<210>13
<211>2055
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<400>13
gcatggacga ccgagggcgg cactgctcac aggacggcta cccatccgca atggcttcta 60
caccaccaac gcccatgcca gaaacgccta cacaccgcag agattgtggg cggcatccca 120
gactcggagc agctcctgcc ggagcttctg aagaaggccg gctacgtcag caagattgtc 180
ggcaagtggc atctgggtca caggccccag ttccaccccc tgaagcacgg atttgatgag 240
tggtttggat cccccaactg ccactttgga ccttatgaca acaaggccag gcccaacatc 300
cctgtgtaca gggactggga gatggttggc agatattatg aagaatttcc tattaatctg 360
aagacggggg aagccaacct cacccagatc tacctgcagg aagccctgga cttcattaag 420
agacaggcac ggcaccaccc ctttttcctc tactgggctg tcgacgccac gcacgcaccc 480
gtctatgcct ccaaaccctt cttgggcacc agtcagcgag ggcggtatgg agacgccgtc 540
cgggagattg atgacagcat tgggaagata ctggagctcc tccaagacct gcacgtcgcg 600
gacaacacct tcgtcttctt cacgtcggac aacggcgctg ccctcatttc cgcccccgaa 660
caaggtggca gcaacggccc ctttctgtgt gggaagcaga ccacgtttga aggagggatg 720
agggagcctg ccctcgcatg gtggccaggg cacgtcactg caggccaggt gagccaccag 780
ctgggcagca tcatggacct cttcaccacc agcctggccc ttgcgggcct gacgccgccc 840
agcgacaggg ccattgatgg cctcaacctc ctccccaccc tcctgcaggg ccggctgatg 900
gacaggccta tcttctatta ccgtggcgac acgctgatgg cggccaccct cgggcagcac 960
aaggctcact tctggacctg gaccaactcc tgggagaact tcagacaggg cattgatttc 1020
tgccctgggc agaacgtttc aggggtcaca actcacaatc tggaagacca cacgaagctg 1080
cccctgatct tccacctggg acgggaccca ggggagaggt tccccctcag ctttgccagc 1140
gccgagtacc aggaggccct cagcaggatc acctcggtcg tccagcagca ccaggaggcc 1200
ttggtccccg cgcagcccca gctcaacgtg tgcaactggg cggtcatgaa ctgggcacct 1260
ccgggctgtg aaaagttagg gaagtgtctg acacctccag aatccattcc caagaagtgc 1320
ctctggtccc actagcacct gcgcagactc aggccaggcc tagaatctcc ggttggccct 1380
gcaagtgcct ggaggaagga tggctctggc ctcggtcctc ccccaaccct gcccaagcca 1440
gacagacagc acctgcagac gcagggggac tgcacaattc cacctgccca ggacctgacc 1500
ctggcgtgtg cttggccctc ctcctcgccc acggcgcctc agatttcagg accctcctcc 1560
tcgcccacgg cgcctcagac ctcaggaccc tgccgtctca cgcctttgtg aaccccaaat 1620
atctgagacc agtctcagtt tattttgcca aggttaagga tgcacctgtg acagcctcag 1680
gaggtcctga caacaggtgc ctgaggtggc tggggataca gtttgccttt atacatctta 1740
gggagacaca agatcagtat gtgtatggcg tacattggtt cagtcagcct tccactgaat 1800
acacgattga gtctggccca gtgaatccgc atttttatgt aaacagtaag ggaacggggc 1860
aatcatataa gcgtttgtct caggggagcc ccagagggat gacttccagt tccgtctgtc 1920
ctttgtccac aaggaatttc cctggacgct aattatgagg gaggcgtgta gcttcttatc 1980
attgtaacta tgttatttag aaataaaacg ggaggcaggt ttcctaattc ccagcttgaa 2040
aaaaaaaaaa aaaaa 2055
<210>14
<211>2055
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<220>
<221>CDS
<222>(322)..(1332)
<223>
<400>14
gcatggacga ccgagggcgg cactgctcac aggacggcta cccatccgca atggcttcta 60
