CN1309135A - Novel human protein able to suppress cancer cell growth and its coding sequence - Google Patents
Novel human protein able to suppress cancer cell growth and its coding sequence Download PDFInfo
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Abstract
A novel anticancer human protein, the polynucleotide for coding it, the process for preparing said polypeptide by recombination, the application of said polypeptide in treating diseases including cancer, the antagonist of resisting to said polypeptide and its medical function and the usage of said polynucleotide are disclosed.
Description
The invention belongs to biological technical field, specifically, the present invention relates to the proteic polynucleotide of people that new coding has 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.
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.
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, and it comprises the polypeptide of the aminoacid sequence with the group of being selected from down: SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20; 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:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20.
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:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ IDNO:14, SEQ ID NO:17, SEQ ID NO:20.More preferably, the sequence of these polynucleotide is selected from down group: coding region sequence or the full length sequence of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21.
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 a successive 10-800 Nucleotide in the above-mentioned polynucleotide.
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 disclosing of the technology of this paper is conspicuous to those skilled in the art.
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 to cancer cells (liver cancer cell), and its inhibiting rate is more than 50% or 50%.
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.Purity with protein polypeptide of cancer suppressing function can be used amino acid sequence analysis.
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.With SP329 albumen is example, and the coding region sequence of encoding mature polypeptide can be identical with the coding region sequence shown in the SEQ ID NO:3 or the varient of degeneracy.As used herein, " varient of degeneracy " is meant that in the present invention coding has the protein of SEQ ID NO:2, but with the differentiated nucleotide sequence of coding region sequence shown in the SEQ ID NO:3.Have the albumen of cancer suppressing function for 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 and activity with the mature polypeptide shown in the SEQ ID NO:2.
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.
In the above-mentioned method of mentioning, isolation of genomic DNA is least commonly used.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, ALaboratory Manual, 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.Science1985 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 (Science, 1984 of reorganization; 224:1431).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: and the expression vector based on T7 of in bacterium, expressing (Rosenberg, et al.Gene, 1987,56:125); The pMSXND expression vector of in mammalian cell, expressing (Lee and Nathans, J Bio Chem.263:3521,1988) 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 (Sambroook, et al.Molecular Cloning, a Laboratory Manual, cold Spring HarborLaboratory.New York, 1989) such as extracorporeal recombinant DNA technology, DNA synthetic technology, the interior recombinant technologys of body.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 PL promotor; 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, HSV thymidine kinase 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 aforesaid method 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.Its example 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, the albumen that can have a cancer suppressing function adds during bioanalysis measures, and determines 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 albumen with cancer suppressing function that the gene therapy vector (as virus vector) of reorganization can be designed to express variation is to suppress endogenic protein-active with cancer suppressing function.For example, a kind of albumen with cancer suppressing function of variation can be the albumen with cancer suppressing function that shortens, lacked signal conduction function territory, though can combine with the substrate in downstream, lacks signaling activity.Therefore 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 is found in existing document (Sambrook, et al.).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.
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.This radiolabeled antibody can be used as a kind of atraumatic diagnostic method and is used for the location of tumour cell and has judged whether transfer.
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.A kind of usual method is with sulfydryl linking agent such as SPDP, attacks the amino of antibody, by the exchange of disulfide linkage, toxin is incorporated on the antibody, and this hybrid antibody can be used for killing the cell of the people's protein positive with cancer suppressing function.
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.Pat No.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 people with cancer suppressing function 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).This sequence can be specifically at certain bar human chromosome particular location and and can with its hybridization.Now, have only chromosomal marker thing seldom to can be used for the marker chromosomes position based on actual sequence data (repetition polymorphism).According to the present invention, for these sequences are associated with disease related gene, its important the first step is positioned these dna sequence dnas 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, MendelianInheritancein Man (can by with the online acquisition of JohnsHopkins 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.Resolving power according to present physical mapping and assignment of genes gene mapping technology, being accurately positioned to the cDNA of the chromosomal region relevant with disease, can be a kind of (the supposing that 1 megabasse mapping resolving power and every 20kb are corresponding to a gene) between 50 to 500 potential Disease-causing genes.
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 Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.
The acquisition of embodiment 1:cDNA gene and the restraining effect that the cancer cells clone is formed
SP329 comes from and buys liver cDNA library from GIBCO BRL company (cat, No.10422-012), PP203, PP238, PP856, PP1065, PP1221, PP2250 come from according to a conventional method the human placenta cDNA library who makes up.Get the placenta tissue at 3,6,10 monthly ages, (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 TMXR cDNA library construction test kit (Seratagene company).Wherein ThermoScript II is used MMLV-RT-Superscript II (GIBCO BRL) instead, and reverse transcription reaction carries out at 42 ℃.Transform the XL10-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 Qiagen96 orifice plate plasmid extraction test kit, 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 hours, the limit is observed, the training liquid that G418 concentration does not wait is changed on the limit.After about 2～3 times, there is the clone to form up to the microscopy cell, counting.Find SP329, PP203, PP238, PP856, PP1065, PP1221, PP2250 and PP3898 have the cell clone formation effect (inhibiting rate is more than 50% or 50%) of inhibition, and the result is as shown in table 1 below.
CDNA clone's transfectional cell (7721) clone formation situation
| CDNA clones title | C DNA cloning number (three repetitions) | Empty carrier clone number (three repetitions) |
| ????SP329 ????PP203 ????PP238 ????PP856 ????PP1065 ????PP1221 ????PP2250 ????PP3898 | ????2????0????0 ????6????8????13 ????0????0????0 ????0????1????0 ????2????0????1 ????0????0????1 ????0????0????1 ????2????0????0 | ????18????24????23 ????27????31????26 ????20????17????16 ????30????13????15 ????32????39????37 ????30????20????22 ????32????31????34 ????33????34????38 |
The cDNA clone is adopted two deoxidation cessation method, on the ABI377 automatic dna 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.
To finding after SP329, PP203, PP238, PP856, PP1065, PP1221, PP2250, the analysis of PP3898 cDNA cloned sequence that gene is still imperfect, adopt the Clontech SMARTRACE cDNA of company amplification kit (Cat.No.K1811-1), design gene specific primer (as shown in table 2 below), by specification is operated, and obtains full-length clone.
Table 2
| Clone's title | Special primer 1 | Special primer 2 |
| SP329 | ?5′GCTTCGCTCAAGAAGAAAAAGGCAC?3′ | 5′AGGACCTCCCAACTGCATGCCTC?3′ |
| PP203 | ?5′ACCCCTGGGATCCAACTCTTTGGTG?3′ | 5′CTGGGTATAGGCCACAGCGATCCAA?3′ |
| PP238 | ?5′TGTTCTGCCGGGAGGAGAAGCAGTA?3′ | 5′CCGTTCTTCTTCTCCACCTGCTCCC?3′ |
| PP856 | ?5′CCAGGGTGACTCAGCTGTCACTCCA?3′ | 5′CCAGAACTTTCCGCAGACCTTGTGC?3′ |
| PP1065 | ?5′GGAAAGGGGCTAGCATGAAGGTCCA?3′ | 5′TATGTTGGGGTGGGAGGAGCTCTGA?3′ |
| PP1221 | ?5′TATGGCTTTCTTGCCAGGAGGGGTC?3′ | 5′CCCTGGGTAGAACGGGTGAAGGGAT?3′ |
| PP2250 | ?5′GTCTGGGTATCAGCTGTTGGGGCCT?3′ | 5′ACAAGGAGAGAGTGCGGCTGCTGAG?3′ |
| PP3898 | ?5′TCATCCAGCCGGTCACTTGACTTGA?3′ | 5′GCCACAGCTGGTAGTTGGACTTGCC?3′ |
Particularly, each clone is used following primer: PP203 clone universal primer mix (uPM) Long 5 ' CTAATACGACTCACTATAGGGCAAGCAGTGGTAACAACGCAGAGT, 3 ' nido universal primer (NUP) 5 ' AAGCAGTGGTAACAACGCAGAGT 3 ' pp203-NB 5 ' ACCCCTGGGATCCAACTCTTTGGTG 3 ' pp203-B 5 ' CTGGGTATAGGCCACAGCGATCCAA 3 '
The PP238 clone
Universal primer mix (UPM) Long 5 ' CTAATACGACTCACTATAGGGCAAGCAGTGGTAACAACGCAGAGT 3 "
Nido universal primer (NUP) 5 ' AAGCAGTGGTAACAACGCAGAGT 3 '
pp238-NB, 5 ' TGTTCTGCCGGGAGGAGAAGCAGTA, 3 ' pp238-B, 5 ' CCGTTCTTCTTCTCCACCTGCTCCC, 3 ' PP856 clone universal primer mix, (UPM), Long, 5 ' CTAATACGACTCACTATAGGGCAAGCAGTGGTAACAACGCAGAGT, 3 ' nido universal primer, (NUP), 5 ' AAGCAGTGGTAACAACGCAGAGT, 3 ' pp856-NB, 5 ' CCAGGGTGACTCAGCTGTCACTCCA, 3 ' pp856-B, 5 ' CCAGAACTTTCCGCAGACCTTGTGC, 3 ' PP1065 clone universal primer mix, (UPM), Long, 5 ' CTAATACGACTCACTATAGGGCAAGCAGTGGTAACAACGCAGAGT, 3 ' nido universal primer, (NUP), 5 ' AAGCAGTGGTAACAACGCAGAGT, 3 ' PP1065-B:, 5 ' GGAAAGGGGCTAGCATGAAGGTCCA, 3 ' PP1065-NB, 5 ' TATGTTGGGGTGGGAGGAGCTCTGA, 3 ' PP1221 clone is connected primer 1, (AP1), 5 ' CCATCCTAATACGACTCACTATAGGGC, 3 ' 27 are connected primer 2, (AP2), 5 ' ACTCACTATAGGGCTCGAGCGGC, 3 ', 23pp1221-C, 5 ' TATGGCTTTCTTGCCAGGAGGGGTC, 3 ', 25pp1221-NC, 5 ' CCCTGGGTAGAACGGGTGAAGGGAT, 3 ', 25pp2250-C, 5 ' ACAAGGAGAGAGTGCGGCTGCTGAG, 3 ' pp2250-D, 5 ' TGCATGCCACTTTCAGTCAACAGGA, 3 ' PP329 clone is connected primer 1, (AP1), 5 ' CCATCCTAATACGACTCACTATAGGGC, 3 ' 27 are connected primer 2, (AP2), 5 ' ACTCACTATAGGGCTCGAGCGGC, 3 ', 23sp329-B, 5 ' gcttcgctcaagaagaaaaaggcac, 3 ', 25sp329-NB, 5 ' aggacctcccaactgcatgcctc, 3 ', 23
Wherein, to using the clone of universal primer, obtain full-length gene by following operation.With human placenta mRNA is parent material, the X-B that obtains inferior respectively then UPM primer of cDNA. and gene specific by Clontech company SM ART RACE cDNA amplification kit (cat#1811-1) specification sheets carries out the first soft PCR, carry out second with the X-NB of NUP primer and gene specific again and take turns PCR, obtain gene fragment (annotate: X is the gene clone number).
Reaction conditions is: 94 ℃ of 3 ' one circulations
(first round) 94 ℃ of 1 ' 58 ℃ of 1 ' 72 ℃ of 2 ' 35 circulations
72 ℃ of 10 ' one circulations
(second takes turns) is the same, but annealing temperature is 60 ℃
To using the clone who is connected primer, with the Marathon-Ready people of Clonteoh company liver cDNA (cat#7407-1) is model, carry out first round PCR with Apl and X-C primer respectively, carry out second with Apl and X-NC again and take turns PCR acquisition gene fragment (annotate: X is the gene clone number).Reaction conditions is the same.
PP2250 is a model with people's placenta cDNA, PP2250-C, PP2250-D be primer by 94 ℃ of 3 ' 1 circulations, 94 ℃ 1 ' 60 ℃ 1 ' 1 70 ℃ of 2 ' 35 circulation, PCR is carried out in 72 ℃ of 10 ' one circulations, obtains gene fragment.
