CN1169955C - Human protein able to suppress growth of cancer cells and its coding sequence - Google Patents

Human protein able to suppress growth of cancer cells and its coding sequence Download PDF

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CN1169955C
CN1169955C CNB001119494A CN00111949A CN1169955C CN 1169955 C CN1169955 C CN 1169955C CN B001119494 A CNB001119494 A CN B001119494A CN 00111949 A CN00111949 A CN 00111949A CN 1169955 C CN1169955 C CN 1169955C
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CN1313298A (en
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顾健人
杨胜利
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Shanghai Cancer Institute
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Shanghai Cancer Institute
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Abstract

The present invention discloses a novel human protein with the function of inhibiting cancer, polynucleotide for encoding the polypeptide and a method for preparing the polypeptide by a recombinant technology. The present invention also discloses a method of using the polypeptide to treat various diseases, such as cancers. The present invention also discloses an antagonist of the polypeptide and a therapeutic effect thereof. The present invention also discloses the application of the polynucleotide for encoding the human protein with the function of inhibiting cancer.

Description

The proteic polynucleotide of people that coding has anticancer growth function
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 IDNO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ IDNO:26, SEQ ID NO:29, SEQ ID NO:32; 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 IDNO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ IDNO:20, SEQ ID NO:23, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:32.
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 IDNO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ IDNO:26, SEQ ID NO:29, SEQ ID NO:32.More preferably, the sequence of these polynucleotide is selected from down group: coding region sequence or the full length sequence of SEQ IDNO: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, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:33.
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.Be example with PP610 albumen (in this application, its clone numbering is adopted in proteinic name), the coding region sequence of encoding mature polypeptide can be identical with the coding region sequence shown in the SEQ ID NO: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.With PP784 albumen is example, and the coding region sequence of encoding mature polypeptide can be identical with the coding region sequence shown in the SEQID NO:6 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:5, but with the differentiated nucleotide sequence of coding region sequence shown in the SEQ ID NO:6.Have the albumen of cancer suppressing function for other, can the rest may be inferred.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, A Laboratory Manual, Cold Spring Harbor Laboratory.New York, 1989).Also can obtain the cDNA library of commercial offers, as the different cDNA library of Clontech company.When being used in combination the polymeric enzyme reaction technology, even few expression product also can be cloned.
Available ordinary method is screened gene of the present invention from these cDNA libraries.These methods include, but is not limited to: (1) DNA-DNA or DNA-RNA hybridization; (2) function of marker gene occurs or forfeiture; (3) mensuration has the level of the proteic transcript of cancer suppressing function; (4), detect the protein product of genetic expression by immunological technique or mensuration biologic activity.Aforesaid method can singly be used, but also several different methods combined utilization.
In (1) kind method, hybridizing used probe is and any a part of homology of polynucleotide of the present invention that at least 15 Nucleotide of its length better are at least 30 Nucleotide, are more preferably at least 50 Nucleotide, preferably at least 100 Nucleotide.In addition, the length of probe within 2kb, preferably is within the 1kb usually.Probe used herein is the dna sequence dna of chemosynthesis on the basis of gene DNA sequence information of the present invention normally.Gene of the present invention itself or fragment are certainly as probe.The mark of dna probe can be used radio isotope, fluorescein or enzyme (as alkaline phosphatase) etc.
In (4) kind method, detect the protein product of protein gene expression and can use immunological technique such as Western blotting, radioimmunoprecipitation, enzyme-linked immunosorbent assay (ELISA) etc. with cancer suppressing function.
Use method (Saiki, the et al.Science 1985 of round pcr DNA amplification/RNA; 230:1350-1354) be optimized for acquisition gene of the present invention.When particularly being difficult to obtain the cDNA of total length from the library, can preferably use RACE method (the terminal rapid amplifying method of RACE-cDNA), the primer that is used for PCR can suitably be selected according to sequence information of the present invention disclosed herein, and available ordinary method is synthetic.Available ordinary method is as the DNA/RNA fragment by gel electrophoresis separation and purifying amplification.
The gene of the present invention that obtains as mentioned above, perhaps the available ordinary method of mensuration of the nucleotide sequence of various dna fragmentations etc. such as dideoxy chain termination (Sanger et al.PNAS, 1977,74:5463-5467).This class nucleotide sequencing is available commercial sequencing kit etc. also.In order to obtain the cDNA sequence of total length, order-checking need be carried out repeatedly.Sometimes need to measure a plurality of clones' cDNA sequence, just can be spliced into the cDNA sequence of total length.
The present invention also relates to comprise the carrier of polynucleotide of the present invention, and the host cell that produces through genetically engineered with carrier of the present invention or albumen coded sequence with cancer suppressing function, and the method that produces polypeptide of the present invention through recombinant technology.
By the recombinant DNA technology of routine, can utilize polymerized nucleoside acid sequence of the present invention to can be used to express or produce the protein polypeptide with cancer suppressing function (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 SpringHarbor Laboratory.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 P LPromotor; Eukaryotic promoter comprises LTRs and some other known may command gene expression promoter in protokaryon or eukaryotic cell or its virus of CMV immediate early promoter, 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 above methods can wrap by in cell, extracellular or on cytolemma, express or be secreted into the extracellular.If desired, can utilize its physics, the separating by various separation methods with other characteristic and the albumen of purification of Recombinant of chemistry.These methods are well-known to those skilled in the art.The example of these methods includes, but are not limited to: conventional renaturation handles, with protein precipitant handle (salt analysis method), centrifugal, the broken bacterium of infiltration, superly handle, the combination of super centrifugal, sieve chromatography (gel-filtration), adsorption chromatography, ion exchange chromatography, high performance liquid chromatography (HPLC) and other various liquid chromatography (LC) technology and these methods.
The people's albumen or the polypeptide with cancer suppressing function of reorganization are of use in many ways.These purposes include, but is not limited to: directly have the disease due to the low or forfeiture of the protein function of cancer suppressing function as pharmacological agent and be used to screen and promote or antagonism has antibody, polypeptide or other part of the protein function of cancer suppressing function.For example, antibody can be used for activating or suppressing to have the proteic function of people of cancer suppressing function.The people's protein screening peptide library that has a cancer suppressing function with the reorganization of expressing can be used for seeking the peptide molecule that can suppress or stimulate the people's protein function with cancer suppressing function of therapeutic value.
The present invention also provides screening of medicaments to improve (agonist) or check the method that (antagonist) has the proteic medicament of people of cancer suppressing function to identify.Agonist improves the biological function such as stimulate cellular proliferation of the people's albumen with cancer suppressing function, and antagonist prevention disorder such as the various cancer relevant with cell hyperproliferation with treatment.For example, can be in the presence of medicine, the proteic film preparation of people that mammalian cell or expression is had cancer suppressing function is cultivated with the people's albumen with cancer suppressing function of mark.Measure the medicine raising then or check this interactional ability.
The proteic antagonist of people with cancer suppressing function comprises antibody, compound, acceptor disappearance thing and the analogue etc. that filter out.The proteic antagonist of people with cancer suppressing function can and be eliminated its function with the people's protein binding with cancer suppressing function, or suppresses to have the proteic generation of people of cancer suppressing function, or combines with the avtive spot of polypeptide and to make polypeptide can not bring into play biological function.The proteic antagonist of people with cancer suppressing function can be used for therepic use.
In screening during as the compound of antagonist, 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 be found in existing document (Sambrook, etal.).The people protein gene of reorganization with cancer suppressing function can be packaged in the liposome and be transferred in the cell in addition.
Suppress to have cancer suppressing function people's protein mRNA oligonucleotide (comprising sense-rna and DNA) and ribozyme also within the scope of the invention.Ribozyme is the enzyme sample RNA molecule that a kind of energy specificity is decomposed specific RNA, and its mechanism of action is to carry out the endonuclease effect after ribozyme molecule and the hybridization of complementary target RNA-specific.The RNA of antisense and DNA and ribozyme can obtain with existing any RNA or DNA synthetic technology, as the technology widespread use of solid phase phosphoamide chemical synthesis synthetic oligonucleotide.Antisense rna molecule can be transcribed acquisition by the dna sequence dna of this RNA that encodes in external or body.This dna sequence dna has been incorporated into the downstream of rna polymerase promoter of carrier.In order to increase the stability of nucleic acid molecule, available several different methods is modified it, and as increasing the sequence length of both sides, the connection between the ribonucleoside is used phosphoric acid thioester bond or peptide bond but not phosphodiester bond.
Polynucleotide imports tissue or intracellular method comprises: directly be injected into polynucleotide in the in-vivo tissue; Or external by carrier (as virus, phage or plasmid etc.) earlier with the polynucleotide transfered cell in, again cell is transplanted in the body etc.
Polypeptide of the present invention also can be used as the peptide spectrum analysis, for example, the polypeptide available physical, chemistry or enzyme carry out the specificity cutting, and carry out the two-dimentional or three-dimensional gel electrophoresis analysis of one dimension.
The present invention also provides the antibody at the people's proteantigen determinant with cancer suppressing function.These antibody include, but is not limited to: the fragment that polyclonal antibody, monoclonal antibody, chimeric antibody, single-chain antibody, Fab fragment and Fab expression library produce.
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.PatNo.4946778) also can be used for producing the anti-proteic single-chain antibody of people with cancer suppressing function.
Can be incorporated into the rondom polypeptide storehouse that solid formation forms by the various amino acid that may make up by screening with the protein bound peptide molecule of the 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.At present, need to identify the concrete site of each gene on the karyomit(e).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, Mendelian Inheritance in Man (can by with the online acquisition of Johns Hopkins University Welch Medical Library).Can pass through linkage analysis then, determine gene and navigated to relation between the disease on the chromosomal region already.
Then, need to measure ill and not cDNA between diseased individuals or genome sequence difference.If observe certain sudden change in some or all of diseased individuals, and this sudden change is not observed in any normal individual, then this sudden change may be the cause of disease of disease.More ill and diseased individuals not is usually directed at first seek the variation of structure in the karyomit(e), as from the horizontal visible of karyomit(e) or use based on detectable disappearance of the PCR of cDNA sequence or transposition.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 LaboratoryPress, 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
PP610, PP784, PP827, PP1057, PP1551, PP1553, PP1665, PP2135, PP2281, PP2447, PP3111 obtains by making up the human placenta cDNA library with ordinary method.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 XL 10-Gold recipient cell, obtained 1 * 10 6The cDNA library of cfu/ μ g cDNA titre.The first round is picking cDNA clone at random, is probe with high abundance cDNA clone with the cDNA clone who has proved cancer inhibitor cell growth function thereafter, screening by hybridization cDNA library, weak positive and negative clone of picking.With Qiagen 96 orifice plate plasmid extraction test kits, carry out the extraction of plasmid DNA by shop instruction.Plasmid DNA and empty carrier transfection simultaneously hepatoma cell line 7721.After the 100ng DNA alcohol precipitation drying, add 6 μ lH 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 that above clone has the cell clone of inhibition formation effect, the result is as shown in table 1 below.
Table 1cDNA clone's transfectional cell (7721) clone formation situation
CDNA clones title CDNA clones number (three repetitions) Empty carrier clone number (three repetitions)
PP610 0 0 0 21 30 42
PP784 1 3 2 18 24 25
PP827 5 3 1 31 38 32
PP1057 0 1 0 32 39 37
PP1551 0 0 0 27 25 26
PP1553 20 0 0 27 25 26
PP1665 17 18 18 22 25 13
PP2135 2 0 0 32 31 34
PP2281 0 0 0 41 36 33
PP2447 3 1 3 34 31 30
PP3111 0 1 0 12 18 20
Above-mentioned 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 (SEQ ID NO:1,4,7,10,13,16,19,22,25,28,31).
Embodiment 2: PCR obtains full-length gene from placenta cDNA:
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.With MMLV-RT-Superscript II (GIBCO BRL), ThermoScript II is carried out reverse transcription reaction at 42 ℃, obtains placenta cDNA.Utilize the special primer (as shown in the table) of each gene, by 97 ℃ of 3 ' 1 circulation.72 ℃ of 1 ' 35 circulation of 94C 30 " 60 ℃ 30 ", pcr amplification is carried out in 72 ℃ of 10 ' 1 circulation, and acquisition contains the amplified production of each protein gene of complete open reading frame sequence.Amplified production is through sequence verification, and the sequence that records with embodiment 1 conforms to, and changes amplified production over to host cell with routine techniques subsequently, to obtain recombinant protein.