caccaccaac gcccatgcca gaaacgccta cacaccgcag agattgtggg cggcatccca 120
gactcggagc agctcctgcc ggagcttctg aagaaggccg gctacgtcag caagattgtc 180
ggcaagtggc atctgggtca caggccccag ttccaccccc tgaagcacgg atttgatgag 240
tggtttggat cccccaactg ccactttgga ccttatgaca acaaggccag gcccaacatc 300
cctgtgtaca gggactggga g atg gtt ggc aga tat tat gaa gaa ttt cct 351
Met Val Gly Arg Tyr Tyr Glu Glu Phe Pro
1 5 10
att aat ctg aag acg ggg gaa gcc aac ctc acc cag atc tac ctg cag 399
Ile Asn Leu Lys Thr Gly Glu Ala Asn Leu Thr Gln Ile Tyr Leu Gln
15 20 25
gaa gcc ctg gac ttc att aag aga cag gca cgg cac cac ccc ttt ttc 447
Glu Ala Leu Asp Phe Ile Lys Arg Gln Ala Arg His His Pro Phe Phe
30 35 40
ctc tac tgg gct gtc gac gcc acg cac gca ccc gtc tat gcc tcc aaa 495
Leu Tyr Trp Ala Val Asp Ala Thr His Ala Pro Val Tyr Ala Ser Lys
45 50 55
ccc ttc ttg ggc acc agt cag cga ggg cgg tat gga gac gcc gtc cgg 543
Pro Phe Leu Gly Thr Ser Gln Arg Gly Arg Tyr Gly Asp Ala Val Arg
60 65 70
gag att gat gac agc att ggg aag ata ctg gag ctc ctc caa gac ctg 591
Glu Ile Asp Asp Ser Ile Gly Lys Ile Leu Glu Leu Leu Gln Asp Leu
75 80 85 90
cac gtc gcg gac aac acc ttc gtc ttc ttc acg tcg gac aac ggc gct 639
His Val Ala Asp Asn Thr Phe Val Phe Phe Thr Ser Asp Asn Gly Ala
95 100 105
gcc ctc att tcc gcc ccc gaa caa ggt ggc agc aac ggc ccc ttt ctg 687
Ala Leu Ile Ser Ala Pro Glu Gln Gly Gly Ser Asn Gly Pro Phe Leu
110 115 120
tgt ggg aag cag acc acg ttt gaa gga ggg atg agg gag cct gcc ctc 735
Cys Gly Lys Gln Thr Thr Phe Glu Gly Gly Met Arg Glu Pro Ala Leu
125 130 135
gca tgg tgg cca ggg cac gtc act gca ggc cag gtg agc cac cag ctg 783
Ala Trp Trp Pro Gly His Val Thr Ala Gly Gln Val Ser His Gln Leu
140 145 150
ggc agc atc atg gac ctc ttc acc acc agc ctg gcc ctt gcg ggc ctg 831
Gly Ser Ile Met Asp Leu Phe Thr Thr Ser Leu Ala Leu Ala Gly Leu
155 160 165 170
acg ccg ccc agc gac agg gcc att gat ggc ctc aac ctc ctc ccc acc 879
Thr Pro Pro Ser Asp Arg Ala Ile Asp Gly Leu Asn Leu Leu Pro Thr
175 180 185
ctc ctg cag ggc cgg ctg atg gac agg cct atc ttc tat tac cgt ggc 927
Leu Leu Gln Gly Arg Leu Met Asp Arg Pro Ile Phe Tyr Tyr Arg Gly
190 195 200
gac acg ctg atg gcg gcc acc ctc ggg cag cac aag gct cac ttc tgg 975
Asp Thr Leu Met Ala Ala Thr Leu Gly Gln His Lys Ala His Phe Trp
205 210 215
acc tgg acc aac tcc tgg gag aac ttc aga cag ggc att gat ttc tgc 1023
Thr Trp Thr Asn Ser Trp Glu Asn Phe Arg Gln Gly Ile Asp Phe Cys
220 225 230
cct ggg cag aac gtt tca ggg gtc aca act cac aat ctg gaa gac cac 1071
Pro Gly Gln Asn Val Ser Gly Val Thr Thr His Asn Leu Glu Asp His
235 240 245 250
acg aag ctg ccc ctg atc ttc cac ctg gga cgg gac cca ggg gag agg 1119
Thr Lys Leu Pro Leu Ile Phe His Leu Gly Arg Asp Pro Gly Glu Arg
255 260 265