Embodiment 2:cDNA cloned sequence is analyzed
1．SP329
A: the nucleotide sequence (SEQ ID NO: 1) Length: 2360
TGTCAGTAAG TGGAAAAGGC AAGACTCCAC TTCGAAAGAG GTACAACTCC CATCAGATGG 60
GCCAGTCGAA GCAGTTTCCC CTCGAGGAAA GCAGCTGTGA GAAAGGCTGT CAGGTCACCA 120
GTGAGCAGAT CAAAGCCGAT ATGAAAGCAG CTAGGGATAT TCCTGAAAAG AAAAAAAACA 180
AGGATGTTTA TCCCAGCTGC AGCAGCACCA CCGCCAGCAC AGTGGGAAAC TCCAGCTCAC 240
ACAACACTGC TTCTCAAAGC CCCGACTTTG TAAGGACGGT GAACAGCGGC GGCTCTTCCG 300
AGCCTAGCCC TACAGAAGTG GATGTGTCCA GGCAGTGTGC CTGCTCCCCC GGTGGGTCAG 360
AGGACTCTGA GGCCATGGAG GAGGGAGATG CAGAGAGTTC TGTCTGCCCA GATGCTTGCT 420
GTCACAGGCC CCAGGAATTC CCAAAGGAGA ACTAGCAGGT GTTCTGATGA GGAACGTCCT 480
TCAACCAGCC GAGCCTGTGT TGTGAATGGC CCGGATGGTA CGAGATCCGC CTTTTCCTTT 540
AGGACTCTGC CACAAGGGGG GTCTTCAGGC CCAGCACATG ATGAGAGGAC TAATGGGAGT 600
GGCTCTGGGG CTACAGGTGA GGACAGGAGG GGGAGCTCCC AGCCTGAGAG TTGTGACGTG 660
CAGTCTAATG AAGACTACCC TCGGAGGCCC CTAACCAGGG CCAGGAGCAG ACTGTCCCAT 720
GTACTGCTGG TATCTGAGTC AGAAGTAGCC AAAACAAAGC CACGTCACGC CATGAAACGG 780
AAGCGGACAG CAGATAAATC CACTAGTACA AGTGATCCTG TGATCGAGGA TGACCATGTG 840
CAGGTTCTTG TATTAAAATC CAAGAATCTT GTTGGAGTCA CTATGACCAA TTGTGGAATC 900
ACAGATCTAG TGCTAAAAGA CTGTCCAAAG ATGATGTTCA TCCATGCTAC CAGGTGCAGG 960
GTACTAAAAC ATTTAAAGGT AGAAAATGCA CCAATTGTAA ACCGATTTGA CTATGCACAG 1020
TGCAAGAAAC TGAACATGGA TCAGGTACTA GACCAGATAC TAAGAATGCC ACCCGAGAGA 1080
AACCGCATCA TATACCTACG CCCAATGCAG CAGGTGGACA CTCTAACTTT GGAGCAGAAG 1140
CTATTTAGTG GTCCCTACCC CTATCACATC TGTATTATCC ATGAATTCAG TAACCCTCCC 1200
AATGTCCGGA ATAAGGTGCG CATTCGCAGC TGGATGGACA CTATAGCAAA CATCAATCAA 1260
GAGCTCATTA AATATGAATT CTTCCCTGAA GCCACTCGAA GTGAAGAAGA CTTAAAGAAA 1320
TACCCCAAGT ACCCCTGGGG GAGAGAAATC TATACTTTAG AAGGTGTTGT GGATGGAGCT 1380
CCATATTCCA TGATTTCTGA CTTCCCTTGG CTGAGGTCAT TACGAGCTGC AGAGCCCAAC 1440
AGCTTCGCTC GATACGACTT TGAAGACGAT GAAGAAAGCA CTATCTATGC TCCTAGAAGG 1500
AAAGGACAGC TGTCTGCAGA CATCTGTATG GAAACAATAG GAGAGGAAAT TTCAGAGATG 1560
CGTCAGATGA AGAAGGGTGT ATTTCAGCGA GTAGTGGCAA TTTTTATCCA CTATTGTGAT 1620
GTCAATGGAG AGCCAGTTGA AGATGACTAC ATTTAATTGG TCCCTCCTCC TTTCCAGCTA 1680
TTTTGTCAGA AAGCAAGTAG GGCCATCCAG CTGCCAGAGT GCTCCACAGG GACTTGAGGC 1740
ATGCAGTTGG GAGGTCCTGG CTCGGTTTGC TATATAGGGA ATATATAAGG AACATCGAAA 1800
TTGTATACAA AGATTTGTAC ATAAAAAATA TACAAAGACG CTTCCTAAAG TACCAACTTT 1860
ATATCATATG TTTATACAAT TTAATTTAAA AATTCATTTT AAGGAAGACA GATAATTTGA 1920
AAGACTTTTG TTTTTCTTGA CTTAATTCAT GAAGTATCAT TTTTTGACTG AGTCTCCATT 1980
TACTTCATTC TTAATGATTA TTGTCATCCC TTTAAATCTG TGCCTTTTTC TTCTTGAGCG 2040
AAGCTGTTTG AGTAAACCTG TTGAAGAGTG TTTGTGTCTT GTGTGCTTTT TTGTTGTTAT 2100
TAAAACACCA ACTAAACCTT ATAGTCAAGA CAAGGCTCTA TGTTTCTGTA CAAAGCTGTA 2160
GTTCTTTCTT AGTATTATAG TTGCCATGTT TCTTAAAATC AAGTAAAAAG ACTTATGAGC 2220
TTAAAAAAAA GTGAGTTTGA GAGGGAAATG GAAAAGTTTC CAGAGTATTT CTAGTAATTA 2280
TTTCCACATT GAATTGTGTA TATGCTTTAT CTTGAATATA AAATAAAAGT TTATTAAAAA 2340
CTTTAAAAAA AAAAAAAAAA 2360
B: the amino acid sequence (SEQ ID NO: 2) Length : 444
1 MCPGSVPAPP VGQRTLRPWR REMQRVLSAQ MLAVTGPRNS QRRTSRCSDE
51 ERPSTSRACV VNGPDGTRSA FSFRTLPQGG SSGPAHDERT NGSGSGATGE
101 DRRGSSQPES CDVQSNEDYP RRPLTRARSR LSHVLLVSES EVAKTKPRHA
151 MKRKRTADKS TSTSDPVIED DHVQVLVLKS KNLVGVTMTN CGITDLVLKD
201 CPKMMFIHAT RCRVLKHLKV ENAPIVNRFD YAQCKKLNMD QVLDQILRMP
251 PERNRIIYLR PMQQVDTLTL EQKLFSGPYP YHICIIHEFS NPPNVRNKVR
301 IRSWMDTIAN INQELIKYEF FPEATRSEED LKKYPKYPWG REIYTLEGVV
351 DGAPYSMISD FPWLRSLRAA EPNSFARYDF EDDEESTIYA PRRKGQLSAD
401 ICMETIGEEI SEMRQMKKGV FQRVVAIFIH YCDVNGEPVE DDYI
Clone : SP329 (SEQ ID NO: 3)
Start codon : 322 ATG termination codon : 1656TAA Protein Weight: 50647.03
1 TGT CAG TAA GTG GAA AAG GCA AGA CTC CAC TTC GAA AGA GGT ACA ACT 48
49 CCC ATC AGA TGG GCC AGT CGA AGC AGT TTC CCC TCG AGG AAA GCA GCT 96
97 GTG AGA AAG GCT GTC AGG TCA CCA GTG AGC AGA TCA AAG CCG ATA TGA 144
145 AAG CAG CTA GGG ATA TTC CTG AAA AGA AAA AAA ACA AGG ATG TTT ATC 192
193 CCA GCT GCA GCA GCA CCA CCG CCA GCA CAG TGG GAA ACT CCA GCT CAC 240
241 ACA ACA CTG CTT CTC AAA GCC CCG ACT TTG TAA GGA CGG TGA ACA GCG 288
289 GCG GCT CTT CCG AGC CTA GCC CTA CAG AAG TGG ATG TGT CCA GGC AGT 336
1 Met Cys Pro Gly Ser 5
337 GTG CCT GCT CCC CCG GTG GGT CAG AGG ACT CTG AGG CCA TGG AGG AGG 384
6 Val Pro Ala Pro Pro Val Gly Gln Arg Thr Leu Arg Pro Trp Arg Arg 21
385 GAG ATG CAG AGA GTT CTG TCT GCC CAG ATG CTT GCT GTC ACA GGC CCC 432
22 Glu Met Gln Arg Val Leu Ser Ala Gln Met Leu Ala Val Thr Gly Pro 37
433 AGG AAT TCC CAA AGG AGA ACT AGC AGG TGT TCT GAT GAG GAA CGT CCT 480
38 Arg Asn Ser Gln Arg Arg Thr Ser Arg Cys Ser Asp Glu Glu Arg Pro 53
481 TCA ACC AGC CGA GCC TGT GTT GTG AAT GGC CCG GAT GGT ACG AGA TCC 528
54 Ser Thr Ser Arg Ala Cys Val Val Asn Gly Pro Asp Gly Thr Arg Ser 69
529 GCC TTT TCC TTT AGG ACT CTG CCA CAA GGG GGG TCT TCA GGC CCA GCA 576
70 Ala Phe Ser Phe Arg Thr Leu Pro Gln Gly Gly Ser Ser Gly Pro Ala 85
577 CAT GAT GAG AGG ACT AAT GGG AGT GGC TCT GGG GCT ACA GGT GAG GAC 624
86 His Asp Glu Arg Thr Asn Gly Ser Gly Ser Gly Ala Thr Gly Glu Asp 101
625 AGG AGG GGG AGC TCC CAG CCT GAG AGT TGT GAC GTG CAG TCT AAT GAA 672
102 Arg Arg Gly Ser Ser Gln Pro Glu Ser Cys Asp Val Gln Ser Asn Glu 117
673 GAC TAC CCT CGG AGG CCC CTA ACC AGG GCC AGG AGC AGA CTG TCC CAT 720
118 Asp Tyr Pro Arg Arg Pro Leu Thr Arg Ala Arg Ser Arg Leu Ser His 133
721 GTA CTG CTG GTA TCT GAG TCA GAA GTA GCC AAA ACA AAG CCA CGT CAC 768
134 Val Leu Leu Val Ser Glu Ser Glu Val Ala Lys Thr Lys Pro Arg His 149
769 GCC ATG AAA CGG AAG CGG ACA GCA GAT AAA TCC ACT AGT ACA AGT GAT 816
150 Ala Met Lys Arg Lys Arg Thr Ala Asp Lys Ser Thr Ser Thr Ser Asp 165
817 CCT GTG ATC GAG GAT GAC CAT GTG CAG GTT CTT GTA TTA AAA TCC AAG 864
166 Pro Val Ile Glu Asp Asp His Val Gln Val Leu Val Leu Lys Ser Lys 181
865 AAT CTT GTT GGA GTC ACT ATG ACC AAT TGT GGA ATC ACA GAT CTA GTG 912
182 Asn Leu Val Gly Val Thr Met Thr Asn Cys Gly Ile Thr Asp Leu Val 197
913 CTA AAA GAC TGT CCA AAG ATG ATG TTC ATC CAT GCT ACC AGG TGC AGG 960
198 Leu Lys Asp Cys Pro Lys Met Met Phe Ile His Ala Thr Arg Cys Arg 213
961 GTA CTA AAA CAT TTA AAG GTA GAA AAT GCA CCA ATT GTA AAC CGA TTT 1008
214 Val Leu Lys His Leu Lys Val Glu Asn Ala Pro Ile Val Asn Arg Phe 229
1009 GAC TAT GCA CAG TGC AAG AAA CTG AAC ATG GAT CAG GTA CTA GAC CAG 1056
230 Asp Tyr Ala Gln Cys Lys Lys Leu Asn Met Asp Gln Val Leu Asp Gln 245
1057 ATA CTA AGA ATG CCA CCC GAG AGA AAC CGC ATC ATA TAC CTA CGC CCA 1104
246 Ile Leu Arg Met Pro Pro Glu Arg Asn Arg Ile Ile Tyr Leu Arg Pro 261
1105 ATG CAG CAG GTG GAC ACT CTA ACT TTG GAG CAG AAG CTA TTT AGT GGT 1152
262 Met Gln Gln Val Asp Thr Leu Thr Leu Glu Gln Lys Leu Phe Ser Gly 277
1153 CCC TAC CCC TAT CAC ATC TGT ATT ATC CAT GAA TTC AGT AAC CCT CCC 1200