Gene specific primer
Clone's title Special primer 1 (5 ' → 3 ') Special primer 2 (5 ' → 3 ')
PP610 GCTTCCTGCTCTCGCCGGAGTTT GCAGGTGTGGACACCAGAGGGCT
PP784 CTTCCCGCAGAGTTGTGAAGCGA ACGCCTAACAAAACCAGCACGCA
PP827 CCTCTGCACGCTAGGGTGGTCCT CCATAAGGGAGACTGTTACATGGGGG
PP1057 GAGAGAAACGGGCTGGGGAGTGA ACTCTGGCAAAGGGAGCCGTGAG
PP1551 GGAAGTGGCTGTTTGGCTGCTGA TGCAGTGTGGATGCTCCTGGATG
PP1553 CAGGTGTGTGTGGGTCCTCCACC TTGAAGGGCCCCAAAACAACAGC
PP1665 ACGCGTCCTGGAACCTGAGTGTG ACACACCCACCCTGGCCATATCC
PP2135 CCTTCTGAGTCTGGGCGGCATCT ATCTCCAGTGGGTGGGGCAACAT
PP2281 TCACTCTGGCTGCTGGGACATCA CCACTTGTCCAAGAGCAGCTGGG
PP2447 CCTCTGCACGCTAGGGTGGTCCT CAAGGGGTCAGTACCTGCTCGGG
PP3111 TGCGAGAGTAGCTGGCCAAGGTG GGACATGGGCGTGTCTTTCCACA
Embodiment 3:cDNA cloned sequence is analyzed
1.PP610
A: nucleotide sequence (SEQ ID NO:1) length: 1853bp
1 CTGAGCCTCT CCGTGCAATG ATTAACCCGG CGGGGCGGCC GGCGCGGGAC
51 CCGAGACGGA GGCGCGGGGC CGGGGCGGGA CCCCGCAGGA CCGCTCGGCT
101 TCCTGCTCTC GCCGGAGTTT CCGCGTAGAG GGCGCATCGC CGGCCCGGGG
151 CCCTTGGTGC GGCGTGGCGC AGGGCGCGGC GTGGGGCGCG CGTGGGCGCG
201 GCGCAGGCGG CCCGGGTCAC CATGAGGACT CTCCGCAGGT TGAAGTTCAT
251 GAGTTCGCCC AGCCTCAGTG ACCTGGGCAA GAGAGAGCCG GCCGCCGCCG
301 CGGACGAGCG GGGCACGCAG CAGCGCCGGG CCTGCGCCAA CGCCACCTGG
351 AACAGCATCC ACAACGGGGT GATCGCCGTC TTCCAGCGCA AGGGGCTGCC
401 CGACCAGGAG CTCTTCAGCC TCAACGAGGG CGTCCGGCAG CTGTTGAAGA
451 CAGAGCTGGG GTCCTTCTTC ACGGAGTACC TGCAGAACCA GCTGCTGACA
501 AAAGGCATGG TGATCCTrCG GGACAAGATT CGCTTCTATG AGGGACAGAA
551 GCTGCTGGAC TCACTGGCAG AGACCTGGGA CTTCTTCTTC AGTGACGTGC
601 TGCCCATGCT GCAGGCCATC TTCTACCCGG TGCAGGGCAA GGAGCCATCG
651 GTGCGCCAGC TGGCCCTGCT GCACTTCCGG AATGCCATCA CCCTCAGTGT
701 GAAGCTAGAG GATGCGCTGG CCCGGGCCCA TGCCCGTGTG CCCCCTGCCA
751 TCGTGCAGAT GCTGCTGGTG CTGCAGGGGG TACATGAGTC CAGGGGCGTG
801 ACTGAGGACT ACCTGCGCCT GGAGACGCTG GTCCAGAAGG TGGTGTCGCC
851 ATACCTGGGC ACCTACGGCC TCCACTCCAG CGAGGGGCCC TTCACCCATT
901 CCTGCATCCT GGAAAAGCGC CTCCTCCGCC GCTCCCGCTC GGGGGACGTG
951 CTGGCCAAGA ACCCTGTGGT GCGCTCCAAG AGCTACAACA CGCCTCTGCT
1001 GAACCCCGTG CAGGAGCACG AGGCGGAGGG CGCGGCGGCC GGCGGTACCA
1051 GCATCCGCAG GCACTCTGTG TCGGAGATGA CGTCCTGCCC CGAGCCTCAG
1101 GGCTTCTCCG ACCCGCCCGG CCAGGGCCCC ACCGGGACCT TCAGGTCCTC
1151 CCCGGCGCCC CACTCAGGGC CCTGCCCCAG CAGACTGTAC CCCACGACCC
1201 AGCCCCCTGA GCAGGGCTTG GATCCCACCC GCAGCTCCCT GCCCCGCTCC
1251 AGCCCGGAGA ACCTGGTGGA CCAGATCCTG GAGTCCGTGG ACTCGGATTC
1301 TGAAGGGATT TTCATTGACT TTGGCCGGGG CCGGGGCTCT GGCATGTCCG
1351 ACTTGGAGGG CTCTGGGGGC CGGCAGAGTG TCGTGTGAGG CCTCACAGCT
1401 GGCCTTGAGT TTTTACTGAC ACGTCCCTGT GTGCGGGGGT GTCCATGTGG
1451 CGTGTGTGTG AGTGAGACTT TTTTACTGCG TCCCGTCCCG CCAGCCCTAT
1501 CGGCCTCGTC ACTGGCCTTG GTCACTTTGT ATTTCTGTCT TGGTTGGAAA
1551 TACCATCAGC CTTCCTTGCT CGGCCCAGGT CTGTTTCAGG CATCTGAGTC
1601 GGCGTTTACC CAGGGGCCGG GCCAGAGACG GGGGTCGGCC GCTCGCTCCC
1651 ACGCTCCTCC TGCCCCAGCC CTCTGGTGTC CACACCTGCC CACAGAGAAT
1701 GTAAACCCAG TGGGCTCTGC CCACGCCGGG CCCCAAAGTG ACCAGACTCC
1751 AGCACACCTG TCTCCTCCTG CCTGGGGTGG CCATGGGGAT GGAAGGGGGT
1801 GGAATAAAAC CTGTCACCCT GGGAAAAAAA AAAAAAAAAA AAAAAAAAAA
1851 AAA
B: aminoacid sequence (SEQ ID NO:2) length: 388 amino acid
1 MRTLRRLKFM SSPSLSDLGK REPAAAADER GTQQRRACAN ATWNSIHNGV
51 IAVFQRKGLP DQELFSLNEG VRQLLKTELG SFFTEYLQNQ LLTKGMVILR
101 DKIRFYEGQK LLDSLAETWD FFFSDVLPML QAIFYPVQGK EPSVRQLALL
151 HFRNAITLSV KLEDALARAH ARVPPAIVQM LLVLQGVHES RGVTEDYLRL
201 ETLVQKVVSP YLGTYGLHSS EGPFTHSCIL EKRLLRRSRS GDVLAKNPVV
251 RSKSYNTPLL NPVQEHEAEG AAAGGTSIRR HSVSEMTSCP EPQGFSDPPG
301 QGPTGTFRSS PAPHSGPCPS RLYPTTQPPE QGLDPTRSSL PRSSPENLVD
351 QILESVDSDS EGIFIDFGRG RGSGMSDLEG SGGRQSVV
C. Nucleotide and amino acid composite sequence (SEQ ID NO:3)
Clone number and protein name: PP610
Start code: 222 ATG stop coding: 1388 TGA
Protein molecular weight: 42751.17
1 CT GAG CCT CTC CGT GCA ATG ATT AAC CCG GCG GGG CGG CCG GCG CGG 47
48 GAC CCG AGA CGG AGG CGC GGG GCC GGG GCG GGA CCC CGC AGG ACC GCT 95
96 CGG CTT CCT GCT CTC GCC GGA GTT TCC GCG TAG AGG GCG CAT CGC CGG 143
144 CCC GGG GCC CTT GGT GCG GCG TGG CGC AGG GCG CGG CGT GGG GCG CGC 191
192 GTG GGC GCG GCG CAG GCG GCC CGG GTC ACC ATG AGG ACT CTC CGC AGG 239
1 Met Arg Thr Leu Arg Arg 6
240 TTG AAG TTC ATG AGT TCG CCC AGC CTC AGT GAC CTG GGC AAG AGA GAG 287
7 Leu Lys Phe Met Ser Ser Pro Ser Leu Ser Asp Leu Gly Lys Arg Glu 22
288 CCG GCC GCC GCC GCG GAC GAG CGG GGC ACG CAG CAG CGC CGG GCC TGC 335
23 Pro Ala Ala Ala Ala Asp Glu Arg Gly Thr Gln Gln Arg Arg Ala Cys 38
336 GCC AAC GCC ACC TGG AAC AGC ATC CAC AAC GGG GTG ATC GCC GTC TTC 383
39 Ala Asn Ala Thr Trp Asn Ser Ile His Asn Gly Val Ile Ala Val Phe 54
384 CAG CGC AAG GGG CTG CCC GAC CAG GAG CTC TTC AGC CTC AAC GAG GGC 431
55 Gln Arg Lys Gly Leu Pro Asp Gln Glu Leu Phe Ser Leu Asn Glu Gly 70
432 GTC CGG CAG CTG TTG AAG ACA GAG CTG GGG TCC TTC TTC ACG GAG TAC 479
71 Val Arg Gln Leu Leu Lys Thr Glu Leu Gly Ser Phe Phe Thr Glu Tyr 86
480 CTG CAG AAC CAG CTG CTG ACA AAA GGC ATG GTG ATC CTT CGG GAC AAG 527
87 Leu Gln Asn Gln Leu Leu Thr Lys Gly Met Val Ile Leu Arg Asp Lys 102
528 ATT CGC TTC TAT GAG GGA CAG AAG CTG CTG GAC TCA CTG GCA GAG ACC 575
103 Ile Arg Phe Tyr Glu Gly Gln Lys Leu Leu Asp Ser Leu Ala Glu Thr 118
576 TGG GAC TTC TTC TTC AGT GAC GTG CTG CCC ATG CTG CAG GCC ATC TTC 623
119 Trp Asp Phe Phe Phe Ser Asp Val Leu Pro Met Leu Gln Ala Ile Phe 134
624 TAC CCG GTG CAG GGC AAG GAG CCA TCG GTG CGC CAG CTG GCC CTG CTG 671
135 Tyr Pro Val Gln Gly Lys Glu Pro Ser Val Arg Gln Leu Ala Leu Leu 150
672 CAC TTC CGG AAT GCC ATC ACC CTC AGT GTG AAG CTA GAG GAT GCG CTG 719
151 His Phe Arg Asn Ala Ile Thr Leu Ser Val Lys Leu Glu Asp Ala Leu 166
720 GCC CGG GCC CAT GCC CGT GTG CCC CCT GCC ATC GTG CAG ATG CTG CTG 767
167 Ala Arg Ala His Ala Arg Val Pro Pro Ala Ile Val Gln Met Leu Leu 182
768 GTG CTG CAG GGG GTA CAT GAG TCC AGG GGC GTG ACT GAG GAC TAC CTG 815
183 Val Leu Gln Gly Val His Glu Ser Arg Gly Val Thr Glu Asp Tyr Leu 198
816 CGC CTG GAG ACG CTG GTC CAG AAG GTG GTG TCG CCA TAC CTG GGC ACC 863
199 Arg Leu Glu Thr Leu Val Gln Lys Val Val Ser Pro Tyr Leu Gly Thr 214
864 TAC GGC CTC CAC TCC AGC GAG GGG CCC TTC ACC CAT TCC TGC ATC CTG 911
215 Tyr Gly Leu His Ser Ser Glu Gly Pro Phe Thr His Ser Cys Ile Leu 230
912 GAA AAG CGC CTC CTC CGC CGC TCC CGC TCG GGG GAC GTG CTG GCC AAG 959
231 Glu Lys Arg Leu Leu Arg Arg Ser Arg Ser Gly Asp Val Leu Ala Lys 246
960 AAC CCT GTG GTG CGC TCC AAG AGC TAC AAC ACG CCT CTG CTG AAC CCC 1007
247 Asn Pro Val Val Arg Ser Lys Ser Tyr Asn Thr Pro Leu Leu Asn Pro 262
1008 GTG CAG GAG CAC GAG GCG GAG GGC GCG GCG GCC GGC GGT ACC AGC ATC 1055
263 Val Gln Glu His Glu Ala Glu Gly Ala Ala Ala Gly Gly Thr Ser Ile 278
1056 CGC AGG CAC TCT GTG TCG GAG ATG ACG TCC TGC CCC GAG CCT CAG GGC 1103
279 Arg Arg His Ser Val Ser Glu Met Thr Ser Cys Pro Glu Pro Gln Gly 294
1104 TTC TCC GAC CCG CCC GGC CAG GGC CCC ACC GGG ACC TTC AGG TCC TCC 1151
295 Phe Ser Asp Pro Pro Gly Gln Gly Pro Thr Gly Thr Phe Arg Ser Ser 310
1152 CCG GCG CCC CAC TCA GGG CCC TGC CCC AGC AGA CTG TAC CCC ACG ACC 1199
311 Pro Ala Pro His Ser Gly Pro Cys Pro Ser Arg Leu Tyr Pro Thr Thr 326
1200 CAG CCC CCT GAG CAG GGC TTG GAT CCC ACC CGC AGC TCC CTG CCC CGC 1247
327 Gln Pro Pro Glu Gln Gly Leu Asp Pro Thr Arg Ser Ser Leu Pro Arg 342
1248 TCC AGC CCG GAG AAC CTG GTG GAC CAG ATC CTG GAG TCC GTG GAC TCG 1295
343 Ser Ser Pro Glu Asn Leu Val Asp Gln Ile Leu Glu Ser Val Asp Ser 358
1296 GAT TCT GAA GGG ATT TTC ATT GAC TTT GGC CGG GGC CGG GGC TCT GGC 1343
359 Asp Ser Glu Gly Ile Phe Ile Asp Phe Gly Arg Gly Arg Gly Ser Gly 374
1344 ATG TCC GAC TTG GAG GGC TCT GGG GGC CGG CAG AGT GTC GTG TGA GGC 1391
375 Met Ser Asp Leu Glu Gly Ser Gly Gly Arg Gln Ser Val Val *** 389
1392 CTC ACA GCT GGC CTT GAG TTT TTA CTG ACA CGT CCC TGT GTG CGG GGG 1439
1440 TGT CCA TGT GGC GTG TGT GTG AGT GAG ACT TTT TTA CTG CGT CCC GTC 1487
1488 CCG CCA GCC CTA TCG GCC TCG TCA CTG GCC TTG GTC ACT TTG TAT TTC 1535
1536 TGT CTT GGT TGG AAA TAC CAT CAG CCT TCC TTG CTC GGC CCA GGT CTG 1583
1584 TTT CAG GCA TCT GAG TCG GCG TTT ACC CAG GGG CCG GGC CAG AGA CGG 1631
1632 GGG TCG GCC GCT CGC TCC CAC GCT CCT CCT GCC CCA GCC CTC TGG TGT 1679
1680 CCA CAC CTG CCC ACA GAG AAT GTA AAC CCA GTG GGC TCT GCC CAC GCC 1727
1728 GGG CCC CAA AGT GAC CAG ACT CCA GCA CAC CTG TCT CCT CCT GCC TGG 1775
1776 GGT GGC CAT GGG GAT GGA AGG GGG TGG AAT AAA ACC TGT CAC CCT GGG 1823
1824 AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA 1853
D:Blas+p homology comparative result
Query=pp610[gene=pp610] (388 character)
>SP_HUM：O75982 O75982 homo sapiens(human).hypothetical 18.0kd
Protein (fragment) .11/1998 length=158
Score value=316bits (802), desired value=2e-85
Homogeny=158/158 (100%), similarity=158/158 (100%)
Query：73 QLLKTELGSFFTEYLQNQLLTKGMVILRDKIRFYEGQKLLDSLAETWDFFFSDVLPMLQA 132
QLLKTELGSFFTEYLQNQLLTKGMVILRDKIRFYEGQKLLDSLAETWDFFFSDVLPMLQA
Sbjct：1 QLLKTELGSFFTEYLQNQLLTKGMVILRDKIRFYEGQKLLDSLAETWDFFFSDVLPMLQA 60
Query：133 IFYPVQGKEPSVRQLALLHFRNAITLSVKLEDALARAHARVPPAIVQMLLVLQGVHESRG 192
IFYPVQGKEPSVRQLALLHFRNAITLSVKLEDALARAHARVPPAIVQMLLVLQGVHESRG
Sbjct：61 IFYPVQGKEPSVRQLALLHFRNAITLSVKLEDALARAHARVPPAIVQMLLVLQGVHESRG 120
Query：193 VTEDYLRLETLVQKVVSPYLGTYGLHSSEGPFTHSCIL 230
VTEDYLRLETLVQKVVSPYLGTYGLHSSEGPFTHSCIL
Sbjct：121 VTEDYLRLETLVQKVVSPYLGTYGLHSSEGPFTHSCIL 158
2.PP784
A: nucleotide sequence (SEQ ID NO:4) length: 2145bp
1 GGCGATTCTT CCCGCAGAGT TGTGAAGCGA AAGGCTTACA ATTAAAAGGA
51 AGAAAAAAAA ATAAAGATAA TTCGGGAGTA CAATTGACAA AGCGTGTGGG
101 TCGCTCAGCC TCCAGCAGTA ACTGCTGATC TCCAGTTCTT GGAGGGTTCC
151 GGTGAGAAGA ACGCCCCTAC TGCGGTACTG AGGAAGCGGC AGGAGGAGAT
201 GCGGCCCCTG GACATAGACG AGGTGGAAGC GCCTGAGGAA GTGGAGGTGC
251 TGGAGCCCGA GGAGGATTTC GAGCAGTTCC TGCTCCCGGT CATCAACGAG
301 ATGCGCGAGG ACATCGCGTC TCTTATACGC GAGCACGGGC GGGCGTACCT
351 GCGGACCAGG AGCAAGCTGT GGGAGATGGA CAATATGCTT ATCCAGATCA
401 AAACGCAGGT GGAGGCCTCG GAGGAGAGCG CCCTCAATCA CGTGCAGCAC
451 CCGAGTGGCG AAGCCGACGA GAGAGTGTCG GAGTTGTGCG AGAAGGCTGA