ttc ccc ctc agc ttt gcc agc gcc gag tac cag gag gcc ctc agc agg 1167
Phe Pro Leu Ser Phe Ala Ser Ala Glu Tyr Gln Glu Ala Leu Ser Arg
270 275 280
atc acc tcg gtc gtc cag cag cac cag gag gcc ttg gtc ccc gcg cag 1215
Ile Thr Ser Val Val Gln Gln His Gln Glu Ala Leu Val Pro Ala Gln
285 290 295
ccc cag ctc aac gtg tgc aac tgg gcg gtc atg aac tgg gca cct ccg 1263
Pro Gln Leu Asn Val Cys Asn Trp Ala Val Met Asn Trp Ala Pro Pro
300 305 310
ggc tgt gaa aag tta ggg aag tgt ctg aca cct cca gaa tcc att ccc 1311
Gly Cys Glu Lys Leu Gly Lys Cys Leu Thr Pro Pro Glu Ser Ile Pro
315 320 325 330
aag aag tgc ctc tgg tcc cac tagcacctgc gcagactcag gccaggccta 1362
Lys Lys Cys Leu Trp Ser His
335
gaatctccgg ttggccctgc aagtgcctgg aggaaggatg gctctggcct cggtcctccc 1422
ccaaccctgc ccaagccaga cagacagcac ctgcagacgc agggggactg cacaattcca 1482
cctgcccagg acctgaccct ggcgtgtgct tggccctcct cctcgcccac ggcgcctcag 1542
atttcaggac cctcctcctc gcccacggcg cctcagacct caggaccctg ccgtctcacg 1602
cctttgtgaa ccccaaatat ctgagaccag tctcagttta ttttgccaag gttaaggatg 1662
cacctgtgac agcctcagga ggtcctgaca acaggtgcct gaggtggctg gggatacagt 1722
ttgcctttat acatcttagg gagacacaag atcagtatgt gtatggcgta cattggttca 1782
gtcagccttc cactgaatac acgattgagt ctggcccagt gaatccgcat ttttatgtaa 1842
acagtaaggg aacggggcaa tcatataagc gtttgtctca ggggagcccc agagggatga 1902
cttccagttc cgtctgtcct ttgtccacaa ggaatttccc tggacgctaa ttatgaggga 1962
ggcgtgtagc ttcttatcat tgtaactatg ttatttagaa ataaaacggg aggcaggttt 2022
cctaattccc agcttgaaaa aaaaaaaaaa aaa 2055
<210>15
<211>337
<212>PRT
＜213〉homo sapiens (Homo sapiens)
<400>15
Met Val Gly Arg Tyr Tyr Glu Glu Phe Pro Ile Asn Leu Lys Thr Gly
1 5 10 15
Glu Ala Asn Leu Thr Gln Ile Tyr Leu Gln Glu Ala Leu Asp Phe Ile
20 25 30
Lys Arg Gln Ala Arg His His Pro Phe Phe Leu Tyr Trp Ala Val Asp
35 40 45
Ala Thr His Ala Pro Val Tyr Ala Ser Lys Pro Phe Leu Gly Thr Ser
50 55 60
Gln Arg Gly Arg Tyr Gly Asp Ala Val Arg Glu Ile Asp Asp Ser Ile
65 70 75 80
Gly Lys Ile Leu Glu Leu Leu Gln Asp Leu His Val Ala Asp Asn Thr
85 90 95
Phe Val Phe Phe Thr Ser Asp Asn Gly Ala Ala Leu Ile Ser Ala Pro
100 105 110
Glu Gln Gly Gly Ser Asn Gly Pro Phe Leu Cys Gly Lys Gln Thr Thr
115 120 125
Phe Glu Gly Gly Met Arg Glu Pro Ala Leu Ala Trp Trp Pro Gly His
130 135 140
Val Thr Ala Gly Gln Val Ser His Gln Leu Gly Ser Ile Met Asp Leu
145 150 155 160
Phe Thr Thr Ser Leu Ala Leu Ala Gly Leu Thr Pro Pro Ser Asp Arg
165 170 175
Ala Ile Asp Gly Leu Asn Leu Leu Pro Thr Leu Leu Gln Gly Arg Leu
180 185 190
Met Asp Arg Pro Ile Phe Tyr Tyr Arg Gly Asp Thr Leu Met Ala Ala
195 200 205
Thr Leu Gly Gln His Lys Ala His Phe Trp Thr Trp Thr Asn Ser Trp
210 215 220
Glu Asn Phe Arg Gln Gly Ile Asp Phe Cys Pro Gly Gln Asn Val Ser
225 230 235 240