278 Pro Tyr Pro Tyr His Ile Cys Ile Ile His Glu Phe Ser Asn Pro Pro 293
1201 AAT GTC CGG AAT AAG GTG CGC ATT CGC AGC TGG ATG GAC ACT ATA GCA 1248
294 Asn Val Arg Asn Lys Val Arg Ile Arg Ser Trp Met Asp Thr Ile Ala 309
1249 AAC ATC AAT CAA GAG CTC ATT AAA TAT GAA TTC TTC CCT GAA GCC ACT 1296
310 Asn Ile Asn Gln Glu Leu Ile Lys Tyr Glu Phe Phe Pro Glu Ala Thr 325
1297 CGA AGT GAA GAA GAC TTA AAG AAA TAC CCC AAG TAC CCC TGG GGG AGA 1344
326 Arg Ser Glu Glu Asp Leu Lys Lys Tyr Pro Lys Tyr Pro Trp Gly Arg 341
1345 GAA ATC TAT ACT TTA GAA GGT GTT GTG GAT GGA GCT CCA TAT TCC ATG 1392
342 Glu Ile Tyr Thr Leu Glu Gly Val Val Asp Gly Ala Pro Tyr Ser Met 357
1393 ATT TCT GAC TTC CCT TGG CTG AGG TCA TTA CGA GCT GCA GAG CCC AAC 1440
358 Ile Ser Asp Phe Pro Trp Leu Arg Ser Leu Arg Ala Ala Glu Pro Asn 373
1441 AGC TTC GCT CGA TAC GAC TTT GAA GAC GAT GAA GAA AGC ACT ATC TAT 1488
374 Ser Phe Ala Arg Tyr Asp Phe Glu Asp Asp Glu Glu Ser Thr Ile Tyr 389
1489 GCT CCT AGA AGG AAA GGA CAG CTG TCT GCA GAC ATC TGT ATG GAA ACA 1536
390 Ala Pro Arg Arg Lys Gly Gln Leu Ser Ala Asp Ile Cys Met Glu Thr 405
1537 ATA GGA GAG GAA ATT TCA GAG ATG CGT CAG ATG AAG AAG GGT GTA TTT 1584
406 Ile Gly Glu Glu Ile Ser Glu Met Arg Gln Met Lys Lys Gly Val Phe 421
1585 CAG CGA GTA GTG GCA ATT TTT ATC CAC TAT TGT GAT GTC AAT GGA GAG 1632
422 Gln Arg Val Val Ala Ile Phe Ile His Tyr Cys Asp Val Asn Gly Glu 437
1633 CCA GTT GAA GAT GAC TAC ATT TAA TTG GTC CCT CCT CCT TTC CAG CTA 1680
438 Pro Val Glu Asp Asp Tyr Ile *** 445
1681 TTT TGT CAG AAA GCA AGT AGG GCC ATC CAG CTG CCA GAG TGC TCC ACA 1728
1729 GGG ACT TGA GGC ATG CAG TTG GGA GGT CCT GGC TCG GTT TGC TAT ATA 1776
1777 GGG AAT ATA TAA GGA ACA TCG AAA TTG TAT ACA AAG ATT TGT ACA TAA 1824
1825 AAA ATA TAC AAA GAC GCT TCC TAA AGT ACC AAC TTT ATA TCA TAT GTT 1872
1873 TAT ACA ATT TAA TTT AAA AAT TCA TTT TAA GGA AGA CAG ATA ATT TGA 1920
1921 AAG ACT TTT GTT TTT CTT GAC TTA ATT CAT GAA GTA TCA TTT TTT GAC 1968
1969 TGA GTC TCC ATT TAC TTC ATT CTT AAT GAT TAT TGT CAT CCC TTT AAA 2016
2017 TCT GTG CCT TTT TCT TCT TGA GCG AAG CTG TTT GAG TAA ACC TGT TGA 2064
2065 AGA GTG TTT GTG TCT TGT GTG CTT TTT TGT TGT TAT TAA AAC ACC AAC 2112
2113 TAA ACC TTA TAG TCA AGA CAA GGC TCT ATG TTT CTG TAC AAA GCT GTA 2160
2161 GTT CTT TCT TAG TAT TAT AGT TGC CAT GTT TCT TAA AAT CAA GTA AAA 2208
2209 AGA CTT ATG AGC TTA AAA AAA AGT GAG TTT GAG AGG GAA ATG GAA AAG 2256
2257 TTT CCA GAG TAT TTC TAG TAA TTA TTT CCA CAT TGA ATT GTG TAT ATG 2304
2305 CTT TAT CTT GAA TAT AAA ATA AAA GTT TAT TAA AAA CTT TAA AAA AAA 2352
2353 AAA AAA AA 2360
2．PP203
A: the nucleotide sequence (SEQ ID NO: 4) Length: 2379
GGTGCACACC CGGAAGTGGG TGCGGGCCAG CCGGCTCGCC CGGGGGCCAT GGCAGCAGCG 60
GCTACTGCAG CCGAGGGGGT CCCCAGTCGG GGGCCTCCCG GGGAAGTCAT TCATCTGAAT 120
GTGGGAGGCA AGAGATTCAG TACCTCTCGC CAGACTCTCA CCTGGATCCC AGACTCCTTC 180
TTCTCCAGTC TTCTGAGCGG ACGCATCTCG ACGCTGAAAG ATGAGACCGG AGCAATCTTC 240
ATCGACAGGG ACCCTACAGT CTTCGCCCCC ATCCTCAACT TCCTGCGCAC CAAAGAGTTG 300
GATCCCAGGG GTGTCCACGG TTCCAGCCTC CTCCATGAAG CCCAGTTCTA TGGGCTCACT 360
CCTCTGGTTC GTCGCCTGCA GCTTCGAGAG GAGTTGGATC GATCTTCTTG TGGAAACGTC 420
CTCTTCAATG GTTACCTGCC GCCACCAGTG TTCCCAGTGA AGCGGCGGAA CCGGCACAGC 480
CTAGTGGGGC CTCAGCAGCT AGGAGGACGG CCAGCCCCTG TCCGACGGAG CAACACGATG 540
CCCCCCAACC TTGGCAATGC AGGGCTGCTG GGCCGAATGC TGGATGAGAA AACCCCTCCC 600
TCACCCTCAG GACAACCTGA GGAGCCGGGG ATGGTGCGCC TGGTGTGTGG ACACCATAAT 660
TGGATCGCTG TGGCCTATAC CCAGTTTCTA GTCTGCTACA GGTTGAAGGA AGCCTCTGGC 720
TGGCAGCTGG TGTTTTCCAG CCCCCGCCTG GACTGGCCCA TCGAACGACT GGCGCTCACA 780
GCCCGGGTGC ATGGTGGGGC TTTGGGTGAA CATGACAAGA TGGTGGCAGC AGCCACCGGC 840
AGCGAGATCC TGCTATGGGC TCTGCAGGCG GAAGGCGGTG GCTCCGAGAT AGGGGTCTTT 900
CATCTGGGGG TGCCTGTGGA GGCCTTGTTC TTCGTCGGGA ACCAGCTCAT TGCTACAAGC 960
CACACAGGGC GCATCGGGGT GTGGAATGCC GTCACCAAGC ACTGGCAGGT CCAGGAGGTG 1020
CAGCCCATCA CCAGTTATGA CGCGGCAGGC TCCTTCCTCC TCCTGGGCTG CAACAACGGC 1080
TCCATTTACT ACGTGGATGT GCAGAAGTTC CCCTTGCGCA TGAAAGACAA CGACCTCCTT 1140
GTCAGCGAGC TCTATCGGGA CCCAGCGGAG GATGGGGTCA CCGCCCTCAG TGTCTACCTC 1200
ACCCCCAAGA CCAGTGACAG TGGGAACTGG ATCGAGATCG CCTATGGCAC CAGCTCAGGG 1260
GGCGTGCGGG TCATCGTGCA GCACCCGGAG ACTGTGGGCT CGGGGCCTCA GCTCTTCCAG 1320
ACCTTCACTG TGCACCGCAG CCCTGTCACC AAGATCATGC TGTCGGAGAA GCACCTCATC 1380
TCAGTCTGTG CCGACAACAA CCACGTGCGG ACATGGTCTG TGACTCGCTT CCGCGGCATG 1440
ATTTCCACCC AGCCCGGCTC CACCCCACTC GCTTCCTTTA AGATCCTGGC TCTGGAGTCG 1500
GCAGATGGGC ATGGCGGCTG CAGTGCTGGC AATGACATTG GCCCCTACGG TGAGCGGGAC 1560
GACCAGCAAG TGTTCATCCA GAAGGTGGTG CCCAGTGCCA GCCAGCTCTT CGTGCGTCTC 1620
TCATCTACTG GGCAGCGGGT GTGCTCCGTG CGCTCCGTGG ACGGCTCACC CACGACGGCC 1680
TTCACAGTGC TGGAGTGCGA GGGCTCCCGG CGGCTCGGCT CTCGGCCCCG GCGCTACCTG 1740
CTCACTGGCC AGGCCAACGG CAGCTTGGCC ATGTGGGACC TAACCACCGC CATGGACGGC 1800
CTCGGCCAGG CCCCTGCAGG TGGCCTGACG GAGCAAGAGC TGATGGAACA GCTGGAACAC 1860
TGTGAGCTGG CCCCGCCGGC TCCTTCAGCT CCCTCATGGG GCTGTCTCCC CAGCCCCTCA 1920
CCCCGCATCT CCCTCACCAG CCTCCACTCA GCCTCCAGCA ACACCTCCTT GTCTGGCCAC 1980
CGTGGGAGCC CAAGCCCCCC GCAGGCTGAG GCCCGGCGCC GTGGTGGGGG CAGCTTTGTG 2040
GAACGCTGCC AGGAACTGGT GCGGAGTGGG CCAGACCTCC GACGGCCACC CACACCAGCC 2100
CCGTGGCCCT CCAGCGGTCT CGGCACTCCC CTCACACCTC CCAAGATGAA GCTCAATGAA 2160
ACTTCCTTTT GAACAACGCA GCTGCCATGA TGCCTTGGGA TGCCCTGGTC CTGGGGGACT 2220
CAGGTGCCTC CCTGATTCCT GTGGGAACCC CGGGTTCAGG GCCAGGGCCT CCTTGGAATA 2280
AATGGTTATT GTTACTAGGT CCCCACCTTC CCTCTTTTCT GGAAGCCAAA GTCAGCCTCC 2340
CCAATAAAGT CCTCACTGCC AAAAAAAAAA AAAAAAAAA 2379
B: the amino acid sequence (SEQ ID NO: 5) Length: 707
1 MAAAATAAEG VPSRGPPGEV IHLNVGGKRF STSRQTLTWI PDSFFSSLLS
51 GRISTLKDET GAIFIDRDPT VFAPILNFLR TKELDPRGVH GSSLLHEAQF
101 YGLTPLVRRL QLREELDRSS CGNVLFNGYL PPPVFPVKRR NRHSLVGPQQ
151 LGGRPAPVRR SNTMPPNLGN AGLLGRMLDE KTPPSPSGQP EEPGMVRLVC
201 GHHNWIAVAY TQFLVCYRLK EASGWQLVFS SPRLDWPIER LALTARVHGG
251 ALGEHDKMVA AATGSEILLW ALQAEGGGSE IGVFHLGVPV EALFFVGNQL
301 IATSHTGRIG VWNAVTKHWQ VQEVQPITSY DAAGSFLLLG CNNGSIYYVD
351 VQKFPLRMKD NDLLVSELYR DPAEDGVTAL SVYLTPKTSD SGNWIEIAYG
401 TSSGGVRVIV QHPETVGSGP QLFQTFTVHR SPVTKIMLSE KHLISVCADN
451 NHVRTWSVTR FRGMISTQPG STPLASFKIL ALESADGHGG CSAGNDIGPY
501 GERDDQQVFI QKVVPSASQL FVRLSSTGQR VCSVRSVDGS PTTAFTVLEC
551 EGSRRLGSRP RRYLLTGQAN GSLAMWDLTT AMDGLGQAPA GGLTEQELME
601 QLEHCELAPP APSAPSWGCL PSPSPRISLT SLHSASSNTS LSGHRGSPSP
651 PQAEARRRGG GSFVERCQEL VRSGPDLRRP PTPAPWPSSG LGTPLTPPKM
701 KLNETSF
Clone: PP203 (SEQ ID NO: 6)
Start codon: 49 ATG termination codon: 2172TGA Protein Weight: 76339.52
1 GGT GCA CAC CCG GAA GTG GGT GCG GGC CAG CCG GCT CGC CCG GGG GCC 48
49 ATG GCA GCA GCG GCT ACT GCA GCC GAG GGG GTC CCC AGT CGG GGG CCT 96
1 Met Ala Ala Ala Ala Thr Ala Ala Glu Gly Val Pro Ser Arg Gly Pro 16