501 GGAGAAAGCC AAGGAGATTG CGAAGATGGC AGAGATGCTG GTCGAGCTCG
551 TCTGGCGAAT AGAGAGAAGC GAGTCTTCTT GAAGGAGGAG ATCGGTGGTA
601 AGGTCGGAAA CTAGCTGGAG CTTGGATGAA ACTCAGTAGT CGCCCTAAGC
651 ATGGTTAAAC CGCGCCCTGT AAAAAGTTAT GTTCTAGGCC TGCATCCCCC
701 ATTTTATTGC ATCAAAAATT GAGCATTGGG AACAAAGTTG GGGTCAAGAG
751 GAAAGAATGC GTGCTGGTTT TGTTAGGCGT TAGTATACCG TTTTTTTTGT
801 GGCCTCTCCC TCCCACACTG GTAATTAGAG AAAGATAACA GTAACTTCGG
851 TTTAGTTTTT GTGAAACATA AAAGTCAATT CTAATAGGGC AGTCGCCAGA
901 AGTAGACCTG TCTAGGCACT AAGGGAGTTT GGGGAAAGCC AAAGAAGACC
951 TAGGCCATAG AGCACAGTGG AACGCAGGTG AGAACGCAGG GAAAGAGAAG
1001 TAAAGAGTAA AGCCAGAGGC CATTACCTGA AATTTCCAGA TTGTTCTATG
1051 AGACAGGTAT GTCAGAGGAC CGTGTCTCAA AGAAGTGGCA TTCTTCTGGG
1101 TGGTTCACAT TTATTTTTAA CATCAGGATA TGGAACTTTG AGAGTCAATG
1151 TCTTCATGGT AAATTTCTCC CAATTGTGCT TTAGGTTCAC AGCTGAGGAC
1201 TGTAGTCAAC TGGTAAGGAT GAACTGACAC CTTGAGGAAA CTGAAACAGC
1251 TTGTCATTTT CTCTGTTTTG TTTTTGTATG TTTTTTACCC CCATGTAACA
1301 GTCTCCCTTA TGGTCAGTGA TTGATGACCT CGATATGGAT TTAGATTATC
1351 AAATGTGTTT GGTTTTGGAA ATATAACTTT TGTCAAAGAA ATACTCTCAG
1401 AAGAGAAATG GGGCTTAATT AAGTTGTTTT TGTGGTCACG TTTATTCTTG
1451 TTACTTCGCT GTGTTTTTGA AATGTTGGGC ATGGCCTCGT ATTTTGCTGT
1501 TACCTTTGTG ACCTGATTGT TTTTTGGAAC ACGTCAAGAC GTGGGATCAG
1551 AATCTTCCAA CTTTAGAGGT GCAATGGAAG ACACTACGCT ACTTGGTTGA
1601 GCCTGGTGAA GAATGTATTA ATGAGACTGC TTTGCATAAA ACTGGGAAGA
1651 AAGAGAAGAC AGTTGGAGAT GGAAGATGGT TTTGTATATA TTTTGGAACT
1701 TTAGTTCCTC TGTGAGACGA AAGAGGAGAG CTATGTTTTG TGTCACATTG
1751 TCTGATATAT ATTGTGTAAC CTGTCAGGTG AGTTGATTTA GACAACATAG
1801 CTGACCTTTT ATGACAAGGC AGTTTGAATA GGGACTATTG TAATACCCTC
1851 ACACATTATA GGGGCATCAG AGAATGGCAT GGAAGAGACA GTCTACAGAG
1901 AGCTTTAAGA GGCCGGAGAA AGGAAAAGAC ATTATCAGGG CCTGGAAAGT
1951 CTCTTCCAGT TCATCAGGGT AGTAGACCTG TCAGATTGGA GGTCAAAGTC
2001 AAAAGTATAA TAAGTGTTAA TTTGCCTTGA AAAACAAAGA CCTAAAAAGC
2051 CTTTTTTAAG AAATAAATTT TTTGTTCACA TGACCAAAGC CCCTCCTGTG
2101 TGTGTTAATA AAAGAATCAT AAACAAAAAA AAAAAAAAAA AAAAA
B: aminoacid sequence (SEQ ID NO:5) length: 127 amino acid
1 MRPLDIDEVE APEEVEVLEP EEDFEQFLLP VINEMREDIA SLIREHGRAY
51 LRTRSKLWEM DNMLIQIKTQ VEASEESALN HVQHPSGEAD ERVSELCEKA
101 EEKAKEIAKM AEMLVELVWR IERSESS
C. Nucleotide and amino acid composite sequence (SEQ ID NO:6)
Clone number and protein name: PP784
Start code: 199 ATG stop coding: 582 TGA
Protein molecular weight: 14806.89
1 GGC GAT TCT TCC CGC AGA GTT GTG AAG CGA AAG GCT TAC AAT TAA AAG 48
49 GAA GAA AAA AAA ATA AAG ATA ATT CGG GAG TAC AAT TGA CAA AGC GTG 96
97 TGG GTC GCT CAG CCT CCA GCA GTA ACT GCT GAT CTC CAG TTC TTG GAG 144
145 GGT TCC GGT GAG AAG AAC GCC CCT ACT GCG GTA CTG AGG AAG CGG CAG 192
193 GAG GAG ATG CGG CCC CTG GAC ATA GAC GAG GTG GAA GCG CCT GAG GAA 240
1 Met Arg Pro Leu Asp Ile Asp Glu Val Glu Ala Pro Glu Glu 14
241 GTG GAG GTG CTG GAG CCC GAG GAG GAT TTC GAG CAG TTC CTG CTC CCG 288
15 Val Glu Val Leu Glu Pro Glu Glu Asp Phe Glu Gln Phe Leu Leu Pro 30
289 GTC ATC AAC GAG ATG CGC GAG GAC ATC GCG TCT CTT ATA CGC GAG CAC 336
31 Val Ile Asn Glu Met Arg Glu Asp Ile Ala Ser Leu Ile Arg Glu His 46
337 GGG CGG GCG TAC CTG CGG ACC AGG AGC AAG CTG TGG GAG ATG GAC AAT 384
47 Gly Arg Ala Tyr Leu Arg Thr Arg Ser Lys Leu Trp Glu Met Asp Asn 62
385 ATG CTT ATC CAG ATC AAA ACG CAG GTG GAG GCC TCG GAG GAG AGC GCC 432
63 Met Leu Ile Gln Ile Lys Thr Gln Val Glu Ala Ser Glu Glu Ser Ala 78
433 CTC AAT CAC GTG CAG CAC CCG AGT GGC GAA GCC GAC GAG AGA GTG TCG 480
79 Leu Asn His Val Gln His Pro Ser Gly Glu Ala Asp Glu Arg Val Ser 94
481 GAG TTG TGC GAG AAG GCT GAG GAG AAA GCC AAG GAG ATT GCG AAG ATG 528
95 Glu Leu Cys Glu Lys Ala Glu Glu Lys Ala Lys Glu Ile Ala Lys Met 110
529 GCA GAG ATG CTG GTC GAG CTC GTC TGG CGA ATA GAG AGA AGC GAG TCT 576
111 Ala Glu Met Leu Val Glu Leu Val Trp Arg Ile Glu Arg Ser Glu Ser 126
577 TCT TGA AGG AGG AGA TCG GTG GTA AGG TCG GAA ACT AGC TGG AGC TTG 624
127 Ser *** 128
625 GAT GAA ACT CAG TAG TCG CCC TAA GCA TGG TTA AAC CGC GCC CTG TAA 672
673 AAA GTT ATG TTC TAG GCC TGC ATC CCC CAT TTT ATT GCA TCA AAA ATT 720
721 GAG CAT TGG GAA CAA AGT TGG GGT CAA GAG GAA AGA ATG CGT GCT GGT 768
769 TTT GTT AGG CGT TAG TAT ACC GTT TTT TTT GTG GCC TCT CCC TCC CAC 816
817 ACT GGT AAT TAG AGA AAG ATA ACA GTA ACT TCG GTT TAG TTT TTG TGA 864
865 AAC ATA AAA GTC AAT TCT AAT AGG GCA GTC GCC AGA AGT AGA CCT GTC 912
913 TAG GCA CTA AGG GAG TTT GGG GAA AGC CAA AGA AGA CCT AGG CCA TAG 960
961 AGC ACA GTG GAA CGC AGG TGA GAA CGC AGG GAA AGA GAA GTA AAG AGT 1008
1009 AAA GCC AGA GGC CAT TAC CTG AAA TTT CCA GAT TGT TCT ATG AGA CAG 1056
1057 GTA TGT CAG AGG ACC GTG TCT CAA AGA AGT GGC ATT CTT CTG GGT GGT 1104
1105 TCA CAT TTA TTT TTA ACA TCA GGA TAT GGA ACT TTG AGA GTC AAT GTC 1152
1153 TTC ATG GTA AAT TTC TCC CAA TTG TGC TTT AGG TTC ACA GCT GAG GAC 1200
1201 TGT AGT CAA CTG GTA AGG ATG AAC TGA CAC CTT GAG GAA ACT GAA ACA 1248
1249 GCT TGT CAT TTT CTC TGT TTT GTT TTT GTA TGT TTT TTA CCC CCA TGT 1296
1297 AAC AGT CTC CCT TAT GGT CAG TGA TTG ATG ACC TCG ATA TGG ATT TAG 1344
1345 ATT ATC AAA TGT GTT TGG TTT TGG AAA TAT AAC TTT TGT CAA AGA AAT 1392
1393 ACT CTC AGA AGA GAA ATG GGG CTT AAT TAA GTT GTT TTT GTG GTC ACG 1440
1441 TTT ATT CTT GTT ACT TCG CTG TGT TTT TGA AAT GTT GGG CAT GGC CTC 1488
1489 GTA TTT TGC TGT TAC CTT TGT GAC CTG ATT GTT TTT TGG AAC ACG TCA 1536
1537 AGA CGT GGG ATC AGA ATC TTC CAA CTT TAG AGG TGC AAT GGA AGA CAC 1584
1585 TAC GCT ACT TGG TTG AGC CTG GTG AAG AAT GTA TTA ATG AGA CTG CTT 1632
1633 TGC ATA AAA CTG GGA AGA AAG AGA AGA CAG TTG GAG ATG GAA GAT GGT 1680
1681 TTT GTA TAT ATT TTG GAA CTT TAG TTC CTC TGT GAG ACG AAA GAG GAG 1728
1729 AGC TAT GTT TTG TGT CAC ATT GTC TGA TAT ATA TTG TGT AAC CTG TCA 1776
1777 GGT GAG TTG ATT TAG ACA ACA TAG CTG ACC TTT TAT GAC AAG GCA GTT 1824
1825 TGA ATA GGG ACT ATT GTA ATA CCC TCA CAC ATT ATA GGG GCA TCA GAG 1872
1873 AAT GGC ATG GAA GAG ACA GTC TAC AGA GAG CTT TAA GAG GCC GGA GAA 1920
1921 AGG AAA AGA CAT TAT CAG GGC CTG GAA AGT CTC TTC CAG TTC ATC AGG 1968
1969 GTA GTA GAC CTG TCA GAT TGG AGG TCA AAG TCA AAA GTA TAA TAA GTG 2016
2017 TTA ATT TGC CTT GAA AAA CAA AGA CCT AAA AAG CCT TTT TTA AGA AAT 2064
2065 AAA TTT TTT GTT CAC ATG ACC AAA GCC CCT CCT GTG TGT GTT AAT AAA 2112
2113 AGA ATC ATA AAC AAA AAA AAA AAA AAA AAA AAA 2145
3.PP827
A: nucleotide sequence (SEQ ID NO:7) length: 1678bp
1 CCATGTTGGC CAGGCTGGTC TGAGAGAGAA CTCCTGACCT CGTGATCCGC
51 CTGCCTCAGC CTCCCAAAGT GCTGGGATGA CAGGCGTGCG CCACTGCACC
101 CGGCCCGTGA AGGTCTTTTC TGAAGGGAGC TACCTTTGAG AAGTGCCTGT
151 TGGGACAGAA CTCCCTGCTC TGGTCTGACC CTCTGCACGC TAGGGTGGTC
201 CTGCACCTGT CGGGGAGGGG GCCAAGGACA CAGCACTGGG CTGGGGCAGG
251 GCTCTGCGCT GGTGCTGCCT GCCATCCCAG TTCTGCCATT CCCCCAGCGC
301 ACCCCCGCAC CCGCCCCTCC AGCTGATGCC TGCTCCCTCT CTCTGCAGAA
351 CGGGCTCATG TCGGGGCTGA TGCAGATGCT GCTGCTGAAG GTGTCTGCAC
401 ACATCACCGA GCAGCTGGGC ATGGCCCCAG GTGGCGAGTT CAGGGAGGCC
451 TTCAAGGAGG TGGGCACAGG GTGAGGTAGG GGGTGGCCAC AGGGATGTGG
501 CTCACAGGGG TGGTCCGGGG TTCCCAGTGC TCCCCAACAC CCAGCCTCTG
551 CTCCAGGCCA GCAAGGTGCC TTTCTGCAAG TTCCACCTGG GTGACCGACC
601 CATCCCCGTC ACCTTCAAGA GGGCCATCGC AGCGCTCTCC TTCTGGCAGA
651 AGGTCAGGCT GGCTTGGGGC CTGTGCTTCC TGTCAGACCC CATCAGGTAG
701 GGCTGCCCCC GGGACCCTGG CCGGCCTGCA GGGTGGTCTG TGGGAGGCTC
751 CAGGCCCTCC TGTGCAGGTC CAAGCGCAGC CAATCCTCAC TCAAGGCCTT
801 CCCTGCCCTT TCCTTCCGCC ACAAATCCCA AACAAACGTG CTGTGGTCCC
851 TGCCCGGTGT CCACAGTGCC AGCCCCACCC CCCCAGCCCG TTGCCCATCC
901 CTGCGGGGCT GCAGCCATCC CTCTCCACAG CAAGGATGAC GTGGAACGCT
951 GCAAGCAGAA GGACCTACTG GAGCAGATGA TGGCCGAGAT GATTGGCGAG
1001 TTCCCAGACC TGCACCGCAC CATCGTCTCG GAGCGCGACG TCTACCTAAC
1051 CTACATGCTG CGCCAGGCCG CGCGGCGCCT CGAGCTGCCT CGGGCCTCTG
1101 ACGGTGACGG CCGCCCGCAG GCGTGGGACC CCCTGTGAGG CTGAGGCCCG
1151 AGCAGGTACT GACCCCTTGT CCTTCCCCAC AGCCGAGCCC AGGAAGTGCG
1201 TCCCCTCCGT GGTCGTGGGC GTCGTGGGCA TGGGCCACGT GCCTGGCATC
1251 GAGAAGAACT GGAGCACCGA CCTCAACATC CAGGAGATCA TGACCGTGCC
1301 CCCGCCGTCC GTCTCCGGCA GAGTGTCTCG GTTGGCCGTG AAGGCCGCCT
1351 TCTTCGGCCT GCTGGGCTAC AGCCTGTACT GGATGGGCCG CCGCACCGCG
1401 AGCCTGGTCC TGTCGCTGCC CGCCGCGCAG TACTGCCTGC AGAGGGTGAC
1451 CGAGGCCCGG CACAAGTAGG AGACTGCTCC CCGCCCGCTC GGGCCCCTGA
1501 GGAGCCAGTG CCCCCGCGGC ACTTCTGGGT GCCAGGTGCA TCCTAGCCCG
1551 CCCGAGGCCC CTGCCACCCC CCATGGGGGT CTGGGCCCGG CCTCGCCTGC
1601 CCTCCTGGGC CAGTCACCCC TCCCCCAGCC CACCCAAATA AAGGATTATT
1651 TAACTGTCTG AAAAAAAAAA AAAAAAAA
B: aminoacid sequence (SEQ ID NO:8) length: 161 amino acid
1 MWLTGVVRGS QCSPTPSLCS RPARCLSASS TWVTDPSPSP SRGPSQRSPS
51 GRRSGWLGAC ASCQTPSGRA APGTLAGLQG GLWEAPGPPV QVQAQPILTQ
101 GLPCPFLPPQ IPNKRAVVPA RCPQCQPHPP SPLPIPAGLQ PSLSTARMTW
151 NAASRRTYWS R
C. Nucleotide and amino acid composite sequence (SEQ ID NO:9)
Clone number and protein name: PP827
Start code: 495 ATG stop coding: 980 TGA
Protein molecular weight: 16922.45
1 CC ATG TTG GCC AGG CTG GTC TGA GAG AGA ACT CCT GAC CTC GTG ATC 47
48 CGC CTG CCT CAG CCT CCC AAA GTG CTG GGA TGA CAG GCG TGC GCC ACT 95
96 GCA CCC GGC CCG TGA AGG TCT TTT CTG AAG GGA GCT ACC TTT GAG AAG 143
144 TGC CTG TTG GGA CAG AAC TCC CTG CTC TGG TCT GAC CCT CTG CAC GCT 191
192 AGG GTG GTC CTG CAC CTG TCG GGG AGG GGG CCA AGG ACA CAG CAC TGG 239
240 GCT GGG GCA GGG CTC TGC GCT GGT GCT GCC TGC CAT CCC AGT TCT GCC 287
288 ATT CCC CCA GCG CAC CCC CGC ACC CGC CCC TCC AGC TGA TGC CTG CTC 335
336 CCT CTC TCT GCA GAA CGG GCT CAT GTC GGG GCT GAT GCA GAT GCT GCT 383
384 GCT GAA GGT GTC TGC ACA CAT CAC CGA GCA GCT GGG CAT GGC CCC AGG 431
432 TGG CGA GTT CAG GGA GGC CTT CAA GGA GGT GGG CAC AGG GTG AGG TAG 479
480 GGG GTG GCC ACA GGG ATG TGG CTC ACA GGG GTG GTC CGG GGT TCC CAG 527
1 Met Trp Leu Thr Gly Val Val Arg Gly Ser Gln 11
528 TGC TCC CCA ACA CCC AGC CTC TGC TCC AGG CCA GCA AGG TGC CTT TCT 575
12 Cys Ser Pro Thr Pro Ser Leu Cys Ser Arg Pro Ala Arg Cys Leu Ser 27
576 GCA AGT TCC ACC TGG GTG ACC GAC CCA TCC CCG TCA CCT TCA AGA GGG 623
28 Ala Ser Ser Thr Trp Val Thr Asp Pro Ser Pro Ser Pro Ser Arg Gly 43