Gly Val Thr Thr His Asn Leu Glu Asp His Thr Lys Leu Pro Leu Ile
245 250 255
Phe His Leu Gly Arg Asp Pro Gly Glu Arg Phe Pro Leu Ser Phe Ala
260 265 270
Ser Ala Glu Tyr Gln Glu Ala Leu Ser Arg Ile Thr Ser Val Val Gln
275 280 285
Gln His Gln Glu Ala Leu Val Pro Ala Gln Pro Gln Leu Asn Val Cys
290 295 300
Asn Trp Ala Val Met Asn Trp Ala Pro Pro Gly Cys Glu Lys Leu Gly
305 310 315 320
Lys Cys Leu Thr Pro Pro Glu Ser Ile Pro Lys Lys Cys Leu Trp Ser
325 330 335
His
<210>16
<211>2392
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<400>16
gctgtactgt tggatttaac acccaccccg ctgaggcaag agggtcctgg cttagagatg 60
gcaaagcccg ccttctgggc agggcagaag gtggctgtgc agcctcctgg agctccagca 120
ggaccggcag agccctgagt cagcctcgta aattgtgtga caggcaagac cagaaagggc 180
tgatgttgaa acctggagag gagccctgca ggagctctcc aggagggagg gccctctcct 240
aatgatgctg gcctgaaagg gcctgaggcc gaaaggtgag ggggcggggg ctgttcagga 300
cacctggtct gtgtactcac ggcctctatc taccctgttc aggggaaccc caaacctgct 360
tttctttctt tctcagcctc tcctcccttc tcacaccctg gatgctgaga gcagggccca 420
gctctgctct ttccacaaca tttgggccat gttggcacag agcgggggcc tcttgggcta 480
tggtggccac actccccggg ctaactgagt tacctgattc ccagcggaaa ggtcaggaag 540
ggagggaaga aacttgctga gctacaaatt cgagactgct ggggctttct aggacaggcc 600
accttcctgc aattgtccct ctgtgtcccc aactccccgc agtcccgcag gccaggaggt 660
gggtgcctga tgagcggtgc ttccctcaga tgatatgtgg gcaccctgaa gctctcacgt 720
aatggttctg ctgtgccggg ttgtgaccca gctgtccttg gtaggctctc ctgtttaatg 780
agcaactgct atatgccagg ccctgttcta gaaacagatg aggcccctgt tcccatggaa 840
cttagatctg agtatgtgga cagagtgagt aggttgccag ataatgtcag aggtaagaaa 900
aaagccaggc agaggacagc agtgctggtt tagacaaggg ttggcaaggc ctccctgata 960
aggggacgta tttgcgcaga cacggaagca gtctgatgga aggcttgccc tgtagggacc 1020
tggggtgcag actgagctgg gagggctctt ggcttgctcc aatgaggtag gggcgaagag 1080
agggcagaat ggcccccgtg ctggtccaga tgtggccagc gccagatcgt atagagccgt 1140
ggaggcacgg taaggacttg gggtttgagg gagatgaggg ccctgagcag gggtgccatg 1200
atctagcctc tgctccagaa ggatctgtct ggctgccatg tgggggacac ggagtggagc 1260
aaagacggga gcccagtgag agtcgaggga gaggtgacct cagcttgggc tgcggtgcag 1320
gccatgggac tggcaggagg tggctggggt tacgctgtct tttctaacat tcagcattgg 1380
ctcaggggcc aggtcagggc tcacagactg ttgctataaa gggctgggtg gctttcttcc 1440
ccacagctac tcagcctaat gccattgcag agcacatgta gccatggaca acacaagggg 1500
cgtatccgtg ttccaggaca gccatattga caggaatagg caggaggcca gatttggtcc 1560
tcaggctgta atttcttggc cccttgtcta gggagaggta aacgagggga ggagagatca 1620
gtcaaggatg acgtgagggt ttgctgggag caccaggaat cctggagaag gtagtggcaa 1680
gagggtgcag caagctcagc tgggcgggga tcaagtctga ggacttaatg tctcctctga 1740
tctccagacc cataagggag atgctgagta gacaactggg gcttatgggt ctggagttca 1800
gaggagagat cgggaaggtg tccatttgga gtcatccacg cagagatgtg tgaaggctgc 1860