97 CCC GGG GAA GTC ATT CAT CTG AAT GTG GGA GGC AAG AGA TTC AGT ACC 144
17 Pro Gly Glu Val Ile His Leu Asn Val Gly Gly Lys Arg Phe Ser Thr 32
145 TCT CGC CAG ACT CTC ACC TGG ATC CCA GAC TCC TTC TTC TCC AGT CTT 192
33 Ser Arg Gln Thr Leu Thr Trp Ile Pro Asp Ser Phe Phe Ser Ser Leu 48
193 CTG AGC GGA CGC ATC TCG ACG CTG AAA GAT GAG ACC GGA GCA ATC TTC 240
49 Leu Ser Gly Arg Ile Ser Thr Leu Lys Asp Glu Thr Gly Ala Ile Phe 64
241 ATC GAC AGG GAC CCT ACA GTC TTC GCC CCC ATC CTC AAC TTC CTG CGC 288
65 Ile Asp Arg Asp Pro Thr Val Phe Ala Pro Ile Leu Ash Phe Leu Arg 80
289 ACC AAA GAG TTG GAT CCC AGG GGT GTC CAC GGT TCC AGC CTC CTC CAT 336
81 Thr Lys Glu Leu Asp Pro Arg Gly Val His Gly Ser Ser Leu Leu His 96
337 GAA GCC CAG TTC TAT GGG CTC ACT CCT CTG GTT CGT CGC CTG CAG CTT 384
97 Glu Ala Gln Phe Tyr Gly Leu Thr Pro Leu Val Arg Arg Leu Gln Leu 112
385 CGA GAG GAG TTG GAT CGA TCT TCT TGT GGA AAC GTC CTC TTC AAT GGT 432
113 Arg Glu Glu Leu Asp Arg Ser Ser Cys Gly Asn Val Leu Phe Asn Gly 128
433 TAC CTG CCG CCA CCA GTG TTC CCA GTG AAG CGG CGG AAC CGG CAC AGC 480
129 Tyr Leu Pro Pro Pro Val Phe Pro Val Lys Arg Arg Asn Arg His Ser 144
481 CTA GTG GGG CCT CAG CAG CTA GGA GGA CGG CCA GCC CCT GTC CGA CGG 528
145 Leu Val Gly Pro Gln Gln Leu Gly Gly Arg Pro Ala Pro Val Arg Arg 160
529 AGC AAC ACG ATG CCC CCC AAC CTT GGC AAT GCA GGG CTG CTG GGC CGA 576
161 Ser Asn Thr Met Pro Pro Asn Leu Gly Asn Ala Gly Leu Leu Gly Arg 176
577 ATG CTG GAT GAG AAA ACC CCT CCC TCA CCC TCA GGA CAA CCT GAG GAG 624
177 Met Leu Asp Glu Lys Thr Pro Pro Ser Pro Ser Gly Gln Pro Glu Glu 192
625 CCG GGG ATG GTG CGC CTG GTG TGT GGA CAC CAT AAT TGG ATC GCT GTG 672
193 Pro Gly Met Val Arg Leu Val Cys Gly His His Asn Trp Ile Ala Val 208
673 GCC TAT ACC CAG TTT CTA GTC TGC TAC AGG TTG AAG GAA GCC TCT GGC 720
209 Ala Tyr Thr Gln Phe Leu Val Cys Tyr Arg Leu Lys Glu Ala Ser Gly 224
721 TGG CAG CTG GTG TTT TCC AGC CCC CGC CTG GAC TGG CCC ATC GAA CGA 768
225 Trp Gln Leu Val Phe Ser Ser Pro Arg Leu Asp Trp Pro Ile Glu Arg 240
769 CTG GCG CTC ACA GCC CGG GTG CAT GGT GGG GCT TTG GGT GAA CAT GAC 816
241 Leu Ala Leu Thr Ala Arg Val His Gly Gly Ala Leu Gly Glu His Asp 256
817 AAG ATG GTG GCA GCA GCC ACC GGC AGC GAG ATC CTG CTA TGG GCT CTG 864
257 Lys Met Val Ala Ala Ala Thr Gly Ser Glu Ile Leu Leu Trp Ala Leu 272
865 CAG GCG GAA GGC GGT GGC TCC GAG ATA GGG GTC TTT CAT CTG GGG GTG 912
273 Gln Ala Glu Gly Gly Gly Ser Glu Ile Gly Val Phe His Leu Gly Val 288
913 CCT GTG GAG GCC TTG TTC TTC GTC GGG AAC CAG CTC ATT GCT ACA AGC 960
289 Pro Val Glu Ala Leu Phe Phe Val Gly Asn Gln Leu Ile Ala Thr Ser 304
961 CAC ACA GGG CGC ATC GGG GTG TGG AAT GCC GTC ACC AAG CAC TGG CAG 1008
305 His Thr Gly Arg Ile Gly Val Trp Asn Ala Val Thr Lys His Trp Gln 320
1009 GTC CAG GAG GTG CAG CCC ATC ACC AGT TAT GAC GCG GCA GGC TCC TTC 1056
321 Val Gln Glu Val Gln Pro Ile Thr Ser Tyr Asp Ala Ala Gly Ser Phe 336
1057 CTC CTC CTG GGC TGC AAC AAC GGC TCC ATT TAC TAC GTG GAT GTG CAG 1104
337 Leu Leu Leu Gly Cys Asn Asn Gly Ser Ile Tyr Tyr Val Asp Val Gln 352
1105 AAG TTC CCC TTG CGC ATG AAA GAC AAC GAC CTC CTT GTC AGC GAG CTC 1152
353 Lys Phe Pro Leu Arg Met Lys Asp ASn Asp Leu Leu Val Ser Glu Leu 368
1153 TAT CGG GAC CCA GCG GAG GAT GGG GTC ACC GCC CTC AGT GTC TAC CTC 1200
369 Tyr Arg Asp Pro Ala Glu Asp Gly Val Thr Ala Leu Ser Val Tyr Leu 384
1201 ACC CCC AAG ACC AGT GAC AGT GGG AAC TGG ATC GAG ATC GCC TAT GGC 1248
385 Thr Pro Lys Thr Ser Asp Ser Gly Asn Trp Ile Glu Ile Ala Tyr Gly 400
1249 ACC AGC TCA GGG GGC GTG CGG GTC ATC GTG CAG CAC CCG GAG ACT GTG 1296
401 Thr Ser Ser Gly Gly Val Arg Val Ile Val Gln His Pro Glu Thr Val 416
1297 GGC TCG GGG CCT CAG CTC TTC CAG ACC TTC ACT GTG CAC CGC AGC CCT 1344
417 Gly Ser Gly Pro Gln Leu Phe Gln Thr Phe Thr Val His Arg Ser Pro 432
1345 GTC ACC AAG ATC ATG CTG TCG GAG AAG CAC CTC ATC TCA GTC TGT GCC 1392
433 Val Thr Lys Ile Met Leu Ser Glu Lys His Leu Ile Ser Val Cys Ala 448
1393 GAC AAC AAC CAC GTG CGG ACA TGG TCT GTG ACT CGC TTC CGC GGC ATG 1440
449 Asp Asn Asn His Val Arg Thr Trp Ser Val Thr Arg Phe Arg Gly Met 464
1441 ATT TCC ACC CAG CCC GGC TCC ACC CCA CTC GCT TCC TTT AAG ATC CTG 1488
465 Ile Ser Thr Gln Pro Gly Ser Thr Pro Leu Ala Ser Phe Lys Ile Leu 480
1489 GCT CTG GAG TCG GCA GAT GGG CAT GGC GGC TGC AGT GCT GGC AAT GAC 1536
481 Ala Leu Glu Ser Ala Asp Gly His Gly Gly Cys Ser Ala Gly Asn Asp 496
1537 ATT GGC CCC TAC GGT GAG CGG GAC GAC CAG CAA GTG TTC ATC CAG AAG 1584
497 Ile Gly Pro Tyr Gly Glu Arg Asp Asp Gln Gln Val Phe Ile Gln Lys 512
1585 GTG GTG CCC AGT GCC AGC CAG CTC TTC GTG CGT CTC TCA TCT ACT GGG 1632
513 Val Val Pro Ser Ala Ser Gln Leu Phe Val Arg Leu Ser Ser Thr Gly 528
1633 CAG CGG GTG TGC TCC GTG CGC TCC GTG GAC GGC TCA CCC ACG ACG GCC 1680
529 Gln Arg Val Cys Ser Val Arg Ser Val Asp Gly Ser Pro Thr Thr Ala 544
1681 TTC ACA GTG CTG GAG TGC GAG GGC TCC CGG CGG CTC GGC TCT CGG CCC 1728
545 Phe Thr Val Leu Glu Cys Glu Gly Ser Arg Arg Leu Gly Ser Arg Pro 560
1729 CGG CGC TAC CTG CTC ACT GGC CAG GCC AAC GGC AGC TTG GCC ATG TGG 1776
561 Arg Arg Tyr Leu Leu Thr Gly Gln Ala Asn Gly Ser Leu Ala Met Trp 576
1777 GAC CTA ACC ACC GCC ATG GAC GGC CTC GGC CAG GCC CCT GCA GGT GGC 1824
577 Asp Leu Thr Thr Ala Met Asp Gly Leu Gly Gln Ala Pro Ala Gly Gly 592
1825 CTG ACG GAG CAA GAG CTG ATG GAA CAG CTG GAA CAC TGT GAG CTG GCC 1872
593 Leu Thr Glu Gln Glu Leu Met Glu Gln Leu Glu His Cys Glu Leu Ala 608
1873 CCG CCG GCT CCT TCA GCT CCC TCA TGG GGC TGT CTC CCC AGC CCC TCA 1920
609 Pro Pro Ala Pro Ser Ala Pro Ser Trp Gly Cys Leu Pro Ser Pro Ser 624
1921 CCC CGC ATC TCC CTC ACC AGC CTC CAC TCA GCC TCC AGC AAC ACC TCC 1968
625 Pro Arg Ile Ser Leu Thr Ser Leu His Ser Ala Ser Ser Asn Thr Ser 640
1969 TTG TCT GGC CAC CGT GGG AGC CCA AGC CCC CCG CAG GCT GAG GCC CGG 2016
641 Leu Ser Gly His Arg Gly Ser Pro Ser Pro Pro G1n Ala Glu Ala Arg 656
2017 CGC CGT GGT GGG GGC AGC TTT GTG GAA CGC TGC CAG GAA CTG GTG CGG 2064
657 Arg Arg Gly Gly Gly Ser Phe Val Glu Arg Cys Gln Glu Leu Val Arg 672
2065 AGT GGG CCA GAC CTC CGA CGG CCA CCC ACA CCA GCC CCG TGG CCC TCC 2112
673 Ser Gly Pro Asp Leu Arg Arg Pro Pro Thr Pro Ala Pro Trp Pro Ser 688
2113 AGC GGT CTC GGC ACT CCC CTC ACA CCT CCC AAG ATG AAG CTC AAT GAA 2160
689 Ser Gly Leu Gly Thr Pro Leu Thr Pro Pro Lys Met Lys Leu ASn Glu 704
2161 ACT TCC TTT TGA ACA ACG CAG CTG CCA TGA TGC CTT GGG ATG CCC TGG 2208
705 Thr Ser Phe *** 708
2209 TCC TGG GGG ACT CAG GTG CCT CCC TGA TTC CTG TGG GAA CCC CGG GTT 2256
2257 CAG GGC CAG GGC CTC CTT GGA ATA AAT GGT TAT TGT TAC TAG GTC CCC 2304
2305 ACC TTC CCT CTT TTC TGG AAG CCA AAG TCA GCC TCC CCA ATA AAG TCC 2352
2353 TCA CTG CCA AAA AAA AAA AAA AAA AAA 2379
...