624 CCA TCG CAG CGC TCT CCT TCT GGC AGA AGG TCA GGC TGG CTT GGG GCC 671
44 Pro Ser Gln Arg Ser Pro Ser Gly Arg Arg Ser Gly Trp Leu Gly Ala 59
672 TGT GCT TCC TGT CAG ACC CCA TCA GGT AGG GCT GCC CCC GGG ACC CTG 719
60 Cys Ala Ser Cys Gln Thr Pro Ser Gly Arg Ala Ala Pro Gly Thr Leu 75
720 GCC GGC CTG CAG GGT GGT CTG TGG GAG GCT CCA GGC CCT CCT GTG CAG 767
76 Ala Gly Leu Gln Gly Gly Leu Trp Glu Ala Pro Gly Pro Pro Val Gln 91
768 GTC CAA GCG CAG CCA ATC CTC ACT CAA GGC CTT CCC TGC CCT TTC CTT 815
92 Val Gln Ala Gln Pro Ile Leu Thr Gln Gly Leu Pro Cys Pro Phe Leu 107
816 CCG CCA CAA ATC CCA AAC AAA CGT GCT GTG GTC CCT GCC CGG TGT CCA 863
108 Pro Pro Gln Ile Pro Asn Lys Arg Ala Val Val Pro Ala Arg Cys Pro 123
864 CAG TGC CAG CCC CAC CCC CCC AGC CCG TTG CCC ATC CCT GCG GGG CTG 911
124 Gln Cys Gln Pro His Pro Pro Ser Pro Leu Pro Ile Pro Ala Gly Leu 139
912 CAG CCA TCC CTC TCC ACA GCA AGG ATG ACG TGG AAC GCT GCA AGC AGA 959
140 Gln Pro Ser Leu Ser Thr Ala Arg Met Thr Trp Asn Ala Ala Ser Arg 155
960 AGG ACC TAC TGG AGC AGA TGA TGG CCG AGA TGA TTG GCG AGT TCC CAG 1007
156 Arg Thr Tyr Trp Ser Arg *** 162
1008 ACC TGC ACC GCA CCA TCG TCT CGG AGC GCG ACG TCT ACC TAA CCT ACA 1055
1056 TGC TGC GCC AGG CCG CGC GGC GCC TCG AGC TGC CTC GGG CCT CTG ACG 1103
1104 GTG ACG GCC GCC CGC AGG CGT GGG ACC CCC TGT GAG GCT GAG GCC CGA 1151
1152 GCA GGT ACT GAC CCC TTG TCC TTC CCC ACA GCC GAG CCC AGG AAG TGC 1199
1200 GTC CCC TCC GTG GTC GTG GGC GTC GTG GGC ATG GGC CAC GTG CCT GGC 1247
1248 ATC GAG AAG AAC TGG AGC ACC GAC CTC AAC ATC CAG GAG ATC ATG ACC 1295
1296 GTG CCC CCG CCG TCC GTC TCC GGC AGA GTG TCT CGG TTG GCC GTG AAG 1343
1344 GCC GCC TTC TTC GGC CTG CTG GGC TAC AGC CTG TAC TGG ATG GGC CGC 1391
1392 CGC ACC GCG AGC CTG GTC CTG TCG CTG CCC GCC GCG CAG TAC TGC CTG 1439
1440 CAG AGG GTG ACC GAG GCC CGG CAC AAG TAG GAG ACT GCT CCC CGC CCG 1487
1488 CTC GGG CCC CTG AGG AGC CAG TGC CCC CGC GGC ACT TCT GGG TGC CAG 1535
1536 GTG CAT CCT AGC CCG CCC GAG GCC CCT GCC ACC CCC CAT GGG GGT CTG 1583
1584 GGC CCG GCC TCG CCT GCC CTC CTG GGC CAG TCA CCC CTC CCC CAG CCC 1631
1632 ACC CAA ATA AAG GAT TAT TTA ACT GTC TGA AAA AAA AAA AAA AAA AA 1678
4.PP1057
A: nucleotide sequence (SEQ ID NO:10) length: 2030bp
1 GGCACAGAGC TGGGCATGGG AGAGAAACGG GCTGGGGAGT GAGGCCAGGA
51 GTGGGATTCA GCTGCAGCAG GGCGCCCCCT CCAAACTGCA GCTGGTCTGG
101 CTTACTGTTT TGCCGTTCAA AAAGGTCGCG AATCCGTGGG ACTGAGCACG
151 GGGACCTCAC CCGCTAGCCA GCGTCTGCTG CACTTGATCA GGTGGGGCCT
201 TGGTGGGCGG CTGCCTTTCC TATACAGTTT GTCTTGTCAC CCTGGTTTCC
251 CACTGGGGCC AGGTCTCTTC TCCAGCCTCC ACCTGCCTGT CTGATCCAAG
301 AGCTGAGACA CGGCCACCCA GCACCAGTCA CTCCTCTGTT CACCTTAAGT
351 AACACACAAA TCGGGAACAG GAGGACAGAA CCGTTGGCAT TATCAGGATT
401 CGTGTTTTGT GGGGGTGGGA GTGGAGAGTA GGGTGGTCTT GTGAGTTGTG
451 CAGGGTGAAG ACCGCTTCCC TGAGACAGGG GCAGTGGTGC TGATGGAATG
501 TGGGGGAGGC CCACATTTGA GCAAAGCTGC CCTGCCCTTG TCCCCTGGCC
551 TGGCTTCCTG GTAAGGAGTT TCAGCCGCCT CCGCAGGAAC CCCCAAAGTG
601 CAGATTCCGG AGCAGACACA TCCGGGCGGA GAGACTCAGC AGACAAGTGC
651 TGCAGTTGCA CGGTGGGACC CGGGGCCTCG TGCGTTTTTT GCTGTGGGTG
701 GGGTGGGTGG GTTGGTTTAT GCCTATCAAT GCAATTTTTA ATTTTTGTTA
751 ATATCAACAG CAAAAGCCTA GTGCATTGGG AGATGTGCAA CCTCCCTGAA
801 AATCTTTTCT GTTTCTGGAG TACTTCAGGG GTGGCCTCTG GCCCCAGAGC
851 CTTTGCCACA GTGCTCCCAC CAGCCCCCAC CTCATCCGTC TGTTTGCAGA
901 GCCTCATCTA CAGGTCCCCA CGCTGCCTTC TTTACTCACT CTGCGCTTGG
951 CCGTTTTGTT ATTTGGCTTA GTCTACATTG GGCGGAAGTC TGTGTGCACA
1001 GAGTGGGTGT TCCTTCGAGC CCCTTCCACT CAGAGGGCCA CACCCAGCGA
1051 TGCCAGTGAA GGTGGCACAG CCTCTCTTCA GTTTCTCCTG ACTGTGATCT
1101 CACTGGGGTA GAATTCCCCT GAGAGAATTC CCTCACTCAC GGCTCCCTTT
1151 GCCAGAGTCA GTTCAATCAG GTCTGATGTG AGCAATTTAC ACACTTGTCT
1201 CAGAAAGTCC CTCAGGGTTT GTAGAGGACT GCAGGGGGGC ATCCGCTGCA
1251 GACTCAGCCT TTCTCTGCAG CCATCCTGCA GTGGGGGTGA GCGGGCACAG
1301 GCTGAGAACT GCTCTTGGGT GGTGGAAGCA GGTGTCACGG TGCAAGTCTC
1351 CCCCTGCACC CCTCCCCCAG CTTGAGCCGT GTCACCCCCC TCTCCCTCCA
1401 GCATGGGCCT GTGTCTCAGC TCTCTGGAAG GTGGCCCTGC CCGGACCCTC
1451 TTGCAGGTGT CCTGGTTTGA CTTGGAACTA GATGGCCATC TTTTCAGGCT
1501 TTGGTGGCCC AAGAGCAGTC TGGGTGGATG GAAGTGGCTG TCCCCTTCTC
1551 TTCAGCCCCT GCCCACCCAC TGGTGGAGGT GCTAACTAGC AGGACGTGGC
1601 ATAGGATGGG AGCTGGGCGT GAGGTGCTTT GGGGTCCATT CTTTGTCCCT
1651 CAGCTTCTCA GAGTCCGGCC AGCCCTTGTG TTCCCGTGCC CCACACTTTC
1701 CTCCTCCCCA CTGCAGTGAG TCAATAGTCC AGGGTGGGGC CTGGCCTCCC
1751 TGCCCTGATT GGGGACTCAG GAGGTAAGGC CTGGGGGGCT TCCTGCCCCC
1801 TCCTTGCCCA CCTGCCTGCC CCCGGGCAGC ACGGGAGGGA AAGCAGGGTG
1851 AGCACGCTTG TTGGTTTCAA ATGCACTTTC TGCTTGCATT GCCGTATCTG
1901 TGCGTTCCTT CATCCTGGTC CTGGCTTTAT GGAACACCAT GTTTTTAGCA
1951 TGTTTTTAAA TAAAAACGGA TAAAGTGTCA AAAGCCAAAA AAAAAAAAAA
2001 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA
B: aminoacid sequence (SEQ ID NO:11) length: 157 amino acid
1 MWGRPTFEQS CPALVPWPGF LVRSFSRLRR NPQSADSGAD TSGRRDSADK
51 CCSCTVGPGA SCVFCCGWGG WVGLCLSMQF LIFVNINSKS LVHWEMCNLP
101 ENLFCFWSTS GVASGPRAFA TVLPPAPTSS VCLQSLIYRS PRCLLYSLCA
151 WPFCYLA
C. Nucleotide and amino acid composite sequence (SEQ ID NO:12)
Clone number and protein name: PP1057
Start code: 498 ATG stop coding: 971 TAG
Protein molecular weight: 17224.06
1 GG CAC AGA GCT GGG CAT GGG AGA GAA ACG GGC TGG GGA GTG AGG CCA 47
48 GGA GTG GGA TTC AGC TGC AGC AGG GCG CCC CCT CCA AAC TGC AGC TGG 95
96 TCT GGC TTA CTG TTT TGC CGT TCA AAA AGG TCG CGA ATC CGT GGG ACT 143
144 GAG CAC GGG GAC CTC ACC CGC TAG CCA GCG TCT GCT GCA CTT GAT CAG 191
192 GTG GGG CCT TGG TGG GCG GCT GCC TTT CCT ATA CAG TTT GTC TTG TCA 239
240 CCC TGG TTT CCC ACT GGG GCC AGG TCT CTT CTC CAG CCT CCA CCT GCC 287
288 TGT CTG ATC CAA GAG CTG AGA CAC GGC CAC CCA GCA CCA GTC ACT CCT 335
336 CTG TTC ACC TTA AGT AAC ACA CAA ATC GGG AAC AGG AGG ACA GAA CCG 383
384 TTG GCA TTA TCA GGA TTC GTG TTT TGT GGG GGT GGG AGT GGA GAG TAG 431
432 GGT GGT CTT GTG AGT TGT GCA GGG TGA AGA CCG CTT CCC TGA GAC AGG 479
480 GGC AGT GGT GCT GAT GGA ATG TGG GGG AGG CCC ACA TTT GAG CAA AGC 527
1 Met Trp Gly Arg Pro Thr Phe Glu Gln Ser 10
528 TGC CCT GCC CTT GTC CCC TGG CCT GGC TTC CTG GTA AGG AGT TTC AGC 575
11 Cys Pro Ala Leu Val Pro Trp Pro Gly Phe Leu Val Arg Ser Phe Ser 26
576 CGC CTC CGC AGG AAC CCC CAA AGT GCA GAT TCC GGA GCA GAC ACA TCC 623
27 Arg Leu Arg Arg Asn Pro Gln Ser Ala Asp Ser Gly Ala Asp Thr Ser 42
624 GGG CGG AGA GAC TCA GCA GAC AAG TGC TGC AGT TGC ACG GTG GGA CCC 671
43 Gly Arg Arg Asp Ser Ala Asp Lys Cys Cys Ser Cys Thr Val Gly Pro 58
672 GGG GCC TCG TGC GTT TTT TGC TGT GGG TGG GGT GGG TGG GTT GGT TTA 719
59 Gly Ala Ser Cys Val Phe Cys Cys Gly Trp Gly Gly Trp Val Gly Leu 74
720 TGC CTA TCA ATG CAA TTT TTA ATT TTT GTT AAT ATC AAC AGC AAA AGC 767
75 Cys Leu Ser Met Gln Phe Leu Ile Phe Val Asn Ile Asn Ser Lys Ser 90
768 CTA GTG CAT TGG GAG ATG TGC AAC CTC CCT GAA AAT CTT TTC TGT TTC 815
91 Leu Val His Trp Glu Met Cys Asn Leu Pro Glu Asn Leu Phe Cys Phe 106
816 TGG AGT ACT TCA GGG GTG GCC TCT GGC CCC AGA GCC TTT GCC ACA GTG 863
107 Trp Ser Thr Ser Gly Val Ala Ser Gly Pro Arg Ala Phe Ala Thr Val 122
864 CTC CCA CCA GCC CCC ACC TCA TCC GTC TGT TTG CAG AGC CTC ATC TAC 911
123 Leu Pro Pro Ala Pro Thr Ser Ser Val Cys Leu Gln Ser Leu Ile Tyr 138
912 AGG TCC CCA CGC TGC CTT CTT TAC TCA CTC TGC GCT TGG CCG TTT TGT 959
139 Arg Ser Pro Arg Cys Leu Leu Tyr Ser Leu Cys Ala Trp Pro Phe Cys 154
960 TAT TTG GCT TAG TCT ACA TTG GGC GGA AGT CTG TGT GCA CAG AGT GGG 1007
155 Tyr Leu Ala *** 158
1008 TGT TCC TTC GAG CCC CTT CCA CTC AGA GGG CCA CAC CCA GCG ATG CCA 1055
1056 GTG AAG GTG GCA CAG CCT CTC TTC AGT TTC TCC TGA CTG TGA TCT CAC 1103
1104 TGG GGT AGA ATT CCC CTG AGA GAA TTC CCT CAC TCA CGG CTC CCT TTG 1151
1152 CCA GAG TCA GTT CAA TCA GGT CTG ATG TGA GCA ATT TAC ACA CTT GTC 1199
1200 TCA GAA AGT CCC TCA GGG TTT GTA GAG GAC TGC AGG GGG GCA TCC GCT 1247
1248 GCA GAC TCA GCC TTT CTC TGC AGC CAT CCT GCA GTG GGG GTG AGC GGG 1295
1296 CAC AGG CTG AGA ACT GCT CTT GGG TGG TGG AAG CAG GTG TCA CGG TGC 1343
1344 AAG TCT CCC CCT GCA CCC CTC CCC CAG CTT GAG CCG TGT CAC CCC CCT 1391
1392 CTC CCT CCA GCA TGG GCC TGT GTC TCA GCT CTC TGG AAG GTG GCC CTG 1439
1440 CCC GGA CCC TCT TGC AGG TGT CCT GGT TTG ACT TGG AAC TAG ATG GCC 1487
1488 ATC TTT TCA GGC TTT GGT GGC CCA AGA GCA GTC TGG GTG GAT GGA AGT 1535
1536 GGC TGT CCC CTT CTC TTC AGC CCC TGC CCA CCC ACT GGT GGA GGT GCT 1583
1584 AAC TAG CAG GAC GTG GCA TAG GAT GGG AGC TGG GCG TGA GGT GCT TTG 1631
1632 GGG TCC ATT CTT TGT CCC TCA GCT TCT CAG AGT CCG GCC AGC CCT TGT 1679
1680 GTT CCC GTG CCC CAC ACT TTC CTC CTC CCC ACT GCA GTG AGT CAA TAG 1727
1728 TCC AGG GTG GGG CCT GGC CTC CCT GCC CTG ATT GGG GAC TCA GGA GGT 1775
1776 AAG GCC TGG GGG GCT TCC TGC CCC CTC CTT GCC CAC CTG CCT GCC CCC 1823
1824 GGG CAG CAC GGG AGG GAA AGC AGG GTG AGC ACG CTT GTT GGT TTC AAA 1871
1872 TGC ACT TTC TGC TTG CAT TGC CGT ATC TGT GCG TTC CTT CAT CCT GGT 1919
1920 CCT GGC TTT ATG GAA CAC CAT GTT TTT AGC ATG TTT TTA AAT AAA AAC 1967
1968 GGA TAA AGT GTC AAA AGC CAA AAA AAA AAA AAA AAA AAA AAA AAA AAA 2015
2016 AAA AAA AAA AAA AAA 2030
5.PP1551
A: nucleotide sequence (SEQ ID NO:13) length: 2332bp
1 CACCTCTGGG GAAGGTGTGA GAGGAGAGAA GGAAGTGGCT GTTTGGCTGC
51 TGACAACATG AAGACTTCCT GCGATGAGAA CAGAGGCACA GGTGCCGGCC
101 CTGCAGCCCC CAGAACCCGG ACTGGAGGGG GCCATGGGGC GCCGGACCCT
151 GGCCCTGCCC TGGGTGCTGC TGACCCTGCG TGTCACTGCA GGGACCCCGG
201 AGGTGTGAGT ACAAGTTCGG ATGGAGGCCA CCGAGCTCTC GTCCTTCACC
251 ATCCGTTGTG GGTTCCTGGA GTCTGGCTCC ATCTCCCTGG TGACTGTGAG
301 CAGGGGGGGC CCCGATGGTG CTGGGGGGAC CACGCTGGCT GTGTTGCACC
351 CGGTTCTTGG CATCCAGCAA TGGGCCCCTG CTCGCCAGGC CCGCTGGGAA
401 ACCCAGAGCA GCATCTCTCT CATCCTGGAA GGCTCTGGGG CCAGCAGCCC
451 CTGCGCCAAC ACCACCTTCT GCTGCAAGTT TGCGTCCTTC CCTGAGGGCT
501 CCTGGGAGGC CTGTGGGAGC CTCCCGCCCA GCTCAGACCC AGGTGGGGCC
551 GGGGTGAGGG GTCCAGGAGG GCAGGGAGGG GCTCGGGAAC TGGCCATCCA
601 TCTGATTCTT GCCTCTGTGC CCAGGGCTCT CTGTCCCGCC GACTCCTGCC
651 CCCATTCTGC GGGCAGACCT GGCCGGGATC TTGGGGGTCT CAGGAGTCCT