tcaatgattt tgaggtttaa agaaaaaaag agatgtgaaa ccaggggccc tgatgaggct 1920
gcccaggtgg taaggaagac agaagagaag ccatgggaca gctgagcccg ggcaccctca 1980
agccttggag gcatgaagtt tggtggggat ctggcaaaga acacctggga gcagccagcg 2040
ggcagcagac cccagagtag cagggaagac aagcacttca aagaggcagc gtcagccagg 2100
ggcagtggct cagctgtaat cctagcactt tgggaggcca aggatggcag atcacctgag 2160
gttaggggtt cgagactagc ctggccaaca tggtgaaacc ctgtctctac taaaaataaa 2220
aaattagcca gccattggtg gtatgtgcct gtaatcccag ctactcgaga ggctgaggca 2280
ggagaatctc ttgaacccgg gaggcagagg ttgcagtgag ccgagatcat gccattgcac 2340
tccagcctgg gcaacaagag cgaaactccg tctcaaaaaa aaaaaaaaaa aa 2392
<210>17
<211>2392
<212>DNA
＜213〉homo sapiens (Homo sapiens)
<220>
<221>CDS
<222>(1089)..(1364)
<223>
<400>17
gctgtactgt tggatttaac acccaccccg ctgaggcaag agggtcctgg cttagagatg 60
gcaaagcccg ccttctgggc agggcagaag gtggctgtgc agcctcctgg agctccagca 120
ggaccggcag agccctgagt cagcctcgta aattgtgtga caggcaagac cagaaagggc 180
tgatgttgaa acctggagag gagccctgca ggagctctcc aggagggagg gccctctcct 240
aatgatgctg gcctgaaagg gcctgaggcc gaaaggtgag ggggcggggg ctgttcagga 300
cacctggtct gtgtactcac ggcctctatc taccctgttc aggggaaccc caaacctgct 360
tttctttctt tctcagcctc tcctcccttc tcacaccctg gatgctgaga gcagggccca 420
gctctgctct ttccacaaca tttgggccat gttggcacag agcgggggcc tcttgggcta 480
tggtggccac actccccggg ctaactgagt tacctgattc ccagcggaaa ggtcaggaag 540
ggagggaaga aacttgctga gctacaaatt cgagactgct ggggctttct aggacaggcc 600
accttcctgc aattgtccct ctgtgtcccc aactccccgc agtcccgcag gccaggaggt 660
gggtgcctga tgagcggtgc ttccctcaga tgatatgtgg gcaccctgaa gctctcacgt 720
aatggttctg ctgtgccggg ttgtgaccca gctgtccttg gtaggctctc ctgtttaatg 780
agcaactgct atatgccagg ccctgttcta gaaacagatg aggcccctgt tcccatggaa 840
cttagatctg agtatgtgga cagagtgagt aggttgccag ataatgtcag aggtaagaaa 900
aaagccaggc agaggacagc agtgctggtt tagacaaggg ttggcaaggc ctccctgata 960
aggggacgta tttgcgcaga cacggaagca gtctgatgga aggcttgccc tgtagggacc 1020
tggggtgcag actgagctgg gagggctctt ggcttgctcc aatgaggtag gggcgaagag 1080
agggcaga atg gcc ccc gtg ctg gtc cag atg tgg cca gcg cca gat cgt 1130
Met Ala Pro Val Leu Val Gln Met Trp Pro Ala Pro Asp Arg
1 5 10
ata gag ccg tgg agg cac ggt aag gac ttg ggg ttt gag gga gat gag 1178
Ile Glu Pro Trp Arg His Gly Lys Asp Leu Gly Phe Glu Gly Asp Glu
15 20 25 30
ggc cct gag cag ggg tgc cat gat cta gcc tct gct cca gaa gga tct 1226
Gly Pro Glu Gln Gly Cys His Asp Leu Ala Ser Ala Pro Glu Gly Ser
35 40 45
gtc tgg ctg cca tgt ggg gga cac gga gtg gag caa aga cgg gag ccc 1274
Val Trp Leu Pro Cys Gly Gly His Gly Val Glu Gln Arg Arg Glu Pro
50 55 60
agt gag agt cga ggg aga ggt gac ctc agc ttg ggc tgc ggt gca ggc 1322
Ser Glu Ser Arg Gly Arg Gly Asp Leu Ser Leu Gly Cys Gly Ala Gly