3．PP238
A: the nucleotide sequence (SEQ ID NO: 7) Length: 2023
GGGCTAGGGC CGGGGCCTGG CTGCGCGGCT GGGCCAAGGC CCGCGATGGT GATCTGCTGT 60
GCGGCCGTGA ACTGCTCCAA CCGGCAGGGA AAGGGCGAGA AGCGCGCCGT CTCCTTCCAC 120
AGGTTCCCCC TAAAGGACTC AAAACGTCTA ATCCAATGGT TAAAAGCTGT TCAGAGGGAT 180
AACTGGACTC CCACTAAGTA TTCATTTCTC TGTAGTGAGC ATTTCACCAA AGACAGCTTC 240
TCCAAGAGGC TGGAGGACCA GCATCGCCTG CTGAAGCCCA CGGCCGTGCC ATCCATCTTC 300
CACCTGACCG AGAAGAAGAG GGGGGCTGGA GGCCATGGCC GCACCCGGAG AAAAGATGCC 360
AGCAAGGCCA CAGGGGGTGT GAGGGGACAC TCGAGTGCCG CCACCGGCAG AGGAGCTGCA 420
GGTTGGTCAC CGTCCTCGAG TGGAAACCCG ATGGCCAAGC CAGAGTCCCG CAGGTTGAAG 480
CAAGCTGCTC TGCAAGGTGA AGCCACACCC AGGGCGGCCC AGGAGGCCGC CAGCCAGGAG 540
CAGGCCCAGC AAGCTCTGGA ACGGACTCCA GGAGATGGAC TGGCCACCAT GGTGGCAGGC 600
AGTCAGGGAA AAGCAGAAGC GTCTGCCACA GATGCTGGCG ATGAGAGCGC CACTTCCTCC 660
ATCGAAGGGG GCGTGACAGA TAAGAGTGGC ATTTCTATGG ATGACTTTAC GCCCCCAGGA 720
TCTGGGGCGT GCAAATTTAT CGGCTCACTT CATTCGTACA GTTTCTCCTC TAAGCACACC 780
CGAGAAAGGC CATCTGTCCC CCGAGAGCCC ATTGACCGCA AGAGGCTGAA GAAAGATGTG 840
GAACCAAGCT GCAGTGGGAG CAGCCTGGGA CCCGACAAGG GCCTGGCCCA GAGCCCTCCC 900
AGCTCATCAC TTACCGCGAC ACCGCAGAAG CCTTCCCAGA GCCCCTCTGC CCCTCCTGCC 960
GACGTCACCC CAAAGCCAGC CACGGAAGCC GTGCAGAGCG AGCACAGCGA CGCCAGCCCC 1020
ATGTCCATCA ACGAGGTCAT CCTGTCGGCG TCAGGGGCCT GCAAGCTCAT CGACTCACTG 1080
CACTCCTACT GCTTCTCCTC CCGGCAGAAC AAGAGCCAGG TGTGCTGCCT GCGGGAGCAG 1140
GTGGAGAAGA AGAACGGCGA GCTGAAGAGC CTGCGGCAGA GGGTCAGCCG CTCCGACAGC 1200
CAGGTGCGGA AGCTACAGGA GAAGCTGGAT GAGCTGAGGA GAGTGAGCGT CCCCTATCCA 1260
AGTAGCCTGC TGTCGCCCAG CCGCGAGCCC CCCAAGATGA ACCCAGTGGT GGAGCCACTG 1320
TCCTGGATGC TGGGCACCTG GCTGTCGGAC CCACCTGGAG CCGGGACCTA CCCCACACTG 1380
CAGCCCTTCC AGTACCTGGA GGAGGTTCAC ATCTCCCACG TGGGCCAGCC CATGCTGAAC 1440
TTCTCGTTCA ACTCCTTCCA CCCGGACACG CGCAAGCCGA TGCACAGAGA GTGTGGCTTC 1500
ATTCGCCTCA AGCCCGACAC CAACAAGGTG GCCTTTGTCA GCGCCCAGAA CACAGGCGTG 1560
GTGGAAGTGG AGGAGGGCGA GGTGAACGGG CAGGAGCTGT GCATCGCATC CCACTCCATC 1620
GCCAGGATCT CCTTCGCCAA GGAGCCCCAC GTAGAGCAGA TCACCCGGAA GTTCAGGCTG 1680
AATTCTGAAG GCAAACTTGA GCAGACGGTC TCCATGGCAA CCACGACACA GCCAATGACT 1740
CAGCATCTTC ACGTCACCTA CAAGAAGGTG ACCCCGTAAA CCTAGAGCTT CTGGAGCCCT 1800
CGGGAGGGCC TGGCTACTGT GCCTCAACGG TTCGGCTCCT CAACAGACAG TCCCTGCGGC 1860
AAAAGTGGGT GTGGCCGTGA GCCTCTGCAG GCTCAAGAGT GTTGTCCAGA TGTTTCTGTA 1920
CTGGCATAGA AAAACCAAAT AAAAGGCCTT TATTTTTATG GCTGAGGATT TTGAATATTA 1980
AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAA 2023
B: the amino acid sequence (SEQ ID NO: 8) Length: 577
1 MVICCAAVNC SNRQGKGEKR AVSFHRFPLK DSKRLIQWLK AVQRDNWTPT
51 KYSFLCSEHF TKDSFSKRLE DQHRLLKPTA VPSIFHLTEK KRGAGGHGRT
101 RRKDASKATG GVRGHSSAAT GRGAAGWSPS SSGNPMAKPE SRRLKQAALQ
151 GEATPRAAQE AASQEQAQQA LERTPGDGLA TMVAGSQGKA EASATDAGDE
201 SATSSIEGGV TDKSGISMDD FTPPGSGACK FIGSLHSYSF SSKHTRERPS
251 VPREPIDRKR LKKDVEPSCS GSSLGPDKGL AQSPPSSSLT ATPQKPSQSP
301 SAPPADVTPK PATEAVQSEH SDASPMSINE VILSASGACK LIDSLHSYCF
351 SSRQNKSQVC CLREQVEKKN GELKSLRQRV SRSDSQVRKL QEKLDELRRV
401 SVPYPSSLLS PSREPPKMNP VVEPLSWMLG TWLSDPPGAG TYPTLQPFQY
451 LEEVHISHVG QPMLNFSFNS FHPDTRKPMH RECGFIRLKP DTNKVAFVSA
501 QNTGVVEVEE GEVNGQELCI ASHSIARISF AKEPHVEQIT RKFRLNSEGK
551 LEQTVSMATT TQPMTQHLHV TYKKVTP
Clone: PP238 (SEQ ID NO: 9)
Start codon: 46 ATG termination codon: 1779TAA Protein Weight: 62856.51
1 GGG CTA GGG CCG GGG CCT GGC TGC GCG GCT GGG CCA AGG CCC GCG ATG 48
1 Met 1
49 GTG ATC TGC TGT GCG GCC GTG AAC TGC TCC AAC CGG CAG GGA AAG GGC 96
2 Val Ile Cys Cys Ala Ala Val Asn Cys Ser Asn Arg Gln Gly Lys Gly 17
97 GAG AAG CGC GCC GTC TCC TTC CAC AGG TTC CCC CTA AAG GAC TCA AAA 144
18 Glu Lys Arg Ala Val Ser Phe His Arg Phe Pro Leu Lys Asp Ser Lys 33
145 CGT CTA ATC CAA TGG TTA AAA GCT GTT CAG AGG GAT AAC TGG ACT CCC 192
34 Arg Leu Ile Gln Trp Leu Lys Ala Val Gln Arg Asp Asn Trp Thr Pro 49
193 ACT AAG TAT TCA TTT CTC TGT AGT GAG CAT TTC ACC AAA GAC AGC TTC 240
50 Thr Lys Tyr Ser Phe Leu Cys Ser Glu His Phe Thr Lys Asp Ser Phe 65
241 TCC AAG AGG CTG GAG GAC CAG CAT CGC CTG CTG AAG CCC ACG GCC GTG 288
66 Ser Lys Arg Leu Glu Asp Gln His Arg Leu Leu Lys Pro Thr Ala Val 81
289 CCA TCC ATC TTC CAC CTG ACC GAG AAG AAG AGG GGG GCT GGA GGC CAT 336
82 Pro Ser Ile Phe His Leu Thr Glu Lys Lys Arg Gly Ala Gly Gly His 97
337 GGC CGC ACC CGG AGA AAA GAT GCC AGC AAG GCC ACA GGG GGT GTG AGG 384
98 Gly Arg Thr Arg Arg Lys Asp Ala Ser Lys Ala Thr Gly Gly Val Arg 113
385 GGA CAC TCG AGT GCC GCC ACC GGC AGA GGA GCT GCA GGT TGG TCA CCG 432
114 Gly His Ser Ser Ala Ala Thr Gly Arg Gly Ala Ala Gly Trp Ser Pro 129
433 TCC TCG AGT GGA AAC CCG ATG GCC AAG CCA GAG TCC CGC AGG TTG AAG 480
130 Ser Ser Ser Gly Asn Pro Met Ala Lys Pro Glu Ser Arg Arg Leu Lys 146
481 CAA GCT GCT CTG CAA GGT GAA GCC ACA CCC AGG GCG GCC CAG GAG GCC 528
146 Gln Ala Ala Leu Gln Gly Glu Ala Thr Pro Arg Ala Ala Gln Glu Ala 161
529 GCC AGC CAG GAG CAG GCC CAG CAA GCT CTG GAA CGG ACT CCA GGA GAT 576
162 Ala Ser Gln Glu Gln Ala Gln Gln Ala Leu Glu Arg Thr Pro Gly Asp 177
577 GGA CTG GCC ACC ATG GTG GCA GGC AGT CAG GGA AAA GCA GAA GCG TCT 624
178 Gly Leu Ala Thr Met Val Ala Gly Ser Gln Gly Lys Ala Glu Ala Ser 193
625 GCC ACA GAT GCT GGC GAT GAG AGC GCC ACT TCC TCC ATC GAA GGG GGC 672
194 Ala Thr Asp Ala Gly Asp Glu Ser Ala Thr Ser Ser Ile Glu Gly Gly 209
673 GTG ACA GAT AAG AGT GGC ATT TCT ATG GAT GAC TTT ACG CCC CCA GGA 720
210 Val Thr Asp Lys Ser Gly Ile Ser Met Asp Asp Phe Thr Pro Pro Gly 225
721 TCT GGG GCG TGC AAA TTT ATC GGC TCA CTT CAT TCG TAC AGT TTC TCC 768
226 Ser Gly Ala Cys Lys Phe Ile Gly Ser Leu His Ser Tyr Ser Phe Ser 241
769 TCT AAG CAC ACC CGA GAA AGG CCA TCT GTC CCC CGA GAG CCC ATT GAC 816
242 Ser Lys His Thr Arg Glu Arg Pro Ser Val Pro Arg Glu Pro Ile Asp 257
817 CGC AAG AGG CTG AAG AAA GAT GTG GAA CCA AGC TGC AGT GGG AGC AGC 864
258 Arg Lys Arg Leu Lys Lys Asp Val Glu Pro Ser Cys Ser Gly Ser Ser 273
865 CTG GGA CCC GAC AAG GGC CTG GCC CAG AGC CCT CCC AGC TCA TCA CTT 912
274 Leu Gly Pro Asp Lys Gly Leu Ala Gln Ser Pro Pro Ser Ser Ser Leu 289
913 ACC GCG ACA CCG CAG AAG CCT TCC CAG AGC CCC TCT GCC CCT CCT GCC 960
290 Thr Ala Thr Pro Gln Lys Pro Ser Gln Ser Pro Ser Ala Pro Pro Ala 305
961 GAC GTC ACC CCA AAG CCA GCC ACG GAA GCC GTG CAG AGC GAG CAC AGC 1008
306 Asp Val Thr Pro Lys Pro Ala Thr Glu Ala Val Gln Ser Glu His Ser 321
1009 GAC GCC AGC CCC ATG TCC ATC AAC GAG GTC ATC CTG TCG GCG TCA GGG 1056
322 Asp Ala Ser Pro Met Ser Ile Asn Glu Val Ile Leu Ser Ala Ser Gly 337
1057 GCC TGC AAG CTC ATC GAC TCA CTG CAC TCC TAC TGC TTC TCC TCC CGG 1104
338 Ala Cys Lys Leu Ile Asp Ser Leu His Ser Tyr Cys Phe Ser Ser Arg 353
1105 CAG AAC AAG AGC CAG GTG TGC TGC CTG CGG GAG CAG GTG GAG AAG AAG 1152
354 Gln Asn Lys Ser Gln Val Cys Cys Leu Arg Glu Gln Val Glu Lys Lys 369
1153 AAC GGC GAG CTG AAG AGC CTG CGG CAG AGG GTC AGC CGC TCC GAC AGC 1200
370 Asn Gly Glu Leu Lys Ser Leu Arg Gln Arg Val Ser Arg Ser Asp Ser 385
1201 CAG GTG CGG AAG CTA CAG GAG AAG CTG GAT GAG CTG AGG AGA GTG AGC 1248