701 TCTCTTTGAC TGTGGCTACC TCCTTCATCT GCTGTGCCGA CAGAAGCACC
751 GCCCTGCCCC TAGGCTCCAG CCATCCCACA CCAGCTCCTA GGCACTGAGA
801 GCACGAGCAT GGGCATAGCG CGCTGGCTAG GACTCCAGTA CCGTGAAGGG
851 AGGCAGTGAG AGCAGACATC TGTGCCTCAT TCCTGATCTC AAGGGGAAAG
901 CAAGAACAAG ACCTGTGAAG GGAAGGAGGG TGGATCGGCC TGACTGGCCC
951 TGGGAAGCAC AGAGGAGCCT ACTGGGACCA GGGAAGAGAC TGACAAGAGA
1001 AGGGAGGCCT GTCCCTGCCT GCTTCTGTCT CAAGAAGATT GTTATCACTT
1051 CCTCTGGTGA AATCCTGCCT TCCTGTCCCC ACGGCCCCTC AGCTGATGCC
1101 ACCTTACTCT GGAGAGGATG GGCAGAGCCT CCTATGGAGG GAGCCCTGTG
1151 GGTCACCAAA GCTCCAGTGA CCACCCCCGG GCAAAGACCT GCAGATCCCC
1201 TGTTAGAGAC GGGCCCAGGA CTTTGTGCGG GGTGCCCACT GTTGCTCTGA
1251 GCCTTCACTT CCTCAGGGAG GCTTCCTCAG GATCTCGAAC CTGCGGAAGG
1301 AGGACCAGTC TGTGTACTTC TGCCAAGTCC AGCTGGACAT ACAGATCAGG
1351 GAGGCTGTCG TGGCAGTCCA TCAAGGGGAC CCACCTCACC ATCACCCAGG
1401 CCCTCAGGCA GCCCCTCCAC AGGGCCCCTC TCCTGCCTGG ACAGCTCTGC
1451 TGGTCTCCCC GTCCCCTGGA GAAGAACAAG GCCATGGGTC GGCCCCTGCT
1501 GCTGCCCCTG CTGCTCCTGC TGCAGCCGCC AGCATTTCTG CAGCCTGGAT
1551 TATGTGAACC GGCTCTTTCT GAACTGGACA GAGGGTCAGG AGAGCGGCTT
1601 CCTCAGGATC TCAAACCTGC GGAAGGAGGA CCAGTCTGTG TATTTCTGCC
1651 GAGTCGAGCT GGACACCCGG AGATCAGGGA GGCAGCAGTT GCAGTCCATC
1701 AAGGGGACCA AACTCACCAT CACCCAGGCT GTCACAACCA CCACCACCTG
1751 GAGGCCCAGC AGCACAACCA CCATAGCCGG CCTCAGGGTC ACAGAAAGCA
1801 AAGGGCACTC AGAATCATGG CACCTAAGTC TGGACACTGC CATCAGGGTT
1851 GCATTGGCTG TCGCTGTGCT CAAAACTGTC ATTTTGGGAC TGCTGTGCCT
1901 CCTCCTCCTG TGGTGGAGGA GAAGGAAAGG TAGCAGGGCG CCAAGCAGTG
1951 ACTTCTGACC AACAGAGTGT GGGGAGAAGG GATGTGTATT AGCCCCGGAG
2001 GACGTGATGT GAGACCCGCT TGTGAGTCCT CCACACTCGT TCCCCATTGG
2051 CAAGATACAT GGAGAGCACC CTGAGGACCT TTAAAAGGCA AAGCCGCAAG
2101 GCAGAAGGAG GCTGGGTCCC TGAATCACCG ACTGGAGGAG AGTTACCTAC
2151 AAGAGCCTTC ATCCAGGAGC ATCCACACTG CAATGATATA GGAATGAGGT
2201 CTGAACTCCA CTGAATTAAA CCACTGGCAT TTGGGGGCTG TTTATTATAG
2251 CAGTGCAAAG AGTTCCTTTA TCCTCCCCAA GGATGGAAAA ATACAATTTA
2301 TTTTGCTTAC CATAAAAAAA AAAAAAAAAA AA
B: aminoacid sequence (SEQ ID NO:14) length: 271 amino acid
1 MGRASYGGSP VGHQSSSDHP RAKTCRSPVR DGPRTLCGVP TVALSLHFLR
51 EASSGSRTCG RRTSLCTSAK SSWTYRSGRL SWQSIKGTHL TITQALRQPL
101 HRAPLLPGQL CWSPRPLEKN KAMGRPLLLP LLLLLQPPAF LQPGLCEPAL
151 SELDRGSGER LPQDLKPAEG GPVCVFLPSR AGHPEIREAA VAVHQGDQTH
201 HHPGCHNHHH LEAQQHNHHS RPQGHRKQRA LRIMAPKSGH CHQGCIGCRC
251 AQNCHFGTAV PPPPVVEEKE R
C: Nucleotide and amino acid composite sequence (SEQ ID NO:15)
Clone number and protein name: PP1551
Start code: 1118 ATG stop coding: 1933 TAG
Protein molecular weight: 29628.42
1 C ACC TCT GGG GAA GGT GTG AGA GGA GAG AAG GAA GTG GCT GTT TGG 46
47 CTG CTG ACA ACA TGA AGA CTT CCT GCG ATG AGA ACA GAG GCA CAG GTG 94
95 CCG GCC CTC CAG CCC CCA GAA CCC GGA CTG GAG GGG GCC ATG GGG CGC 142
143 CGG ACC CTG GCC CTG CCC TGG GTG CTG CTG ACC CTG CGT GTC ACT GCA 190
191 GGG ACC CCG GAG GTG TGA GTA CAA GTT CGG ATG GAG GCC ACC GAG CTC 238
239 TCG TCC TTC ACC ATC CGT TGT GGG TTC CTG GAG TCT GGC TCC ATC TCC 286
287 CTG GTG ACT GTG AGC AGG GGG GGC CCC GAT GGT GCT GGG GGG ACC ACG 334
335 CTG GCT GTG TTG CAC CCG GAA CTT GGC ATC CAG CAA TGG GCC CCT GCT 382
383 CGC CAG GCC CGC TGG GAA ACC CAG AGC AGC ATC TCT CTC ATC CTG GAA 430
431 GGC TCT GGG GCC AGC AGC CCC TGC GCC AAC ACC ACC TTC TGC TGC AAG 478
479 TTT GCG TCC TTC CCT GAG GGC TCC TGG GAG GCC TGT GGG AGC CTC CCG 526
527 CCC AGC TCA GAC CCA GGT GGG GCC GGG GTG AGG GGT CCA GGA GGG CAG 574
575 GGA GGG GCT CGG GAA CTG GCC ATC CAT CTG ATT CTT GCC TCT GTG CCC 622
623 AGG GCT CTC TGT CCC GCC GAC TCC TGC CCC CAT TCT GCG GGC AGA CCT 670
671 GGC CGG GAT CTT GGG GGT CTC AGG AGT CCT TCT CTT TGA CTG TGG CTA 718
719 CCT CCT TCA TCT GCT GTG CCG ACA GAA GCA CCG CCC TGC CCC TAG GCT 766
767 CCA GCC ATC CCA CAC CAG CTC CTA GGC ACT GAG AGC ACG AGC ATG GGC 814
815 ATA GCG CGC TGG CTA GGA CTC CAG TAC CGT GAA GGG AGG CAG TGA GAG 862
863 CAG ACA TCT GTG CCT CAT TCC TGA TCT CAA GGG GAA AGC AAG AAC AAG 910
911 ACC TGT GAA GGG AAG GAG GGT GGA TCG GCC TGA CTG GCC CTG GGA AGC 958
959 ACA GAG GAG CCT ACT GGG ACC AGG GAA GAG ACT GAC AAG AGA AGG GAG 1006
1007 GCC TGT CCC TGC CTG CTT CTG TCT CAA GAA GAT TGT TAT CAC TTC CTC 1054
1055 TGG TGA AAT CCT GCC TTC CTG TCC CCA CGG CCC CTC AGC TGA TGC CAC 1102
1103 CTT ACT CTG GAG AGG ATG GGC AGA GCC TCC TAT GGA GGG AGC CCT GTG 1150
1 Met Gly Arg Ala Ser Tyr Gly Gly Ser Pro Val 11
1151 GGT CAC CAA AGC TCC AGT GAC CAC CCC CGG GCA AAG ACC TGC AGA TCC 1198
12 Gly His Gln Ser Ser Ser Asp His Pro Arg Ala Lys Thr Cys Arg Ser 27
1199 CCT GTT AGA GAC GGG CCC AGG ACT TTG TGC GGG GTG CCC ACT GTT GCT 1246
28 Pro Val Arg Asp Gly Pro Arg Thr Leu Cys Gly Val Pro Thr Val Ala 43
1247 CTG AGC CTT CAC TTC CTC AGG GAG GCT TCC TCA GGA TCT CGA ACC TGC 1294
44 Leu Ser Leu His Phe Leu Arg Glu Ala Ser Ser Gly Ser Arg Thr Cys 59
1295 GGA AGG AGG ACC AGT CTG TGT ACT TCT GCC AAG TCC AGC TGG ACA TAC 1342
60 Gly Arg Arg Thr Ser Leu Cys Thr Ser Ala Lys Ser Ser Trp Thr Tyr 75
1343 AGA TCA GGG AGG CTG TCG TGG CAG TCC ATC AAG GGG ACC CAC CTC ACC 1390
76 Arg Ser Gly Arg Leu Ser Trp Gln Ser Ile Lys Gly Thr His Leu Thr 91
1391 ATC ACC CAG GCC CTC AGG CAG CCC CTC CAC AGG GCC CCT CTC CTG CCT 1438
92 Ile Thr Gln Ala Leu Arg Gln Pro Leu His Arg Ala Pro Leu Leu Pro 107
1439 GGA CAG CTC TGC TGG TCT CCC CGT CCC CTG GAG AAG AAC AAG GCC ATG 1486
108 Gly Gln Leu Cys Trp Ser Pro Arg Pro Leu Glu Lys Asn Lys Ala Met 123
1487 GGT CGG CCC CTG CTG CTG CCC CTG CTG CTC CTG CTG CAG CCG CCA GCA 1534
124 Gly Arg Pro Leu Leu Leu Pro Leu Leu Leu Leu Leu Gln Pro Pro Ala 139
1535 TTT CTG CAG CCT GGA TTA TGT GAA CCG GCT CTT TCT GAA CTG GAC AGA 1582
140 Phe Leu Gln Pro Gly Leu Cys Glu Pro Ala Leu Ser Glu Leu Asp Arg 155
1583 GGG TCA GGA GAG CGG CTT CCT CAG GAT CTC AAA CCT GCG GAA GGA GGA 1630
156 Gly Ser Gly Glu Arg Leu Pro Gln Asp Leu Lys Pro Ala Glu Gly Gly 171
1631 CCA GTC TGT GTA TTT CTG CCG AGT CGA GCT GGA CAC CCG GAG ATC AGG 1678
172 Pro Val Cys Val Phe Leu Pro Ser Arg Ala Gly His Pro Glu Ile Arg 187
1679 GAG GCA GCA GTT GCA GTC CAT CAA GGG GAC CAA ACT CAC CAT CAC CCA 1726
188 Glu Ala Ala Val Ala Val His Gln Gly Asp Gln Thr His His His Pro 203
1727 GGC TGT CAC AAC CAC CAC CAC CTG GAG GCC CAG CAG CAC AAC CAC CAT 1774
204 Gly Cys His Asn His His His Leu Glu Ala Gln Gln His Asn His His 219
1775 AGC CGG CCT CAG GGT CAC AGA AAG CAA AGG GCA CTC AGA ATC ATG GCA 1822
220 Ser Arg Pro Gln Gly His Arg Lys Gln Arg Ala Leu Arg Ile Met Ala 235
1823 CCT AAG TCT GGA CAC TGC CAT CAG GGT TGC ATT GGC TGT CGC TGT GCT 1870
236 Pro Lys Ser Gly His Cys His Gln Gly Cys Ile Gly Cys Arg Cys Ala 251
1871 CAA AAC TGT CAT TTT GGG ACT GCT GTG CCT CCT CCT CCT GTG GTG GAG 1918
252 Gln Asn Cys His Phe Gly Thr Ala Val Pro Pro Pro Pro Val Val Glu 267
1919 GAG AAG GAA AGG TAG CAG GGC GCC AAG CAG TGA CTT CTG ACC AAC AGA 1966
268 Glu Lys Glu Arg *** 272
1967 GTG TGG GGA GAA GGG ATG TGT ATT AGC CCC GGA GGA CGT GAT GTG AGA 2014
2015 CCC GCT TGT GAG TCC TCC ACA CTC GTT CCC CAT TGG CAA GAT ACA TGG 2062
2063 AGA GCA CCC TGA GGA CCT TTA AAA GGC AAA GCC GCA AGG CAG AAG GAG 2110
2111 GCT GGG TCC CTG AAT CAC CGA CTG GAG GAG AGT TAC CTA CAA GAG CCT 2158
2159 TCA TCC AGG AGC ATC CAC ACT GCA ATG ATA TAG GAA TGA GGT CTG AAC 2206
2207 TCC ACT GAA TTA AAC CAC TGG CAT TTG GGG GCT GTT TAT TAT AGC AGT 2254
2255 GCA AAG AGT TCC TTT ATC CTC CCC AAG GAT GGA AAA ATA CAA TTT ATT 2302
2303 TTG CTT ACC ATA AAA AAA AAA AAA AAA AAA 2332
6.PP1553
A: nucleotide sequence (SEQ ID NO:16) length: 2265bp
1 GGAAGGAGGA GGTCCAGGCC GGTAGTGGCC ATTCGCCGTG CCTCAGCGAG
51 CAGGTGTGTG TGGGTCCTCC ACCACTCACC TCTTGGTTAG CGGGAGTGTG
101 CTGCCCCCAC CCCCACCCCC GCACCCCCAT TCTACACAAG GCAGAAGAGG
151 CACGGGTTTT TCTGGGAGCG AATATCAAGT GCCTGAGAGC AACTACAGGA
201 CTAACTGTGT TTGGGTTGGG TGTAGTATAA ATAATAATAA TGGCTAATAT
251 TTCCTGAGCA TCTACTAAAT GCAAGGAATT GTGCTTGGTG TGTCATGTGG
301 ATTCTCTCTT GCATCTTCAT GATAAATGTT ATTGTCGCTG TTTTACCGAT
351 GAGGGTTGGA TTAGAGGGGT TAAACAACTT GTCTTAGGCT CCACAGCTGG
401 GAACAAGTGG GGCTGGGAAG CTGACTTCGT GCTCTTCACC ACCACAAAGG
451 ATGTGTGTGC ATCCTGGGGC ATGCCTGCCT CATGTGGGGG TGTCCTGGGC
501 TGAATTTCCT GGGCACTTCT CAGTGGAACT CTCTAGCCTC CTGGTTCGGA
551 ATGTCAACTA TGAGATCCCC TCACTGAAGA AGCAGATTGC CAAGTGCCAG
601 CAGCTGCAGC AAGAATACAG CCGCAAGGAG GAGGAGTGCC AGGCAGGGGC
651 TGCCGAGATG CGGGAGCAGT TCTACCACTC CTGCAAGCAG TATGGCATCA
701 CGGGCGAAAA TGTCCGAGGA GAACTGCTGG CCCTGGTGAA GGACCTGCCG
751 AGTCAGCTGG CTGAGATTGG GGCAGCGGCT CAGCAGTCCC TGGGGGAAGC
801 CATTGACGTG TACCAGGCGT CTGTGGGGTT TGTGTGTGAG AGGTAGAGAG
851 GCCTCAGCTT CTCCTGGTGG GGGTGCTTTG CCTGTGTTCC CCAGCTCATG
901 ACCCTTCTCC AGTTGTCTTG TTCCCATATA ACATTTGAAC TCTTTACACA
951 CCTGAACCTG TGGGGGCCTT GCCCATTTGA CCATGTGGCC CAGGCCAAAG
1001 CCCAGTGTTG GCCTTACGCA TGGTCGGCAG GAGAGTCAGT TGTGTGCTCT
1051 GTTGAAGCCC CACAGAGCAG GTGTTGCCAA TGCTGCGGTT CGTGCAGAAG
1101 CGGGGAAACT CAACGGTGTA CGAGTGGAGG ACAGGGACAG AGCCCTCTGT
1151 GGTGGAACGA CCCCACCTCG AGGAGCTTCC TGAGCAGGTG GCAGAAGATG
1201 CGATTGACTG GGGCGACTTT GGGGTAGAGG CAGTGTCTGA GGGGACTGAC
1251 TCTGGCATCT CTGCCGAGGC TGCTGGAATC GACTGGGGCA TCTTCCCGGA
1301 ATCAGATTCA AAGGATCCTG GAGGTGATGG GATAGACTGG GGAGACGATG
1351 CTGTTGCTTT GCAGATCACA GTGCTGGAAG CAGGAACCCA GGCTCCAGAA
1401 GGTGTTGCCA GGGGCCCAGA TGCCCTGACA CTGCTTGAAT ACACTGAGAC
1451 CCGGAATCAG TTCCTTGATG AGCTCATGGA GCTTGAGATC TTCTTAGCCC
1501 AGAGAGCAGT GGAGTTGAGT GAGGAGGCAG ATGTCCTGTC TGTGAGCCAG
1551 TTCCAGCTGG CTCCAGCCAT CCTGCAGGGC CAGACCAAAG AGAAGATGGT
1601 TACCATGGTG TCAGTGCTGG AGGATCTGAT TGGCAAGCTT ACCAGTCTTC
1651 AGCTGCAACA CCTGTTTATG ATCCTGGCCT CACCAAGGTA TGTGGACCGA
1701 GTGACTGAAT TCCTCCAGCA AAAGCTGAAG CAGTCCCAGC TGCTGGCTTT
1751 GAAGAAAGAG CTGATGGTGC AGAAGCAGCA GGAGGCACTT GAGGAGCAGG
1801 CGGCTCTGGA GCCTAAGCTG GACCTGCTAC TGGAGAAGAC CAAGGAGCTG
1851 CAGAAGCTGA TTGAAGCTGA CATCTCCAAG AGGTACAGCG GGCGCCCTGT
1901 GAACCTGATG GGAACCTCTC TGTGACACCC TCCGTGTTCT TGCCTGCCCA
1951 TCTTCTCCGC TTTTGGGATG AAGATGATAG CCAGGGCTGT TGTTTTGGGG
2001 CCCTTCAAGG CAAAAGACCA GGCTGACTGG AAGATGGAAA GCCACAGGAA