65 70 75
cat ggg act ggc agg agg tgg ctg ggg tta cgc tgt ctt ttc 1364
His Gly Thr Gly Arg Arg Trp Leu Gly Leu Arg Cys Leu Phe
80 85 90
taacattcag cattggctca ggggccaggt cagggctcac agactgttgc tataaagggc 1424
tgggtggctt tcttccccac agctactcag cctaatgcca ttgcagagca catgtagcca 1484
tggacaacac aaggggcgta tccgtgttcc aggacagcca tattgacagg aataggcagg 1544
aggccagatt tggtcctcag gctgtaattt cttggcccct tgtctaggga gaggtaaacg 1604
aggggaggag agatcagtca aggatgacgt gagggtttgc tgggagcacc aggaatcctg 1664
gagaaggtag tggcaagagg gtgcagcaag ctcagctggg cggggatcaa gtctgaggac 1724
ttaatgtctc ctctgatctc cagacccata agggagatgc tgagtagaca actggggctt 1784
atgggtctgg agttcagagg agagatcggg aaggtgtcca tttggagtca tccacgcaga 1844
gatgtgtgaa ggctgctcaa tgattttgag gtttaaagaa aaaaagagat gtgaaaccag 1904
gggccctgat gaggctgccc aggtggtaag gaagacagaa gagaagccat gggacagctg 1964
agcccgggca ccctcaagcc ttggaggcat gaagtttggt ggggatctgg caaagaacac 2024
ctgggagcag ccagcgggca gcagacccca gagtagcagg gaagacaagc acttcaaaga 2084
ggcagcgtca gccaggggca gtggctcagc tgtaatccta gcactttggg aggccaagga 2144
tggcagatca cctgaggtta ggggttcgag actagcctgg ccaacatggt gaaaccctgt 2204
ctctactaaa aataaaaaat tagccagcca ttggtggtat gtgcctgtaa tcccagctac 2264
tcgagaggct gaggcaggag aatctcttga acccgggagg cagaggttgc agtgagccga 2324
gatcatgcca ttgcactcca gcctgggcaa caagagcgaa actccgtctc aaaaaaaaaa 2384
aaaaaaaa 2392
<210>18
<211>92
<212>PRT
＜213〉homo sapiens (Homo sapiens)
<400>18
Met Ala Pro Val Leu Val Gln Met Trp Pro Ala Pro Asp Arg Ile Glu
1 5 10 15
Pro Trp Arg His Gly Lys Asp Leu Gly Phe Glu Gly Asp Glu Gly Pro
20 25 30
Glu Gln Gly Cys His Asp Leu Ala Ser Ala Pro Glu Gly Ser Val Trp
35 40 45
Leu Pro Cys Gly Gly His Gly Val Glu Gln Arg Arg Glu Pro Ser Glu
50 55 60
Ser Arg Gly Arg Gly Asp Leu Ser Leu Gly Cys Gly Ala Gly His Gly
65 70 75 80
Thr Gly Arg Arg Trp Leu Gly Leu Arg Cys Leu Phe
85 90
<210>19
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>19
atttcctgac ctgtgatctg cc 22
<210>20
<211>21
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>20
tcttaccgaa cttgggtctc c 21
<210>21
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>21
ttctaggtcc gtgctgttgt aa 22
<210>22
<211>19
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>22
cgtgtccgtg aaatcgggc 19
<210>23
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>23
ctccaagacg tagaggaagt gg 22
<210>24
<211>18
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>24
ccgagccgga aggatgga 18
<210>25
<211>21
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>25
ctgtcaaatc tcagggtgct g 21
<210>26
<211>18
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>26
ttgtcgtccc tcccgttc 18
<210>27
<211>18
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>27