386 Gln Val Arg Lys Leu Gln Glu Lys Leu Asp Glu Leu Arg Arg Val Ser 401
1249 GTC CCC TAT CCA AGT AGC CTG CTG TCG CCC AGC CGC GAG CCC CCC AAG 1296
402 Val Pro Tyr Pro Ser Ser Leu Leu Ser Pro Ser Arg Glu Pro Pro Lys 417
1297 ATG AAC CCA GTG GTG GAG CCA CTG TCC TGG ATG CTG GGC ACC TGG CTG 1344
418 Met Asn Pro Val Val Glu Pro Leu Ser Trp Met Leu Gly Thr Trp Leu 433
1345 TCG GAC CCA CCT GGA GCC GGG ACC TAC CCC ACA CTG CAG CCC TTC CAG 1392
434 Ser Asp Pro Pro Gly Ala Gly Thr Tyr Pro Thr Leu Gln Pro Phe Gln 449
1393 TAC CTG GAG GAG GTT CAC ATC TCC CAC GTG GGC CAG CCC ATG CTG AAC 1440
450 Tyr Leu Glu Glu Val His Ile Ser His Val Gly Gln Pro Met Leu Asn 465
1441 TTC TCG TTC AAC TCC TTC CAC CCG GAC ACG CGC AAG CCG ATG CAC AGA 1488
466 Phe Ser Phe Asn Ser Phe His Pro Asp Thr Arg Lys Pro Met His Arg 481
1489 GAG TGT GGC TTC ATT CGC CTC AAG CCC GAC ACC AAC AAG GTG GCC TTT 1536
482 Glu Cys Gly Phe Ile Arg Leu Lys Pro Asp Thr Asn Lys Val Ala Phe 497
1537 GTC AGC GCC CAG AAC ACA GGC GTG GTG GAA GTG GAG GAG GGC GAG GTG 1584
498 Val Ser Ala Gln Asn Thr Gly Val Val Glu Val Glu Glu Gly Glu Val 513
1585 AAC GGG CAG GAG CTG TGC ATC GCA TCC CAC TCC ATC GCC AGG ATC TCC 1632
514 Asn Gly Gln Glu Leu Cys Ile Ala Ser His Ser Ile Ala Arg Ile Ser 529
1633 TTC GCC AAG GAG CCC CAC GTA GAG CAG ATC ACC CGG AAG TTC AGG CTG 1680
530 Phe Ala Lys Glu Pro His Val Glu Gln Ile Thr Arg Lys Phe Arg Leu 545
1681 AAT TCT GAA GGC AAA CTT GAG CAG ACG GTC TCC ATG GCA ACC ACG ACA 1728
546 Asn Ser Glu Gly Lys Leu Glu Gln Thr Val Ser Met Ala Thr Thr Thr 561
1729 CAG CCA ATG ACT CAG CAT CTT CAC GTC ACC TAC AAG AAG GTG ACC CCG 1776
562 Gln Pro Met Thr Gln His Leu His Val Thr Tyr Lys Lys Val Thr Pro 577
1777 TAA ACC TAG AGC TTC TGG AGC CCT CGG GAG GGC CTG GCT ACT GTG CCT 1824
578 *** 578
1825 CAA CGG TTC GGC TCC TCA ACA GAC AGT CCC TGC GGC AAA AGT GGG TGT 1872
1873 GGC CGT GAG CCT CTG CAG GCT CAA GAG TGT TGT CCA GAT GTT TCT GTA 1920
1921 CTG GCA TAG AAA AAC CAA ATA AAA GGC CTT TAT TTT TAT GGC TGA GGA 1968
1969 TTT TGA ATA TTA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA 2016
2017 AAA AAA A 2023
4. PP856
A: the nucleotide sequence (SEQ ID NO: 10) Length: 2364
GGAACACGTG CTTTCTGGGC AGGTCGCCCC TCAGTCTCCA CTAGAGACAG GACTGACCAG 60
TTGCTCTTCC TTCCAAGAAC CTTCGAGATC TGCGGTCTGG GGTCTGGTTG AAAGATGGCG 120
GCCCTCACTA CCCTGTTTAA GTACATAGAT GAAAATCAGG ATCGCTACAT TAAGAAACTC 180
GCAAAATGGG TGGCTATCCA GAGTGTGTCT GCGTGGCCGG AGAAGAGAGG CGAAATCAGG 240
AGGATGATGG AAGTTGCTGC TGCAGATGTT AAGCAGTTGG GGGGCTCTGT GGAACTGGTG 300
GATATCGGAA AACAAAAGGA GATTCCTGTC AACGTCCGAT TCTGCCTCGA AGGCATGGAG 360
GAGTCAGGCT CTGAGGGCCT AGACGAGCTG ATTTTTGCCC GGAAAGACAC ATTCTTTAAG 420
GATGTGGACT ATGTCTGCAT TTCTGACAAT TACTGGCTGG GAAAGAAGAA GCCCTGCATC 480
ACCTACGGCC TCAGGGGCAT TTGCTACTTT TTCATCGAGG TGGAGTGCAG CAACAAAGAC 540
CTCCATTCTG GGGTGTACGG GGGCTCGGTG CATGAGGCCA TGACTGATCT CATTTTGCTG 600
ATGGGCTCTT TGGTGGACAA GAGGGGGAAC ATCCTGATCC CCGGCATTAA CGAGGCCGTG 660
GCCGCCGTCA CGGAAGAGGA GCACAAGCTG TACGACGACA TCGACTTTGA CATAGAGGAG 720
TTTGCCAAGG ATGTGGGGGC GCAGATCCTC CTGCACAGCC ACAAGAAAGA CATCCTCATG 780
CACCGATGGC GGTACCCGTC TCTGTCCCTC CATGGCATCG AAGGCGCCTT CTCTGGGTCT 840
GGGGCCAAGA CCGTGATTCC CAGGAAGGTG GTTGGCAAGT TCTCCATCAG GCTCGTGCCG 900
AACATGACTC CTGAAGTCGT CGGCGAGCAG GTCACAAGCT ACCTAACTAA GAAGTTTGCT 960
GAACTACGCA GCCCCAATGA GTTCAAGGTG TACATGGGCC ACGGTGGGAA GCCCTGGGTC 1020
TCCGACTTCA GTCACCCTCA TTACCTGGCT GGGAGAAGAG CCATGAAGAC AGTTTTTGGT 1080
GTTGAGCCAG ACTTGACCAG GGAAGGCGGC AGTATTCCCG TGACCTTGAC CTTTCAGGAG 1140
GCCACGGGCA AGAACGTCAT GCTGCTGCCT GTGGGGTCAG CGGATGACGG AGCCCACTCC 1200
CAGAATGAAA AGCTCAACAG GTATAACTAC ATAGAGGGAA CCAAGATGCT GGCCGCGTAC 1260
CTGTATGAGG TCTCCCAGCT GAAGGACTAG GCCAAGCCCT CTGTGTGCCA TCTCCAATGA 1320
GAAGGAATCC TGCCCTCACC TCACCCTTTT CCAACTTGCC CAGGGAAGTG GAGGTTCCCT 1380
CTTTCCTTTC CCTCTTGTCA GGTCATCCAT GACTTTAGAG AACAGACACA AGTGTATCCA 1440
GCTGTCCACG GGTGGAGCTA CCCGTTGGGC TTATGAGTGA CCTGGAGTGA CAGCTGAGTC 1500
ACCCTGGGTA AGTTCTCAGA GTGGTCAGGA TGGCTTGACC TGCAGAAGAT ACCCAAGGTC 1560
CAAAAGCACA AGGTCTGCGG AAAGTTCTGG TTGTCGGCTG GGCACCACGG CTCACACCTA 1620
TAATCGAGCA CTTTGGGAGG CCAAGACAGG AGGATCACTT GAGGCCAGGA GTCTGAGACA 1680
AGCCTAGGCA ACAAAACAAG ACTCTGTCTC TACAAAAAGT TTAAGAAATG AGCCAGACAT 1740
GGTGGTGTAT GCCTGTAGTC CCAGCCACTC AGAAGGCTGA GGCAGGAGGA TCGCTTGAGA 1800
CCAAGAGTTT GAGCCTGCGG TGAGCTGTGA ATGCACCACG GCACTCAAGC CTGGGCAATG 1860
TAGCAAGATC CTGTCTCTAC AAGAAATTTT TTAAAAATGA GCCAAGTGTG GTGGTGCATG 1920
CCTGTAGTTC CAGCTACTCA GGACACTGAC GTAGGAGGGT TGCTTGAGAC TGAGAGTTGG 1980
AGGCTGCGAT GAGCCATGAA TGCCCCACTG CACTCCAGCC TGGGCGACAG AACGAGACCC 2040
CATCTCAAAA AAAATAAGTT CTGGTTGTCA TTGAATTGGG ATAAACAGAG AGCTTGATGC 2100
TTTCTGCCTT CTGTCTCAGG TGATGCATTG CACATTTGGG ATATTTGGAA AGGAAATGAG 2160
GAAAGAAATT AGGGCCTCCT CTGATCTCTC GCTATCTGCG GGTCCTGTCC TTTTCTCAAG 2220
ACCTTCACCA TTACTGGTGT TTTCCTGTCT TCTCTTTAGT ATGATCCCTC AAAACCTCAC 2280
TAACTGGAAG GATGATTTTG TCTCAGTTTG TACTCCTAAA TAAAAAGTAA ACATGACACC 2340
TCTAAAAAAA AAAAAAAAAA AAAA 2364
B: the amino acid sequence (SEQ ID NO: 11) Length: 391
1 MAALTTLFKY IDENQDRYIK KLAKWVAIQS VSAWPEKRGE IRRMMEVAAA
51 DVKQLGGSVE LVDIGKQKEI PVNVRFCLEG MEESGSEGLD ELIFARKDTF
101 FKDVDYVCIS DNYWLGKKKP CITYGLRGIC YFFIEVECSN KDLHSGVYGG
151 SVHEAMTDLI LLMGSLVDKR GNILIPGINE AVAAVTEEEH KLYDDIDFDI
201 EEFAKDVGAQ ILLHSHKKDI LMHRWRYPSL SLHGIEGAFS GSGAKTVIPR
251 KVVGKFSIRL VPNMTPEVVG EQVTSYLTKK FAELRSPNEF KVYMGHGGKP
301 WVSDFSHPHY LAGRRAMKTV PGVEPDLTRE GGSIPVTLTF QEATGKNVML
351 LPVGSADDGA HSQNEKLNRY NYIEGTKMLA AYLYEVSQLK D
Clone: PP856 (SEQ ID NO: 12)
Start codon: 115 ATG termination codon: 1290TAG Protein Weight: 43831.06
1 GGA ACA CGT GCT TTC TGG GCA GGT CGC CCC TCA GTC TCC ACT AGA GAC 48
49 AGG ACT GAC CAG TTG CTC TTC CTT CCA AGA ACC TTC GAG ATC TGC GGT 96
97 CTG GGG TCT GGT TGA AAG ATG GCG GCC CTC ACT ACC CTG TTT AAG TAC 144
1 Met Ala Ala Leu Thr Thr Leu Phe Lys Tyr 10
145 ATA GAT GAA AAT CAG GAT CGC TAC ATT AAG AAA CTC GCA AAA TGG GTG 192
11 Ile Asp Glu Asn Gln Asp Arg Tyr Ile Lys Lys Leu Ala Lys Trp Val 26
193 GCT ATC CAG AGT GTG TCT GCG TGG CCG GAG AAG AGA GGC GAA ATC AGG 240
27 Ala Ile Gln Ser Val Ser Ala Trp Pro Glu Lys Arg Gly Glu Ile Arg 42
241 AGG ATG ATG GAA GTT GCT GCT GCA GAT GTT AAG CAG TTG GGG GGC TCT 288
43 Arg Met Met Glu Val Ala Ala Ala Asp Val Lys Gln Leu Gly Gly Ser 58
289 GTG GAA CTG GTG GAT ATC GGA AAA CAA AAG GAG ATT CCT GTC AAC GTC 336
59 Val Glu Leu Val Asp Ile Gly Lys Gln Lys Glu Ile Pro Val Asn Val 74
337 CGA TTC TGC CTC GAA GGC ATG GAG GAG TCA GGC TCT GAG GGC CTA GAC 384
75 Arg Phe Cys Leu Glu Gly Met Qlu Glu Ser Gly Ser Glu Gly Leu Asp 90
385 GAG CTG ATT TTT GCC CGG AAA GAC ACA TTC TTT AAG GAT GTG GAC TAT 432
91 Glu Leu Ile Phe Ala Arg Lys Asp Thr Phe Phe Lys Asp Val Asp Tyr 106
433 GTC TGC ATT TCT GAC AAT TAC TGG CTG GGA AAG AAG AAG CCC TGC ATC 480
107 Val Cys Ile Ser Asp Asn Tyr Trp Leu Gly Lys Lys Lys Pro Cys Ile 122
481 ACC TAC GGC CTC AGG GGC ATT TGC TAC TTT TTC ATC GAG GTG GAG TGC 528
123 Thr Tyr Gly Leu Arg Gly Ile Cys Tyr Phe Phe Ile Glu Val Glu Cys 138