2051 GGAAGCGGCA CCTGATGGTG ATCTTGGCAC TCTCCATGTT CTCTACAAGA
2101 AGCTGTGGTG ATTGGCCCTG TGGTCTATCA GGCGAAAACC ACAGATTCTC
2151 CTTCTAGTTA GTATAGCGGA CTTAATAAAA GAGGAAAAAA CTCTTTAAAA
2201 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA
2251 AAAAAAAAAA AAAAA
B: aminoacid sequence (SEQ ID NO:17) length: 281 amino acid
1 MLRFVQKRGN STVYEWRTGT EPSVVERPHL EELPEQVAED AIDWGDFGVE
51 AVSEGTDSGI SAEAAGIDWG IFPESDSKDP GGDGIDWGDD AVALQITVLE
101 AGTQAPEGVA RGPDALTLLE YTETRNQFLD ELMELEIFLA QRAVELSEEA
151 DVLSVSQFQL APAILQGQTK EKMVTMVSVL EDLIGKLTSL QLQHLFMILA
201 SPRYVDRVTE FLQQKLKQSQ LLALKKELMV QKQQEALEEQ AALEPKLDLL
251 LEKTKELQKL IEADISKRYS GRPVNLMGTS L
C: Nucleotide and aminoacid sequence (SEQ ID NO:18)
Clone number and protein name: PP1553
Start code: 1080 ATG stop coding: 1925 TGA
Protein molecular weight: 31198.70
1 GG AAG GAG GAG GTC CAG GCC GGT AGT GGC CAT TCG CCG TGC CTC AGC 47
48 GAG CAG GTG TGT GTG GGT CCT CCA CCA CTC ACC TCT TGG TTA GCG GGA 95
96 GTG TGC TGC CCC CAC CCC CAC CCC CGC ACC CCC ATT CTA CAC AAG GCA 143
144 GAA GAG GCA CGG GTT TTT CTG GGA GCG AAT ATC AAG TGC CTG AGA GCA 191
192 ACT ACA GGA CTA ACT GTG TTT GGG TTG GGT GTA GTA TAA ATA ATA ATA 239
240 ATG GCT AAT ATT TCC TGA GCA TCT ACT AAA TGC AAG GAA TTG TGC TTG 287
288 GTG TGT CAT GTG GAT TCT CTC TTG CAT CTT CAT GAT AAA TGT TAT TGT 335
336 CGC TGT TTT ACC GAT GAG GGT TGG ATT AGA GGG GTT AAA CAA CTT GTC 383
384 TTA GGC TCC ACA GCT GGG AAC AAG TGG GGC TGG GAA GCT GAC TTC GTG 431
432 CTC TTC ACC ACC ACA AAG GAT GTG TGT GCA TCC TGG GGC ATG CCT GCC 479
480 TCA TGT GGG GGT GTC CTG GGC TGA ATT TCC TGG GCA CTT CTC AGT GGA 527
528 ACT CTC TAG CCT CCT GGT TCG GAA TGT CAA CTA TGA GAT CCC CTC ACT 575
576 GAA GAA GCA GAT TGC CAA GTG CCA GCA GCT GCA GCA AGA ATA CAG CCG 623
624 CAA GGA GGA GGA GTG CCA GGC AGG GGC TGC CGA GAT GCG GGA GCA GTT 671
672 CTA CCA CTC CTG CAA GCA GTA TGG CAT CAC GGG CGA AAA TGT CCG AGG 719
720 AGA ACT GCT GGC CCT GGT GAA GGA CCT GCC GAG TCA GCT GGC TGA GAT 767
768 TGG GGC AGC GGC TCA GCA GTC CCT GGG GGA AGC CAT TGA CGT GTA CCA 815
816 GGC GTC TGT GGG GTT TGT GTG TGA GAG GTA GAG AGG CCT CAG CTT CTC 863
864 CTG GTG GGG GTG CTT TGC CTG TGT TCC CCA GCT CAT GAC CCT TCT CCA 911
912 GTT GTC TTG TTC CCA TAT AAC ATT TGA ACT CTT TAC ACA CCT GAA CCT 959
960 GTG GGG GCC TTG CCC ATT TGA CCA TGT GGC CCA GGC CAA AGC CCA GTG 1007
1008 TTG GCC TTA CGC ATG GTC GGC AGG AGA GTC AGT TGT GTG CTC TGT TGA 1055
1056 AGC CCC ACA GAG CAG GTG TTG CCA ATG CTG CGG TTC GTG CAG AAG CGG 1103
1 Met Leu Arg Phe Val Gln Lys Arg 8
1104 GGA AAC TCA ACG GTG TAC GAG TGG AGG ACA GGG ACA GAG CCC TCT GTG 1151
9 Gly Asn Ser Thr Val Tyr Glu Trp Arg Thr Gly Thr Glu Pro Ser Val 24
1152 GTG GAA CGA CCC CAC CTC GAG GAG CTT CCT GAG CAG GTG GCA GAA GAT 1199
25 Val Glu Arg Pro His Leu Glu Glu Leu Pro Glu Gln Val Ala Glu Asp 40
1200 GCG ATT GAC TGG GGC GAC TTT GGG GTA GAG GCA GTG TCT GAG GGG ACT 1247
41 Ala Ile Asp Trp Gly Asp Phe Gly Val Glu Ala Val Ser Glu Gly Thr 56
1248 GAC TCT GGC ATC TCT GCC GAG GCT GCT GGA ATC GAC TGG GGC ATC TTC 1295
57 Asp Ser Gly Ile Ser Ala Glu Ala Ala Gly Ile Asp Trp Gly Ile Phe 72
1296 CCG GAA TCA GAT TCA AAG GAT CCT GGA GGT GAT GGG ATA GAC TGG GGA 1343
73 Pro Glu Ser Asp Ser Lys Asp Pro Gly Gly Asp Gly Ile Asp Trp Gly 88
1344 GAC GAT GCT GTT GCT TTG CAG ATC ACA GTG CTG GAA GCA GGA ACC CAG 1391
89 Asp Asp Ala Val Ala Leu Gln Ile Thr Val Leu Glu Ala Gly Thr Gln 104
1392 GCT CCA GAA GGT GTT GCC AGG GGC CCA GAT GCC CTG ACA CTG CTT GAA 1439
105 Ala Pro Glu Gly Val Ala Arg Gly Pro Asp Ala Leu Thr Leu Leu Glu 120
1440 TAC ACT GAG ACC CGG AAT CAG TTC CTT GAT GAG CTC ATG GAG CTT GAG 1487
121 Tyr Thr Glu Thr Arg Asn Gln Phe Leu Asp Glu Leu Met Glu Leu Glu 136
1488 ATC TTC TTA GCC CAG AGA GCA GTG GAG TTG AGT GAG GAG GCA GAT GTC 1535
137 Ile Phe Leu Ala Gln Arg Ala Val Glu Leu Ser Glu Glu Ala Asp Val 152
1536 CTG TCT GTG AGC CAG TTC CAG CTG GCT CCA GCC ATC CTG CAG GGC CAG 1583
153 Leu Ser Val Ser Gln Phe Gln Leu Ala Pro Ala Ile Leu Gln Gly Gln 168
1584 ACC AAA GAG AAG ATG GTT ACC ATG GTG TCA GTG CTG GAG GAT CTG ATT 1631
169 Thr Lys Glu Lys Met Val Thr Met Val Ser Val Leu Glu Asp Leu Ile 184
1632 GGC AAG CTT ACC AGT CTT CAG CTG CAA CAC CTG TTT ATG ATC CTG GCC 1679
185 Gly Lys Leu Thr Ser Leu Gln Leu Gln His Leu Phe Met Ile Leu Ala 200
1680 TCA CCA AGG TAT GTG GAC CGA GTG ACT GAA TTC CTC CAG CAA AAG CTG 1727
201 Ser Pro Arg Tyr Val Asp Arg Val Thr Glu Phe Leu Gln Gln Lys Leu 216
1728 AAG CAG TCC CAG CTG CTG GCT TTG AAG AAA GAG CTG ATG GTG CAG AAG 1775
217 Lys Gln Ser Gln Leu Leu Ala Leu Lys Lys Glu Leu Met Val Gln Lys 232
1776 CAG CAG GAG GCA CTT GAG GAG CAG GCG GCT CTG GAG CCT AAG CTG GAC 1823
233 Gln Gln Glu Ala Leu Glu Glu Gln Ala Ala Leu Glu Pro Lys Leu Asp 248
1824 CTG CTA CTG GAG AAG ACC AAG GAG CTG CAG AAG CTG ATT GAA GCT GAC 1871
249 Leu Leu Leu Glu Lys Thr Lys Glu Leu Gln Lys Leu Ile Glu Ala Asp 264
1872 ATC TCC AAG AGG TAC AGC GGG CGC CCT GTG AAC CTG ATG GGA ACC TCT 1919
265 Ile Ser Lys Arg Tyr Ser Gly Arg Pro Val Asn Leu Met Gly Thr Ser 280
1920 CTG TGA CAC CCT CCG TGT TCT TGC CTG CCC ATC TTC TCC GCT TTT GGG 1967
281 Leu *** 282
1968 ATG AAG ATG ATA GCC AGG GCT GTT GTT TTG GGG CCC TTC AAG GCA AAA 2015
2016 GAC CAG GCT GAC TGG AAG ATG GAA AGC CAC AGG AAG GAA GCG GCA CCT 2063
2064 GAT GGT GAT CTT GGC ACT CTC CAT GTT CTC TAC AAG AAG CTG TGG TGA 2111
2112 TTG GCC CTG TGG TCT ATC AGG CGA AAA CCA CAG ATT CTC CTT CTA GTT 2159
2160 AGT ATA GCG GAC TTA ATA AAA GAG GAA AAA ACT CTT TAA AAA AAA AAA 2207
2208 AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA 2255
2256 AAA AAA AAA A 2265
7.PP1665
A: nucleotide sequence (SEQ ID NO:19) length: 1559bp
1 CTGCCTAGCA GGCAGAGGCC CCTGGTGCGG AAGGTGGCTC CCGGCTTCCA
51 ACAGACATCA GGCTCCAAGG AGGCAGTCGC CAGCCTGCGG AGAGGCCACA
101 TCCAGCGGCT GAACCTGCGC TACACTCAGG TGTCCCGCCA GGAGCTCAGG
151 GACTACGCGT CCTGGAACCT GAGTGTGAAC CTCTACACAG TCAACGCACC
201 GTGGCTCTTC TCCCTGCTGT GGTGTGCGGG GGTCCCATCC GTCACCTCTG
251 ACAACTCCCA CACCCTGTCC CAGGTGCCTT CCCCCCTCTG GATCATGCCC
301 CCGGACGAGT ACTGTCTCAT GTGGGTCACT GCCGACCTGG TCTCCTTCAC
351 CCTCATCGTG GGCATCTTCG TGCTCCAGAA GTGGCGCCTG GGTGGCATAC
401 GGAGCTACAA CCCTGAGCAG ATCATGCTGA GTGCTGCGGT GCGCCGGACC
451 AGCCGGGACG TCAGCATCAT GAAGGAGAAG CTTATTTTCT CAGAGATCAG
501 CGATGGTGTA GAGGTCTCCG ATGTGCTCTC CGTATGTTCA GACAACAGTT
551 ATGACACATA TGCCAACAGC ACCGCCACCC CTGTGGGCCC CCGAGGGGGT
601 GGCAGCCACA CCAAGACCCT CATAGAGCGG AGTGGGCGTT AGCTGAAGAC
651 ATGTCTGTCC CACCTGTACC TGACACAGAA GCTGGGGAGC CTAGGAGAGC
701 TGGTGGAAGT GTGTCTGAAC TCGGAGTGCT CTGGGAGCGG GCTCCACAGC
751 CTCCTTGTGG GCTCCAGCCC CTTGTCAGCC GCAGCCTCTC TTGAGGGGGA
801 CTCCCTGTCT CCTGAGGCCC AGCTGGGCCA GGACTCCATC CTTTCAGATG
851 CCCCTGCAGG CCTGGGGCTC CTTCTGGGAA GTATGGGGCC TAGGGCTTGG
901 TCCCCCTCTT CTGAGGCCCT CTCCTGTATC CCGACCTGGA AGCTTTGATG
951 GGTCATGGGC CATGCCATAC CCCCTGTGGC AATGGAGTGT GTGGATGCTC
1001 ACCTGTGCCA TCTGTCCTCC TGTCTGTGCC AGGAGGCACC TGAGTTCTCT
1051 GCTGTTATCC TGCCCCAAGG GCCTGGGCCG AGCCTCTACC TGAAGCAACT
1101 CTGCTCTTCC TGTCAGTCTC AAAGCACAAG GAGGTTCAGC CCAGGAGGAA
1151 GCCAGCTGCA ATGTGGAGAC ACGTCCTCCT CCCCAACCCA CCTCATGCCA
1201 CCGCCAACCC CCTGCCCCAG GAGCGGGCCT GAGCCACGTC CCCTAGGAGC
1251 AGCTGGAGAT GGCCAAAAGA GTGAGCTCAG GACTACTGGA TCCCATGCCC
1301 AGGTGTCCAG CAGACCTCAA GGCAGAAGGG TCACCTAACC CAGGAGTCCA
1351 CAGACTGATG TGACCTCAGG TTCCCACATC AGTGGCCACA GGGCAGGGCC
1401 CACCTGGTAG AAGTGTTCTG GATATGGCCA GGGTGGGTGT GTGGCTAAGT
1451 GGGCCTGAAC AGAGGGAACC TAGGGCCCTT GGCCAATGTG ATTAAAGCTG
1501 CATCTTGAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA
1551 AAAAAAAAC
B: aminoacid sequence (SEQ ID NO:20) length: 127 amino acid
1 MGHAIPPVAM ECVDAHLCHL SSCLCQEAPE FSAVILPQGP GPSLYLKQLC
51 SSCQSQSTRR FSPGGSQLQC GDTSSSPTHL MPPPTPCPRS GPEPRPLGAA
101 GDGQKSELRT TGSHAQVSSR PQGRRVT
C. Nucleotide and amino acid composite sequence (SEQ ID NO:21)
Clone number and protein name: PP1665
Start code: 955 ATG stop coding: 1338 TAA
Protein molecular weight: 13303.38
1 CTG CCT AGC AGG CAG AGG CCC CTG GTG CGG AAG GTG GCT CCC GGC TTC 48
49 CAA CAG ACA TCA GGC TCC AAG GAG GCA GTC GCC AGC CTG CGG AGA GGC 96
97 CAC ATC CAG CGG CTG AAC CTG CGC TAC ACT CAG GTG TCC CGC CAG GAG 144
145 CTC AGG GAC TAC GCG TCC TGG AAC CTG AGT GTG AAC CTC TAC ACA GTC 192
193 AAC GCA CCG TGG CTC TTC TCC CTG CTG TGG TGT GCG GGG GTC CCA TCC 240
241 GTC ACC TCT GAC AAC TCC CAC ACC CTG TCC CAG GTG CCT TCC CCC CTC 288
289 TGG ATC ATG CCC CCG GAC GAG TAC TGT CTC ATG TGG GTC ACT GCC GAC 336
337 CTG GTC TCC TTC ACC CTC ATC GTG GGC ATC TTC GTG CTC CAG AAG TGG 384
385 CGC CTG GGT GGC ATA CGG AGC TAC AAC CCT GAG CAG ATC ATG CTG AGT 432
433 GCT GCG GTG CGC CGG ACC AGC CGG GAC GTC AGC ATC ATG AAG GAG AAG 480
481 CTT ATT TTC TCA GAG ATC AGC GAT GGT GTA GAG GTC TCC GAT GTG CTC 528
529 TCC GTA TGT TCA GAC AAC AGT TAT GAC ACA TAT GCC AAC AGC ACC GCC 576
577 ACC CCT GTG GGC CCC CGA GGG GGT GGC AGC CAC ACC AAG ACC CTC ATA 624
625 GAG CGG AGT GGG CGT TAG CTG AAG ACA TGT CTG TCC CAC CTG TAC CTG 672
673 ACA CAG AAG CTG GGG AGC CTA GGA GAG CTG GTG GAA GTG TGT CTG AAC 720
721 TCG GAG TGC TCT GGG AGC GGG CTC CAC AGC CTC CTT GTG GGC TCC AGC 768
769 CCC TTG TCA GCC GCA GCC TCT CTT GAG GGG GAC TCC CTG TCT CCT GAG 816
817 GCC CAG CTG GGC CAG GAC TCC ATC CTT TCA GAT GCC CCT GCA GGC CTG 864
865 GGG CTC CTT CTG GGA AGT ATG GGG CCT AGG GCT TGG TCC CCC TCT TCT 912
913 GAG GCC CTC TCC TGT ATC CCG ACC TGG AAG CTT TGA TGG GTC ATG GGC 960
1 Met Gly 2
961 CAT GCC ATA CCC CCT GTG GCA ATG GAG TGT GTG GAT GCT CAC CTG TGC 1008
3 His Ala Ile Pro Pro Val Ala Met Glu Cys Val Asp Ala His Leu Cys 18
1009 CAT CTG TCC TCC TGT CTG TGC CAG GAG GCA CCT GAG TTC TCT GCT GTT 1056
19 His Leu Ser Ser Cys Leu Cys Gln Glu Ala Pro Glu Phe Ser Ala Val 34
1057 ATC CTG CCC CAA GGG CCT GGG CCG AGC CTC TAC CTG AAG CAA CTC TGC 1104
35 Ile Leu Pro Gln Gly Pro Gly Pro Ser Leu Tyr Leu Lys Gln Leu Cys 50
1105 TCT TCC TGT CAG TCT CAA AGC ACA AGG AGG TTC AGC CCA GGA GGA AGC 1152