ggcactgctc acaggacg 18
<210>28
<211>20
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>28
aggcagacag gaaacaggtg 20
<210>29
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>29
tgagtcagcc tcgtaaattg tg 22
<210>30
<211>21
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>30
ttgttctcgc tttgaggcag a 21

Claims (5)

1. isolating polynucleotide is characterized in that, it comprises a nucleotide sequence, and this nucleotide sequence is selected from down group:
(a) coding has the proteic polynucleotide of people of cancer suppressing function, and described albumen has following aminoacid sequence: SEQID NO:6;
(b) with polynucleotide (a) complementary polynucleotide.
2. polynucleotide as claimed in claim 1 is characterized in that, the amino acid sequence of polypeptide of this polynucleotide encoding is shown in SEQ ID NO:6.
3. polynucleotide as claimed in claim 1 is characterized in that, the sequence of these polynucleotide is selected from down group:
Coding region sequence or the full length sequence of SEQ ID NO:5.
4. a carrier is characterized in that, it contains the described polynucleotide of claim 1.
5. a genetically engineered host cell is characterized in that, it is a kind of host cell that is selected from down group:
(a) host cell that transforms or transduce with the described carrier of claim 4;
(b) host cell that transforms or transduce with the described polynucleotide of claim 1.
CNB021364001A 2002-08-07 2002-08-07 Novel human protein with cancer inhibiting function and its code sequence Expired - Fee Related CN100478354C (en)

Priority Applications (3)

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CNB021364001A CN100478354C (en) 2002-08-07 2002-08-07 Novel human protein with cancer inhibiting function and its code sequence
PCT/CN2003/000639 WO2004014946A1 (en) 2002-08-07 2003-08-07 A novel homo protein with cancer suppressing function and its coding sequence
AU2003255098A AU2003255098A1 (en) 2002-08-07 2003-08-07 A novel homo protein with cancer suppressing function and its coding sequence

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNB021364001A CN100478354C (en) 2002-08-07 2002-08-07 Novel human protein with cancer inhibiting function and its code sequence

Publications (2)

Publication Number Publication Date
CN1473849A CN1473849A (en) 2004-02-11
CN100478354C true CN100478354C (en) 2009-04-15

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Family Applications (1)

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Country Status (1)

Country Link
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Publication number Publication date
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