529 AGC AAC AAA GAC CTC CAT TCT GGG GTG TAC GGG GGC TCG GTG CAT GAG 576
139 Ser Asn Lys Asp Leu His Ser Gly Val Tyr Gly Gly Ser Val His Glu 154
577 GCC ATG ACT GAT CTC ATT TTG CTG ATG GGC TCT TTG GTG GAC AAG AGG 624
155 Ala Met Thr Asp Leu Ile Leu Leu Met Gly Ser Leu Val Asp Lys Arg 170
625 GGG AAC ATC CTG ATC CCC GGC ATT AAC GAG GCC GTG GCC GCC GTC ACG 672
171 Gly Asn Ile Leu Ile Pro Gly Ile Asn Glu Ala Val Ala Ala Val Thr 186
673 GAA GAG GAG CAC AAG CTG TAC GAC GAC ATC GAC TTT GAC ATA GAG GAG 720
187 Glu Glu Glu His Lys Leu Tyr Asp Asp Ile Asp Phe Asp Ile Glu Glu 202
721 TTT GCC AAG GAT GTG GGG GCG CAG ATC CTC CTG CAC AGC CAC AAG AAA 768
203 Phe Ala Lys Asp Val Gly Ala Gln Ile Leu Leu His Ser His Lys Lys 218
769 GAC ATC CTC ATG CAC CGA TGG CGG TAC CCG TCT CTG TCC CTC CAT GGC 816
219 Asp Ile Leu Met His Arg Trp Arg Tyr Pro Ser Leu Ser Leu His Gly 234
817 ATC GAA GGC GCC TTC TCT GGG TCT GGG GCC AAG ACC GTG ATT CCC AGG 864
235 Ile Glu Gly Ala Phe Ser Gly Ser Gly Ala Lys Thr Val Ile Pro Arg 250
865 AAG GTG GTT GGC AAG TTC TCC ATC AGG CTC GTG CCG AAC ATG ACT CCT 912
251 Lys Val Val Gly Lys Phe Ser Ile Arg Leu Val Pro Asn Met Thr Pro 266
913 GAA GTC GTC GGC GAG CAG GTC ACA AGC TAC CTA ACT AAG AAG TTT GCT 960
267 Glu Val Val Gly Glu Gln Val Thr Ser Tyr Leu Thr Lys Lys Phe Ala 282
961 GAA CTA CGC AGC CCC AAT GAG TTC AAG GTG TAC ATG GGC CAC GGT GGG 1008
283 Glu Leu Arg Ser Pro Asn Glu Phe Lys Val Tyr Met Gly His Gly Gly 298
1009 AAG CCC TGG GTC TCC GAC TTC AGT CAC CCT CAT TAC CTG GCT GGG AGA 1056
299 Lys Pro Trp Val Ser Asp Phe Ser His Pro His Tyr Leu Ala Gly Arg 314
1057 AGA GCC ATG AAG ACA GTT TTT GGT GTT GAG CCA GAC TTG ACC AGG GAA 1104
315 Arg Ala Met Lys Thr Val Phe Gly Val Glu Pro Asp Leu Thr Arg Glu 330
1105 GGC GGC AGT ATT CCC GTG ACC TTG ACC TTT CAG GAG GCC ACG GGC AAG 1152
331 Gly Gly Ser Ile Pro Val Thr Leu Thr Phe Gln Glu Ala Thr Gly Lys 346
1153 AAC GTC ATG CTG CTG CCT GTG GGG TCA GCG GAT GAC GGA GCC CAC TCC 1200
347 Asn Val Met Leu Leu Pro Val Gly Ser Ala Asp Asp Gly Ala His Ser 362
1201 CAG AAT GAA AAG CTC AAC AGG TAT AAC TAC ATA GAG GGA ACC AAG ATG 1248
363 Gln Asn Glu Lys Leu Asn Arg Tyr Asn Tyr Ile Glu Gly Thr Lys Met 378
1249 CTG GCC GCG TAC CTG TAT GAG GTC TCC CAG CTG AAG GAC TAG GCC AAG 1296
379 Leu Ala Ala Tyr Leu Tyr Glu Val Ser Gln Leu Lys Asp *** 392
1297 CCC TCT GTG TGC CAT CTC CAA TGA GAA GGA ATC CTG CCC TCA CCT CAC 1344
1345 CCT TTT CCA ACT TGC CCA GGG AAG TGG AGG TTC CCT CTT TCC TTT CCC 1392
1393 TCT TGT CAG GTC ATC CAT GAC TTT AGA GAA CAG ACA CAA GTG TAT CCA 1440
1441 GCT GTC CAC GGG TGG AGC TAC CCG TTG GGC TTA TGA GTG ACC TGG AGT 1488
1489 GAC AGC TGA GTC ACC CTG GGT AAG TTC TCA GAG TGG TCA GGA TGG CTT 1536
1537 GAC CTG CAG AAG ATA CCC AAG GTC CAA AAG CAC AAG GTC TGC GGA AAG 1584
1585 TTC TGG TTG TCG GCT GGG CAC CAC GGC TCA CAC CTA TAA TCG AGC ACT 1632
1633 TTG GGA GGC CAA GAC AGG AGG ATC ACT TGA GGC CAG GAG TCT GAG ACA 1680
1681 AGC CTA GGC AAC AAA ACA AGA CTC TGT CTC TAC AAA AAG TTT AAG AAA 1728
1729 TGA GCC AGA CAT GGT GGT GTA TGC CTG TAG TCC CAG CCA CTC AGA AGG 1776
1777 CTG AGG CAG GAG GAT CGC TTG AGA CCA AGA GTT TGA GCC TGC GGT GAG 1824
1825 CTG TGA ATG CAC CAC GGC ACT CAA GCC TGG GCA ATG TAG CAA GAT CCT 1872
1873 GTC TCT ACA AGA AAT TTT TTA AAA ATG AGC CAA GTG TGG TGG TGC ATG 1920
1921 CCT GTA GTT CCA GCT ACT CAG GAC ACT GAC GTA GGA GGG TTG CTT GAG 1968
1969 ACT GAG AGT TGG AGG CTG CGA TGA GCC ATG AAT GCC CCA CTG CAC TCC 2016
2017 AGC CTG GGC GAC AGA ACG AGA CCC CAT CTC AAA AAA AAT AAG TTC TGG 2064
2065 TTG TCA TTG AAT TGG GAT AAA CAG AGA GCT TGA TGC TTT CTG CCT TCT 2112
2113 GTC TCA GGT GAT GCA TTG CAC ATT TGG GAT ATT TGG AAA GGA AAT GAG 2160
2161 GAA AGA AAT TAG GGC CTC CTC TGA TCT CTC GCT ATC TGC GGG TCC TGT 2208
2209 CCT TTT CTC AAG ACC TTC ACC ATT ACT GGT GTT TTC CTG TCT TCT CTT 2256
2257 TAG TAT GAT CCC TCA AAA CCT CAC TAA CTG GAA GGA TGA TTT TGT CTC 2304
2305 AGT TTG TAC TCC TAA ATA AAA AGT AAA CAT GAC ACC TCT AAA AAA AAA 2352
2353 AAA AAA AAA AAA 2364
5. PP1065
A: the nucleotide sequence (SEQ ID NO: 13) Length: 1910
GGGAAGTAGA AGACAGCGGC GTTGCCATGG CGGCGTCTCT GGGGCAGGTG TTGGCTCTGG 60
TGCTGGTGGC CGCTCTGTGG GGTGGCACGC AGCCGCTGCT GAAGCGGGCC TCCGCCGGCC 120
TGCAGCGGGT TCATGAGCCG ACCTGGGCCC AGCAGTTGCT ACAGGAGATG AAGACCCTCT 180
TCTTGAATAC TGAGTACCTG ATGCCCTTTC TCCTCAACCA GTGTGGATCC CTTCTCTATT 240
ACCTCACCTT GGCATCGACA GATCTGACCC TGGCTGTGCC CATCTGTAAC TCTCTGGCTA 300
TCATCTTCAC ACTGATTGTT GGGAAGGCCC TTGGAGAAGA TATTGGTGGA AAACGAGCAG 360
TTGCTGGCAT GGTGCTCACC GTGATAGGAA TTTCACTCTG CATCACAAGC TCAGTTCCAT 420
GGACTGCAGA ACTCCAGCTG CATGGAAAGG GCCAGCTGCA GACTTTGAGC CAGAAATGCA 480
AACGGGAGGC CTCTGGGACT CAGTCAGAGC GCTTTGGCTG AATGAGGGGT GGAACCGAGG 540
GAAGAAGGTA GAGAGCTGTG AGCCCCAGCC CCACCTGACT CCAGCACACC TGGCGAGTAG 600
TAGCTGTCAA TAAATCTATG GTAAACAGAC AAGAGGAGGT GGAAGGCCAT ACAGAATGGA 660
GCCGTGAGTA TGGCCAGCCT CCAGCTCTCA GCCAGGAGGT CCCCAACCCC AAGGAAGGAA 720
GAAACTGGAA ATTAGGAACT GCTTCCTCAT TTAACAAGGT AGGAAGTTAG GAGATCATTT 780
ACTTTCAATC ACAAGGGAGG AGAACTGTTC CTGGGGCCCA AGGCTGCCAG TTTCCAGCTC 840
AGAGCTCCTC CCACCCCAAC ATACTGTTTC CTGATCCAAC AGCTTACCTG ATCCAAGGGC 900
TCCTCTGTCT GAGTGTCTTC ATCATCTGCT ACTTCCAGGG CCCCCGCTGT CTCCTTCCTT 960
CGGTGGGGAG CTCCATATAC TCCTATAACT CCTAAAGAGG GGAGGCAGCA CTGGGTGATG 1020
CCCAGGCTGA AGAGCCCTCA GCATTCCTGC CCTGGACCTT CATGCTAGCC CCTTTCCCTC 1080
CCCTGAACCT GGTTCTGCAT TCCCCACCAC CTCCCAGGAT GGCAAGGAAG TGAGAGGTGG 1140
GCCTTTGGTC CCACCCCCAT CCCCTCTATA TCCCACCCCT GAAGTCTTAT CGCTTTAAGC 1200
ACTGCCCTTT CCAGGTGCTT CTTTTCATGT GATGAGGCCC TGTGAAGAAG GGACAGGATA 1260
TACAGACGGG GGCAGCTGGA GACAGTTATG ATGAGTGCCG GCTTTGTGTC TGAGCATTCT 1320
GCTCCCATGG ACATCCCCAA CAACAGCAGG GACCAACCTA TGTCACTGTC AAAGGGCAGC 1380
TGAGAAGGGC CTGAGCCCCA GGGACCCCTC ACCTGATGGG AATGAGAGTG TGGGGAGCTT 1440
GCTTCTTGGC TGAATGGTCT GCTGGGGTCT GGCATAGAAA GCAGATGGCT TAATTCGGTC 1500
TGGTTCCTTT GGAGAGGGCT GGGTTACCTG GGCCCTGTGG CCTTGGGCCT AGAGAAGGGA 1560
CACTGGGCTT GGACCCTGAT TGCTGGCCAT TCTTACCTTT CCTACTCCCC AGTTCAGGCT 1620
TCAGAGAGCC CCTGACGCCT GCAGGAACAT GGCAGAGGAG ACACCGTCTG TCTTCACAAA 1680
GTACGTCCTC CCTCCTTGCT GCCTCTTCCC ACCAGCCTGA CTTATCCAGG GACAGAGCAG 1740
TAGATGCCTG GCACTCCTCG ATGCCCAGTG AAACCAGACT GTGCTTCCCC ACCCCCACCA 1800
CCATGCCCCA TGCTCACTGG CTCATTTCTT GGGAGGGCTT AGAGCTGGAT AATATAAGTT 1860
CCCTTGGGAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA 1910
B: the amino acid sequence (SEQ ID NO: 14) Length: 164
1 MAASLGQVLA LVLVAALWGG TQPLLKRASA GLQRVHEPTW AQQLLQEMKT
51 LFLNTEYLMP FLLNQCGSLL YYLTLASTDL TLAVPICNSL AIIFTLIVGK
101 ALGEDIGGKR AVAGMVLTVI GISLCITSSV PWTAELQLHG KGQLQTLSQK
151 CKREASGTQS ERFG
Clone: PP1065 (SEQ ID NO: 15)
Start codon: 27 ATG termination codon: 521TGA Protein Weight: 17599.78
1 GG GAA GTA GAA GAC AGC GGC GTT GCC ATG GCG GCG TCT CTG GGG CAG 47
1 Met Ala Ala Ser Leu Gly Gln 7
48 GTG TTG GCT CTG GTG CTG GTG GCC GCT CTG TGG GGT GGC ACG CAG CCG 95
8 Val Leu Ala Leu Val Leu Val Ala Ala Leu Trp Gly Gly Thr Gln Pro 23
96 CTG CTG AAG CGG GCC TCC GCC GGC CTG CAG CGG GTT CAT GAG CCG ACC 143