51 Ser Ser Cys Gln Ser Gln Ser Thr Arg Arg Phe Ser Pro Gly Gly Ser 66
1153 CAG CTG CAA TGT GGA GAC ACG TCC TCC TCC CCA ACC CAC CTC ATG CCA 1200
67 Gln Leu Gln Cys Gly Asp Thr Ser Ser Ser Pro Thr His Leu Met Pro 82
1201 CCG CCA ACC CCC TGC CCC AGG AGC GGG CCT GAG CCA CGT CCC CTA GGA 1248
83 Pro Pro Thr Pro Cys Pro Arg Ser Gly Pro Glu Pro Arg Pro Leu Gly 98
1249 GCA GCT GGA GAT GGC CAA AAG AGT GAG CTC AGG ACT ACT GGA TCC CAT 1296
99 Ala Ala Gly Asp Gly Gln Lys Ser Glu Leu Arg Thr Thr Gly Ser His 114
1297 GCC CAG GTG TCC AGC AGA CCT CAA GGC AGA AGG GTC ACC TAA CCC AGG 1344
115 Ala Gln Val Ser Ser Arg Pro Gln Gly Arg Arg Val Thr *** 128
1345 AGT CCA CAG ACT GAT GTG ACC TCA GGT TCC CAC ATC AGT GGC CAC AGG 1392
1393 GCA GGG CCC ACC TGG TAG AAG TGT TCT GGA TAT GGC CAG GGT GGG TGT 1440
1441 GTG GCT AAG TGG GCC TGA ACA GAG GGA ACC TAG GGC CCT TGG CCA ATG 1488
1489 TGA TTA AAG CTG CAT CTT GAA AAA AAA AAA AAA AAA AAA AAA AAA AAA 1536
1537 AAA AAA AAA AAA AAA AAA AAA AC 1559
8.PP2135
A: nucleotide sequence (SEQ ID NO:22) length: 1188bp
1 GAAAACCCCG GCTTTGAAGC CTCACCACCT GCCCAGGGGA TACCCGAGGC
51 CAAAGTCAGG CACCCCCTGT CCTATGTGGC CCAGCGGCAG CCTTCTGAGT
101 CTGGGCGGCA TCTGCTTTCG GAGCCCAGCA CCCCCCTGTC TCCTCCAGGC
151 CCCGGAGACG TCTTCTTCCC ATCCCTGGAC CCTGTCCCTG ACTCTCCAAA
201 CTTTGAGGTC ATCTAGCCCA GCTGGGGGAC AGTGGGCTGT TGTGGCTGGG
251 TCTGGGGCAG GTGCATTTGA GCCAGGGCTG GCTCTGTGAG TGGCCTCCTT
301 GGCCTCGGCC CTGGTTCCCT CCCTCCTGCT CTGGGCTCAG ATACTGTGAC
351 ATCCCAGAAG CCCAGCCCCT CAACCCCTCT GGATGCTACA TGGGGATGCT
401 GGACGGCTCA GCCCCTGTTC CAAGGATTTT GGGGTGCTGA GATTCTCCCC
451 TAGAGACCTG AAATTCACCA GCTACAGATG CCAAATGACT TACATCTTAA
501 GAAGTCTCAG AACGTCCAGC CCTTCAGCAG CTCTCGTTCT GAGACATGAG
551 CCTTGGGATG TGGCAGCATC AGTGGGACAA GATGGACACT GGGCCACCCT
601 CCCAGGCACC AGACACAGGG CACGGTGGAG AGACTTCTCC CCCGTGGCCG
651 CCTTGGCTCC CCCGTTTTGC CCGAGGCTGC TCTTCTGTCA GACTTCCTCT
701 TTGTACCACA GTGGCTCTGG GGCCAGGCCT GCCTGCCCAC TGGCCATCGC
751 CACCTTCCCC AGCTGCCTCC TACCAGCAGT TTCTCTGAAG ATCTGTCAAC
801 AGGTTAAGTC AATCTGGGGC TTCCACTGCC TGCATTCCAG TCCCCAGAGC
851 TTGGTGGTCC CGAAACGGGA AGTACATATT GGGGCATGGT GGCCTCCGTG
901 AGCAAATGGT GTCTTGGGCA ATCTGAGGCC AGGACAGATG TTGCCCCACC
951 CACTGGAGAT GGTGCTGAGG GAGGTGGGTG GGGCCTTCTG GGAAGGTGAG
1001 TGGAAGAAGG CACCTGCCCC CCGCCCTCCC CATCCCCTAC TCCCACTGCT
1051 CAGCGCGGGC CATTGCAAGG GTGCCACACA ATGTCTTGTC CACCCTGGGA
1101 CACTTCTGAG TATGAAGCGG GATGCTATTA AAAACTACAT GGGGAAACAG
1151 GTGCAAACCC TGGAAAAAAA AAAAAAAAAA AAAAAAA
B: aminoacid sequence (SEQ ID NO:23) length: 172 amino acid
1 MLHGDAGRLS PCSKDFGVLR FSPRDLKFTS YRCQMTYILR SLRTSSPSAA
51 LVLRHEPWDV AASVGQDGHW ATLPGTRHRA RWRDFSPVAA LAPPFCPRLL
101 FCQTSSLYHS GSGARPACPL AIATFPSCLL PAVSLKICQQ VKSIWGFHCL
151 HSSPQSLVVP KREVHIGAWW PP
C: Nucleotide and amino acid composite sequence (SEQ ID NO:24)
Clone number and protein name: PP2135
Start code: 383 ATG stop coding: 901 TGA
Protein molecular weight: 19046.05
1 G AAA ACC CCG GCT TTG AAG CCT CAC CAC CTG CCC AGG GGA TAC CCG 46
47 AGG CCA AAG TCA GGC ACC CCC TGT CCT ATG TGG CCC AGC GGC AGC CTT 94
95 CTG AGT CTG GGC GGC ATC TGC TTT CGG AGC CCA GCA CCC CCC TGT CTC 142
143 CTC CAG GCC CCG GAG ACG TCT TCT TCC CAT CCC TGG ACC CTG TCC CTG 190
191 ACT CTC CAA ACT TTG AGG TCA TCT AGC CCA GCT GGG GGA CAG TGG GCT 238
239 GTT GTG GCT GGG TCT GGG GCA GGT GCA TTT GAG CCA GGG CTG GCT CTG 286
287 TGA GTG GCC TCC TTG GCC TCG GCC CTG GTT CCC TCC CTC CTG CTC TGG 334
335 GCT CAG ATA CTG TGA CAT CCC AGA AGC CCA GCC CCT CAA CCC CTC TGG 382
383 ATG CTA CAT GGG GAT GCT GGA CGG CTC AGC CCC TGT TCC AAG GAT TTT 430
1 Met Leu His Gly Asp Ala Gly Arg Leu Ser Pro Cys Ser Lys Asp Phe 16
431 GGG GTG CTG AGA TTC TCC CCT AGA GAC CTG AAA TTC ACC AGC TAC AGA 478
17 Gly Val Leu Arg Phe Ser Pro Arg Asp Leu Lys Phe Thr Ser Tyr Arg 32
479 TGC CAA ATG ACT TAC ATC TTA AGA AGT CTC AGA ACG TCC AGC CCT TCA 526
33 Cys Gln Met Thr Tyr Ile Leu Arg Ser Leu Arg Thr Ser Ser Pro Ser 48
527 GCA GCT CTC GTT CTG AGA CAT GAG CCT TGG GAT GTG GCA GCA TCA GTG 574
49 Ala Ala Leu Val Leu Arg His Glu Pro Trp Asp Val Ala Ala Ser Val 64
575 GGA CAA GAT GGA CAC TGG GCC ACC CTC CCA GGC ACC AGA CAC AGG GCA 622
65 Gly Gln Asp Gly His Trp Ala Thr Leu Pro Gly Thr Arg His Arg Ala 80
623 CGG TGG AGA GAC TTC TCC CCC GTG GCC GCC TTG GCT CCC CCG TTT TGC 670
81 Arg Trp Arg Asp Phe Ser Pro Val Ala Ala Leu Ala Pro Pro Phe Cys 96
671 CCG AGG CTG CTC TTC TGT CAG ACT TCC TCT TTG TAC CAC AGT GGC TCT 718
97 Pro Arg Leu Leu Phe Cys Gln Thr Ser Ser Leu Tyr His Ser Gly Ser 112
719 GGG GCC AGG CCT GCC TGC CCA CTG GCC ATC GCC ACC TTC CCC AGC TGC 766
113 Gly Ala Arg Pro Ala Cys Pro Leu Ala Ile Ala Thr Phe Pro Ser Cys 128
767 CTC CTA CCA GCA GTT TCT CTG AAG ATC TGT CAA CAG GTT AAG TCA ATC 814
129 Leu Leu Pro Ala Val Ser Leu Lys Ile Cys Gln Gln Val Lys Ser Ile 144
815 TGG GGC TTC CAC TGC CTG CAT TCC AGT CCC CAG AGC TTG GTG GTC CCG 862
145 Trp Gly Phe His Cys Leu His Ser Ser Pro Gln Ser Leu Val Val Pro 160
863 AAA CGG GAA GTA CAT ATT GGG GCA TGG TGG CCT CCG TGA GCA AAT GGT 910
161 Lys Arg Glu Val His Ile Gly Ala Trp Trp Pro Pro *** 173
911 GTC TTG GGC AAT CTG AGG CCA GGA CAG ATG TTG CCC CAC CCA CTG GAG 958
959 ATG GTG CTG AGG GAG GTG GGT GGG GCC TTC TGG GAA GGT GAG TGG AAG 1006
1007 AAG GCA CCT GCC CCC CGC CCT CCC CAT CCC CTA CTC CCA CTG CTC AGC 1054
1055 GCG GGC CAT IGC AAG GGT GCC ACA CAA TGT CTT GTC CAC CCT GGG ACA 1102
1103 CTT CTG AGT ATG AAG CGG GAT GCT ATT AAA AAC TAC ATG GGG AAA CAG 1150
1151 GTG CAA ACC CTG GAA AAA AAA AAA AAA AAA AAA AAA AA 1188
9.PP2281
A: nucleotide sequence (SEQ ID NO:25) length: 1939bp
1 CAGACATTCA CTCTGGCTGC TGGGACATCA GAAAACAAAG TCTTCATCTC
51 TCTCTCCAGT TTCACCCACC CCACCCTTTG CTTTCATTTC AGGTGTGTTG
101 GTCTATATGA CAGGGAGGAG AGTAAAGGAG AGCAGGAGCA ATTGGCTGCC
151 TGCAAAGCCA GCTGGAGGTG AAGTGCAGGA AAGGAAAGGT CACCCCATTC
201 TACTCCATGG CCTCTCTGCT CCCAGCTGTG GTAGGCTCAC ATAGCCAGTG
251 TGATCGGTTT TTAAGAGGCA GTGCTTTTCA GCTTTTCTCC CTGATATATC
301 CATTTTGCTT CCCAGCACTT TTTAGGAGTA GTGAGAGCGC TTCCTGCCCT
351 TGTTGGAAGC CCCAGGGTGG ACTCTCAGCA CGAAGGTCTC TCCCTTAACT
401 GCTGCCCTTC CAAGACTTGC TCCCGAGATG GAGTGGGCGT GGTCTTCCAG
451 GCTGGCCCTT CCTTCTCCTC ACCGCCACCT TCCCTGCCCC AGCCCCAGCA
501 GCCATGGGTA CATGGGTCCC CAGCTCACCT ATGGATTCCC GCCAGTCTGC
551 CCAGCTGCAG TACTCACGCC CCATGGGGGA TCTTGGTCTG TTTTTCTTGT
601 GGGAGCCTAG TGGAGAGCAG ACGTGGCTTT TTATGTGTCT TGTTGGGGAG
651 GTGACTTGCA TGGTGGGGAC AAGGCTGTCG TGGCAACCTT GGGATCGAGT
701 TTGAGACTAA AGGATGTCAT GAGATCCCTG GCTTCTCCCC ATGTTGTTCC
751 CGGACAAGGG CAGAAGGGAG GCATGGCAAG GGACCTCTGC TGTCCTTACT
801 CAACAGTGGT CCTCATCCCT CCCCACCTCC CACTGCTTCC TGCAAGGGCA
851 CCAGTTGTAT GAGAAAGTTG GCCTTTGGAC TTAGGATTTC TTATTGTAGC
901 TAAGAGCCAT CTGAAGCAGC AGGTTGCAGG ACAAATGCTT CAGTCCACCG
951 AGAGCAGTAC CGTGTGGCCA AGAGGTGGAC TCAGAGCCTT CCTTGAGCTA
1001 AACTCGGCCA ACCAAGGCAC GCAGCATGTC CCCTCAGGTC TCCAGTCAGT
1051 CCAGGTTGAC CCTCAGTTCT GGACGTGTGT ATATAGCTGT ATTTAATACC
1101 TCAAGGTCAT TGTGGCTCTG GGGATGCCGG GGCAGGAGGA CGAGGGTGCG
1151 CTGTGGACAC AGCAGTCCGC GGAATTCCGT TCTGGGAAGC CAATGGTCGC
1201 CGGCACCCCT TGCTTCCTCC CTCTGTTGTC TGCCTGTGTG ACACACATCA
1251 ATGGCAATAA CTTCTTCCAA CTCCTCGCAG AAGTGGGAGA GGCCGGCAGC
1301 CTGCACCGAG AGGGACTTTC CTCTCTCTTG CTCCCCGCTT CGTTCTGTTT
1351 TGGCTGCAGA GAGTGGTTCA TCCATACTCT CATTCCCTCG CCTCCCCTTG
1401 TGGACGGGGG TCTTGCCTTT TCAATTCCTG TGTTTTGGTG TCTTCCCTTA
1451 TCTGCTACCC TGAATCACCT GTCCTGGTCT TGCTGTGTGA TGGGAACATG
1501 CTTGTAAACT GCGTAACAAA TCTACTTTGT GTATGTGTCT GTTTATGGGG
1551 GTGGTTTATT ATTTTTGCTG GTCCCTAGAC CACTTTGTAT GACCGTTTGC
1601 AGTCTGAGCA GGCCAGGGGC TGACAGCTAA TGTCAGGACC CTCAGCGGTG
1651 GAGCCTGCTG GGGGGACCCA GCTGCTCTTG GACAAGTGGC TGAGCTCCTA
1701 TCTGGCCTCC TCTTTTTTTT TTTTTCAAGT AATTTGTGTG TATTTCTAAC
1751 TGATGTATTG AAAAAATTCC TAGTATTTCA GTAAAAATGC CTGTTGTGAG
1801 ATGAACCTCC TGTAACTTCT ATCTGTTCTT TTTTGAGGCT CAGGGAGAAA
1851 CTAGCATTTT TTTTTTCCAA ACTACTTTTT GTCACTGTGA CAGTTGTAAA
1901 TAAAGTTTGA AAATGCTTTC CAAAAAAAAA AAAAAAAAA
B: aminoacid sequence (SEQ ID NO:26) length: 127 amino acid
1 MPGQEDEGAL WTQQSAEFRS GKPMVAGTPC FLPLLSACVT HINGNNFFQL
51 LAEVGEAGSL HREGLSSLLL PASFCFGCRE WFIHTLIPSP PLVDGGLAFS
101 IPVFWCLPLS ATLNHLSWSC CVMGTCL
C: Nucleotide and amino acid composite sequence (SEQ ID NO:27)
Clone number and protein name: PP2281
Start code: 1124 ATG stop coding: 1507 TAA
Protein molecular weight: 13698.16
1 C AGA CAT TCA CTC TGG CTG CTG GGA CAT CAG AAA ACA AAG TCT TCA 46
47 TCT CTC TCT CCA GTT TCA CCC ACC CCA CCC TTT GCT TTC ATT TCA GGT 94
95 GTG TTG GTC TAT ATG ACA GGG AGG AGA GTA AAG GAG AGC AGG AGC AAT 142
143 TGG CTG CCT GCA AAG CCA GCT GGA GGT GAA GTG CAG GAA AGG AAA GGT 190
191 CAC CCC ATT CTA CTC CAT GGC CTC TCT GCT CCC AGC TGT GGT AGG CTC 238
239 ACA TAG CCA GTG TGA TCG GTT TTT AAG AGG CAG TGC TTT TCA GCT TTT 286
287 CTC CCT GAT ATA TCC ATT TTG CTT CCC AGC ACT TTT TAG GAG TAG TGA 334
335 GAG CGC TTC CTG CCC TTG TTG GAA GCC CCA GGG TGG ACT CTC AGC ACG 382
383 AAG GTC TCT CCC TTA ACT GCT GCC CTT CCA AGA CTT GCT CCC GAG ATG 430
431 GAG TGG GCG TGG TCT TCC AGG CTG GCC CTT CCT TCT CCT CAC CGC CAC 478
479 CTT CCC TGC CCC AGC CCC AGC AGC CAT GGG TAC ATG GGT CCC CAG CTC 526
527 ACC TAT GGA TTC CCG CCA GTC TGC CCA GCT GCA GTA CTC ACG CCC CAT 574
575 GGG GGA TCT TGG TCT GTT TTT CTT GTG GGA GCC TAG TGG AGA GCA GAC 622
623 GTG GCT TTT TAT GTG TCT TGT TGG GGA GGT GAC TTG CAT GGT GGG GAC 670
671 AAG GCT GTC GTG GCA ACC TTG GGA TCG AGT TTG AGA CTA AAG GAT GTC 718
719 ATG AGA TCC CTG GCT TCT CCC CAT GTT GTT CCC GGA CAA GGG GAG AAG 766
767 GGA GGC ATG GCA AGG GAC CTC TGC TGT CCT TAC TCA ACA GTG GTC CTC 814
815 ATC CCT CCC CAC CTC CCA CTG CTT CCT CCA AGG GCA CCA GTT GTA TGA 862
863 GAA AGT TGG CCT TTG GAC TTA GGA TTT CTT ATT GTA GCT AAG AGC CAT 910
911 CTG AAG CAG CAG GTT GCA GGA CAA ATG CTT CAG TCC ACC GAG AGC AGT 958
959 ACC GTG TGG CCA AGA GGT GGA CTC AGA GCC TTC CTT GAG CTA AAC TCG 1006