24 Leu Leu Lys Arg Ala Ser Ala Gly Leu Gln Arg Val His Glu Pro Thr 39
144 TGG GCC CAG CAG TTG CTA CAG GAG ATG AAG ACC CTC TTC TTG AAT ACT 191
40 Trp Ala Gln Gln Leu Leu Gln Glu Met Lys Thr Leu Phe Leu Asn Thr 55
192 GAG TAC CTG ATG CCC TTT CTC CTC AAC CAG TGT GGA TCC CTT CTC TAT 239
56 Glu Tyr Leu Met Pro Phe Leu Leu Asn Gln Cys Gly Ser Leu Leu Tyr 71
240 TAC CTC ACC TTG GCA TCG ACA GAT CTG ACC CTG GCT GTG CCC ATC TGT 287
72 Tyr Leu Thr Leu Ala Ser Thr Asp Leu Thr Leu Ala Val Pro Ile Cys 87
288 AAC TCT CTG GCT ATC ATC TTC ACA CTG ATT GTT GGG AAG GCC CTT GGA 335
88 Asn Ser Leu Ala Ile Ile Phe Thr Leu Ile Val Gly Lys Ala Leu Gly 103
336 GAA GAT ATT GGT GGA AAA CGA GCA GTT GCT GGC ATG GTG CTC ACC GTG 383
104 Glu Asp Ile Gly Gly Lys Arg Ala Val Ala Gly Met Val Leu Thr Val 119
384 ATA GGA ATT TCA CTC TGC ATC ACA AGC TCA GTT CCA TGG ACT GCA GAA 431
120 Ile Gly Ile Ser Leu Cys Ile Thr Ser Ser Val Pro Trp Thr Ala Glu 135
432 CTC CAG CTG CAT GGA AAG GGC CAG CTG CAG ACT TTG AGC CAG AAA TGC 479
136 Leu Gln Leu His Gly Lys Gly Gln Leu Gln Thr Leu Ser Gln Lys Cys 151
480 AAA CGG GAG GCC TCT GGG ACT CAG TCA GAG CGC TTT GGC TGA ATG AGG 527
152 Lys Arg Glu Ala Ser Gly Thr Gln Ser Glu Arg Phe Gly *** 165
528 GGT GGA ACC GAG GGA AGA AGG TAG AGA GCT GTG AGC CCC AGC CCC ACC 575
576 TGA CTC CAG CAC ACC TGG CGA GTA GTA GCT GTC AAT AAA TCT ATG GTA 623
624 AAC AGA CAA GAG GAG GTG GAA GGC CAT ACA GAA TGG AGC CGT GAG TAT 671
672 GGC CAG CCT CCA GCT CTC AGC CAG GAG GTC CCC AAC CCC AAG GAA GGA 719
720 AGA AAC TGG AAA TTA GGA ACT GCT TCC TCA TTT AAC AAG GTA GGA AGT 767
768 TAG GAG ATC ATT TAC TTT CAA TCA CAA GGG AGG AGA ACT GTT CCT GGG 815
816 GCC CAA GGC TGC CAG TTT CCA GCT CAG AGC TCC TCC CAC CCC AAC ATA 863
864 CTG TTT CCT GAT CCA ACA GCT TAC CTG ATC CAA GGG CTC CTC TGT CTG 911
912 AGT GTC TTC ATC ATC TGC TAC TTC CAG GGC CCC CGC TGT CTC CTT CCT 959
960 TCG GTG GGG AGC TCC ATA TAC TCC TAT AAC TCC TAA AGA GGG GAG GCA 1007
1008 GCA CTG GGT GAT GCC CAG GCT GAA GAG CCC TCA GCA TTC CTG CCC TGG 1055
1056 ACC TTC ATG CTA GCC CCT TTC CCT CCC CTG AAC CTG GTT CTG CAT TCC 1103
1104 CCA CCA CCT CCC AGG ATG GCA AGG AAG TGA GAG GTG GGC CTT TGG TCC 1151
1152 CAC CCC CAT CCC CTC TAT ATC CCA CCC CTG AAG TCT TAT CGC TTT AAG 1199
1200 CAC TGC CCT TTC CAG GTG CTT CTT TTC ATG TGA TGA GGC CCT GTG AAG 1247
1248 AAG GGA CAG GAT ATA CAG ACG GGG GCA GCT GGA GAC AGT TAT GAT GAG 1295
1296 TGC CGG CTT TGT GTC TGA GCA TTC TGC TCC CAT GGA CAT CCC CAA CAA 1343
1344 CAG CAG GGA CCA ACC TAT GTC ACT GTC AAA GGG CAG CTG AGA AGG GCC 1391
1392 TGA GCC CCA GGG ACC CCT CAC CTG ATG GGA ATG AGA GTG TGG GGA GCT 1439
1440 TGC TTC TTG GCT GAA TGG TCT GCT GGG GTC TGG CAT AGA AAG CAG ATG 1487
1488 GCT TAA TTC GGT CTG GTT CCT TTG GAG AGG GCT GGG TTA CCT GGG CCC 1535
1536 TGT GGC CTT GGG CCT AGA GAA GGG ACA CTG GGC TTG GAC CCT GAT TGC 1583
1584 TGG CCA TTC TTA CCT TTC CTA CTC CCC AGT TCA GGC TTC AGA GAG CCC 1631
1632 CTG ACG CCT GCA GGA ACA TGG CAG AGG AGA CAC CGT CTG TCT TCA CAA 1679
1680 AGT ACG TCC TCC CTC CTT GCT GCC TCT TCC CAC CAG CCT GAC TTA TCC 1727
1728 AGG GAC AGA GCA GTA GAT GCC TGG CAC TCC TCG ATG CCC AGT GAA ACC 1775
6.
7.
...
Claims (11)
1. isolating people's albumen with cancer suppressing function, it is characterized in that it comprises the polypeptide of the aminoacid sequence with the group of being selected from down: SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20;
Or its conservative property variation polypeptide or its active fragments or its reactive derivative.
2. polypeptide as claimed in claim 1, it is characterized in that this polypeptide is the polypeptide with aminoacid sequence of the group of being selected from down: SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQID NO:20.
3. isolating polynucleotide is characterized in that, 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:
(a) coding is as the polynucleotide of polypeptide as described in claim 1 and 2;
(b) with polynucleotide (a) complementary polynucleotide.
4. polynucleotide as claimed in claim 3, it is characterized in that the polypeptide of this polynucleotide encoding has the aminoacid sequence of the group of being selected from down: SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ IDNO:17, SEQ ID NO:20.
5. polynucleotide as claimed in claim 3 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:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21.
6. a carrier is characterized in that, it contains the described polynucleotide of claim 3.
7. 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 6;
(b) host cell that transforms or transduce with the described polynucleotide of claim 3.
8. the preparation method of the polypeptide of the people's protein-active with cancer suppressing function is characterized in that this method comprises:
(a) have under the proteic condition of people of cancer suppressing function suitable the expression, cultivate the described host cell of claim 7;
(b) from culture, isolate the polypeptide of people's protein-active with cancer suppressing function.
9. energy and the described people's protein-specific bonded antibody of claim 1 with cancer suppressing function.
10. nucleic acid molecule, it contains a successive 10-800 Nucleotide in the described polynucleotide of claim 3.
11. a pharmaceutical composition is characterized in that, it contains the described polypeptide of claim 1 and the pharmaceutically acceptable carrier of safe and effective amount.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011045450A CN1194989C (en) | 2000-02-17 | 2001-02-13 | Novel human protein able to suppress cancer cell growth and its coding sequence |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN00111697.5 | 2000-02-17 | ||
| CN00111697 | 2000-02-17 | ||
| CNB011045450A CN1194989C (en) | 2000-02-17 | 2001-02-13 | Novel human protein able to suppress cancer cell growth and its coding sequence |
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| Publication Number | Publication Date |
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| CN1309135A true CN1309135A (en) | 2001-08-22 |
| CN1194989C CN1194989C (en) | 2005-03-30 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB011045450A Expired - Fee Related CN1194989C (en) | 2000-02-17 | 2001-02-13 | Novel human protein able to suppress cancer cell growth and its coding sequence |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1671843B (en) * | 2002-07-25 | 2010-07-14 | 索尼株式会社 | Factor participating in transcriptional regulation |
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2001
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1671843B (en) * | 2002-07-25 | 2010-07-14 | 索尼株式会社 | Factor participating in transcriptional regulation |
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| Publication number | Publication date |
|---|---|
| CN1194989C (en) | 2005-03-30 |
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