1007 GCC AAC CAA GGC ACG CAG CAT GTC CCC TCA GGT CTC CAG TCA GTC CAG 1054
1055 GTT GAC CCT CAG TTC TGG ACG TGT GTA TAT AGC TGT ATT TAA TAC CTC 1102
1103 AAG GTC ATT GTG GCT CTG GGG ATG CCG GGG CAG GAG GAC GAG GGT GCG 1150
1 Met Pro Gly Gln Glu Asp Glu Gly Ala 9
1151 CTG TGG ACA CAG CAG TCC GCG GAA TTC CGT TCT GGG AAG CCA ATG GTC 1198
10 Leu Trp Thr Gln Gln Ser Ala Glu Phe Arg Ser Gly Lys Pro Met Val 25
1199 GCC GGC ACC CCT TGC TTC CTC CCT CTG TTG TCT GCC TGT GTG ACA CAC 1246
26 Ala Gly Thr Pro Cys Phe Leu Pro Leu Leu Ser Ala Cys Val Thr His 41
1247 ATC AAT GGC AAT AAC TTC TTC CAA CTC CTC GCA GAA GTG GGA GAG GCC 1294
42 Ile Asn Gly Asn Asn Phe Phe Gln Leu Leu Ala Glu Val Gly Glu Ala 57
1295 GGC AGC CTG CAC CGA GAG GGA CTT TCC TCT CTC TTG CTC CCC GCT TCG 1342
58 Gly Ser Leu His Arg Glu Gly Leu Ser Ser Leu Leu Leu Pro Ala Ser 73
1343 TTC TGT TTT GGC TGC AGA GAG TGG TTC ATC CAT ACT CTC ATT CCC TCG 1390
74 Phe Cys Phe Gly Cys Arg Glu Trp Phe Ile His Thr Leu Ile Pro Ser 89
1391 CCT CCC CTT GTG GAC GGG GGT CTT GCC TTT TCA ATT CCT GTG TTT TGG 1438
90 Pro Pro Leu Val Asp Gly Gly Leu Ala Phe Ser Ile Pro Val Phe Trp 105
1439 TGT CTT CCC TTA TCT GCT ACC CTG AAT CAC CTG TCC TGG TCT TGC TGT 1486
106 Cys Leu Pro Leu Ser Ala Thr Leu Asn His Leu Ser Trp Ser Cys Cys 121
1487 GTG ATG GGA ACA TGC TTG TAA ACT GCG TAA CAA ATC TAC TTT GTG TAT 1534
122 Val Met Gly Thr Cys Leu *** 128
1535 GTG TCT GTT TAT GGG GGT GGT TTA TTA TTT TTG CTG GTC CCT AGA CCA 1582
1583 CTT TGT ATG ACC GTT TGC AGT CTG AGC AGG CCA GGG GCT GAC AGC TAA 1630
1631 TGT CAG GAC CCT CAG CGG TGG AGC CTG CTG GGG GGA CCC AGC TGC TCT 1678
1679 TGG ACA AGT GGC TGA GCT CCT ATC TGG CCT CCT CTT TTT TTT TTT TTC 1726
1727 AAG TTT TTT GTG TGT ATT TCT AAC TGA TGT ATT GAA AAA ATT CCT AGT 1774
1775 ATT TCA GTA AAA ATG CCT GTT GTG AGA TGA ACC TCC TGT AAC TTC TAT 1822
1823 CTG TTC TTT TTT GAG GCT CAG GGA GAA ACT AGC ATT TTT TTT TTC CAA 1870
1871 ACT ACT TTT TGT CAC TGT GAC AGT TGT AAA TAA AGT TTG AAA ATG CTT 1918
1919 TCC AAA AAA AAA AAA AAA AAA 1939
10.PP2447
A: nucleotide sequence (SEQ ID NO:28) length: 1678bp
1 CCATGTTGGC CAGGCTGGTC TGAGAGAGAA CTCCTGACCT CGTGATCCGC
51 CTGCCTCAGC CTCCCAAAGT GCTGGGATGA CAGGCGTGCG CCACTGCACC
101 CGGCCCGTGA AGGTCTTTTC TGAAGGGAGC TACCTTTGAG AAGTGCCTGT
151 TGGGACAGAA CTCCCTGCTC TGGTCTGACC CTCTGCACGC TAGGGTGGTC
201 CTGCACCTGT CGGGGAGGGG GCCAAGGACA CAGCACTGGG CTGGGGCAGG
251 GCTCTGCGCT GGTGCTGCCT GCCATCCCAG TTCTGCCATT CCCCCAGCGC
301 ACCCCCGCAC CCGCCCCTCC AGCTGATGCC TGCTCCCTCT CTCTGCAGAA
351 CGGGCTCATG TCGGGGCTGA TGCAGATGCT GCTGCTGAAG GTGTCTGCAC
401 ACATCACCGA GCAGCTGGGC ATGGCCCCAG GTGGCGAGTT CAGGGAGGCC
451 TTCAAGGAGG TGGGCACAGG GTGAGGTAGG GGGTGGCCAC AGGGATGTGG
501 CTCACAGGGG TGGTCCGGGG TTCCCAGTGC TCCCCAACAC CCAGCCTCTG
551 CTCCAGGCCA GCAAGGTGCC TTTCTGCAAG TTCCACCTGG GTGACCGACC
601 CATCCCCGTC ACCTTCAAGA GGGCCATCGC AGCGCTCTCC TTCTGGCAGA
651 AGGTCAGGCT GGCTTGGGGC CTGTGCTTCC TGTCAGACCC CATCAGGTAG
701 GGCTGCCCCC GGGACCCTGG CCGGCCTGCA GGGTGGTCTG TGGGAGGCTC
751 CAGGCCCTCC TGTGCAGGTC CAAGCGCAGC CAATCCTCAC TCAAGGCCTT
801 CCCTGCCCTT TCCTTCCGCC ACAAATCCCA AACAAACGTG CTGTGGTCCC
851 TGCCCGGTGT CCACAGTGCC AGCCCCACCC CCCCAGCCCG TTGCCCATCC
901 CTGCGGGGCT GCAGCCATCC CTCTCCACAG CAAGGATGAC GTGGAACGCT
951 GCAAGCAGAA GGACCTACTG GAGCAGATGA TGGCCGAGAT GATTGGCGAG
1001 TTCCCAGACC TGCACCGCAC CATCGTCTCG GAGCGCGACG TCTACCTAAC
1051 CTACATGCTG CGCCAGGCCG CGCGGCGCCT CGAGCTGCCT CGGGCCTCTG
1101 ACGGTGACGG CCGCCCGCAG GCGTGGGACC CCCTGTGAGG CTGAGGCCCG
1151 AGCAGGTACT GACCCCTTGT CCTTCCCCAC AGCCGAGCCC AGGAAGTGCG
1201 TCCCCTCCGT GGTCGTGGGC GTCGTGGGCA TGGGCCACGT GCCTGGCATC
1251 GAGAAGAACT GGAGCACCGA CCTCAACATC CAGGAGATCA TGACCGTGCC
1301 CCCGCCGTCC GTCTCCGGCA GAGTGTCTCG GTTGGCCGTG AAGGCCGCCT
1351 TCTTCGGCCT GCTGGGCTAC AGCCTGTACT GGATGGGCCG CCGCACCGCG
1401 AGCCTGGTCC TGTCGCTGCC CGCCGCGCAG TACTGCCTGC AGAGGGTGAC
1451 CGAGGCCCGG CACAAGTAGG AGACTGCTCC CCGCCCGCTC GGGCCCCTGA
1501 GGAGCCAGTG CCCCCGCGGC ACTTCTGGCT GCCAGGTGCA TCCTAGCCCG
1551 CCCGAGGCCC CTGCCACCCC CCATGGGGGT CTGGGCCCGG CCTCGCCTGC
1601 CCTCCTGGGC CAGTCACCCC TCCCCCAGCC CACCCAAATA AAGGATTATT
1651 TAACTGTCTG AAAAAAAAAA AAAAAAAA
B: aminoacid sequence (SEQ ID NO:29) length: 161 amino acid
1 MWLTGVVRGS QCSPTPSLCS RPARCLSASS TWVTDPSPSP SRGPSQRSPS
51 GRRSGWLGAC ASCQTPSGRA APGTLAGLQG GLWEAPGPPV QVQAQPILTQ
101 GLPCPFLPPQ IPNKRAVVPA RCPQCQPHPP SPLPIPAGLQ PSLSTARMTW
151 NAASRRTYWS R
C: Nucleotide and amino acid composite sequence (SEQ ID NO:30)
Clone number and protein name: PP2447
Start code: 495 ATG stop coding: 980 TGA
Protein molecular weight: 16922.45
1 CC ATG TTG GCC AGG CTG GTC TGA GAG AGA ACT CCT GAC CTC GTG ATC 47
48 CGC CTG CCT CAG CCT CCC AAA GTG CTG GGA TGA CAG GCG TGC GCC ACT 95
96 GCA CCC GGC CCG TGA AGG TCT TTT CTG AAG GGA GCT ACC TTT GAG AAG 143
144 TGC CTG TTG GGA CAG AAC TCC CTG CTC TGG TCT GAC CCT CTG CAC GCT 191
192 AGG GTG GTC CTG CAC CTG TCG GGG AGG GGG CCA AGG ACA CAG CAC TGG 239
240 GCT GGG GCA GGG CTC TGC GCT GGT GCT GCC TGC CAT CCC AGT TCT GCC 287
288 ATT CCC CCA GCG CAC CCC CGC ACC CGC CCC TCC AGC TGA TGC CTG CTC 335
336 CCT CTC TCT GCA GAA CGG GCT CAT GTC GGG GCT GAT GCA GAT GCT GCT 383
384 GCT GAA GGT GTC TGC ACA CAT CAC CGA GCA GCT GGG CAT GGC CCC AGG 431
432 TGG CGA GTT CAG GGA GGC CTT CAA GGA GGT GGG CAC AGG GTG AGG TAG 479
480 GGG GTG GCC ACA GGG ATG TGG CTC ACA GGG GTG GTC CGG GGT TCC CAG 527
1 Met Trp Leu Thr Gly Val Val Arg Gly Ser Gln 11
528 TGC TCC CCA ACA CCC AGC CTC TGC TCC AGG CCA GCA AGG TGC CTT TCT 575
12 Cys Ser Pro Thr Pro Ser Leu Cys Ser Arg Pro Ala Arg Cys Leu Ser 27
576 GCA AGT TCC ACC TGG GTG ACC GAC CCA TCC CCG TCA CCT TCA AGA GGG 623
28 Ala Ser Ser Thr Trp Val Thr Asp Pro Ser Pro Ser Pro Ser Arg Gly 43
624 CCA TCG CAG CGC TCT CCT TCT GGC AGA AGG TCA GGC TGG CTT GGG GCC 671
44 Pro Ser Gln Arg Ser Pro Ser Gly Arg Arg Ser Gly Trp Leu Gly Ala 59
672 TGT GCT TCC TGT CAG ACC CCA TCA GGT AGG GCT GCC CCC GGG ACC CTG 719
60 Cys Ala Ser Cys Gln Thr Pro Ser Gly Arg Ala Ala Pro Gly Thr Leu 75
720 GCC GGC CTG CAG GGT GGT CTG TGG GAG GCT CCA GGC CCT CCT GTG CAG 767
76 Ala Gly Leu Gln Gly Gly Leu Trp Glu Ala Pro Gly Pro Pro Val Gln 91
768 GTC CAA GCG CAG CCA ATC CTC ACT CAA GGC CTT CCC TGC CCT TTC CTT 815
92 Val Gln Ala Gln Pro Ile Leu Thr Gln Gly Leu Pro Cys Pro Phe Leu 107
816 CCG CCA CAA ATC CCA AAC AAA CGT GCT GTG GTC CCT GCC CGG TGT CCA 863
108 Pro Pro Gln Ile Pro Asn Lys Arg Ala Val Val Pro Ala Arg Cys Pro 123
864 CAG TGC CAG CCC CAC CCC CCC AGC CCG TTG CCC ATC CCT GCG GGG CTG 911
124 Gln Cys Gln Pro His Pro Pro Ser Pro Leu Pro Ile Pro Ala Gly Leu 139
912 CAG CCA TCC CTC TCC ACA GCA AGG ATG ACG TGG AAC GCT GCA AGC AGA 959
140 Gln Pro Ser Leu Ser Thr Ala Arg Met Thr Trp Asn Ala Ala Ser Arg 155
960 AGG ACC TAC TGG AGC AGA TGA TGG CCG AGA TGA TTG GCG AGT TCC CAG 1007
156 Arg Thr Tyr Trp Ser Arg *** 162
1008 ACC TGC ACC GCA CCA TCG TCT CGG AGC GCG ACG TCT ACC TAA CCT ACA 1055
1056 TGC TGC GCC AGG CCG CGC GGC GCC TCG AGC TGC CTC GGG CCT CTG ACG 1103
1104 GTG ACG GCC GCC CGC AGG CGT GGG ACC CCC TGT GAG GCT GAG GCC CGA 1151
1152 GCA GGT ACT GAC CCC TTG TCC TTC CCC ACA GCC GAG CCC AGG AAG TGC 1199
1200 GTC CCC TCC GTG GTC GTG GGC GTC GTG GGC ATG GGC CAC GTG CCT GGC 1247
1248 ATC GAG AAG AAC TGG AGC ACC GAC CTC AAC ATC CAG GAG ATC ATG ACC 1295
1296 GTG CCC CCG CCG TCC GTC TCC GGC AGA GTG TCT CGG TTG GCC GTG AAG 1343
1344 GCC GCC TTC TTC GGC CTG CTG GGC TAC AGC CTG TAC TGG ATG GGC CGC 1391
1392 CGC ACC GCG AGC CTG GTC CTG TCG CTG CCC GCC GCG CAG TAC TGC CTG 1439
1440 CAG AGG GTG ACC GAG GCC CGG CAC AAG TAG GAG ACT GCT CCC CGC CCG 1487
1488 CTC GGG CCC CTG AGG AGC CAG TGC CCC CGC GGC ACT TCT GGG TGC CAG 1535
1536 GTG CAT CCT AGC CCG CCC GAG GCC CCT GCC ACC CCC CAT GGG GGT CTG 1583
1584 GGC CCG GCC TCG CCT GCC CTC CTG GGC CAG TCA CCC CTC CCC CAG CCC 1631
1632 ACC CAA ATA AAG GAT TAT TTA ACT GTC TGA AAA AAA AAA AAA AAA AA 1678
11.PP3111
A: nucleotide sequence (SEQ ID NO:31) length: 1034bp
1 AGGACGTGGA GCGCTGCCTC CGGGACACGG GTGTGCAGGG CGTCATGAGC
51 GCAGAGGGCA ACCTGCACAA CCCCGCCCTG TTCGAGGGCC GGAGCCCTGC
101 CGTGTGGGAG CTGGCCGAGG AGTATCTGGA CATCGTGCGG GAGCACCCCT
151 GCCCCCTGTC CTACGTCCGG GCCCACCTCT TCAAGCTGTG GCACCACACG
201 CTGCAGGTGC ACCAGGAGCT GCGAGAGTAG CTGGCCAAGG TGAAGACCCT
251 GGAGGGCATC GCTGCTGTGA GCCAGGAGCT GAAGCTGCGG TGTCAGGAGG
301 AGATATCCAG GCAGGAGGGA GCGAAGCCCA CCGGCGACTT GCCCTTCCAC
351 TGGATCTGCC AGCCCTACAT CCGGCCGGGG CCCAGGGAGG GGAGCAAGGA
401 GAAGGCAGGT GCGCGCAGCA AGCGGGCCCT GGAGGAAGAG GAGGGTGGCA
451 CGGAGGTCCT GTCCAAGAAC AAGCAAAAGA AGCAGCTGAG GAACCCCCAC
501 AAGACCTTCG ACCCCTCTCT GAAGCCAAAA TATGCAAAGT GTGACCAGTG
551 TGGAAACCCA AAGGGCAACA GATGTGTGTT CAGCCTGTGC CGCGGCTGCT
601 GCAAGAAGCG AGCCTCCAAA GAGACTGCAG ACTGCCCAGG TCACGGATTG
651 CTTTTTAAAA CCAAATTGGA GAAGTCTCTG GCCTGGAAAG AGGCCCAGCC
701 TGAGCTGCAG GAGCCTCAGC CAGCAGCACC TGGAACACCA GGTGGCTTCT
751 CCGAAGTCAT GGGCAGTGCC CTGGCCTGAA GGCCCACAAC CCCCACCCCC
801 AGGACTGCTG CTGGAGCCTG GACACGTCCT ACTTAAGAAA ATGCCTTTTA
851 CTCAGGGAAT CTCCTGCTAC TTAATGTGGA AAGACACGCC CATGTCCCCC
901 TTCGGCCCAC TCTGGGGGCC TGGAAATGCT GCAGTGGGGA GCAGGCCCCA
951 GGCTGGACCT GCCCTGTCCT CAGCACGCGT GTGCAAAAGT GAACAATAAA
1001 TCATTTCAAA GATGCGAAAA AAAAAAAAAA AAAA
B: aminoacid sequence (SEQ ID NO:32) length: 155 amino acid
1 MQSVTSVETQ RATDVCSACA AAAARSEPPK RLQTAQVTDC FLKPNWRSLW
51 PGKRPSLSCR SLSQQHLEHQ VASPKSWAVP WPEGPQPPPP GLLLEPGHVL
101 LKKMPFTQGI SCYLMWKDTP MSPFGPLWGP GNAAVGSRPQ AGPALSSARV
151 CKSEQ
C: Nucleotide and amino acid composite sequence (SEQ ID NO:33)
Clone number and protein name: PP3111
Start code: 532 ATG stop coding: 999 TAA
Protein molecular weight: 16780.42
1 AGG ACG TGG AGC GCT GCC TCC GGG ACA CGG GTG TGC AGG GCG TCA TGA 48
49 GCG CAG AGG GCA ACC TGC ACA ACC CCG CCC TGT TCG AGG GCC GGA GCC 96
97 CTG CCG TGT GGG AGC TGG CCG AGG AGT ATC TGG ACA TCG TGC GGG AGC 144
145 ACC CCT GCC CCC TGT CCT ACG TCC GGG CCC ACC TCT TCA AGC TGT GGC 192
193 ACC ACA CGC TGC AGG TGC ACC AGG AGC TGC GAG AGT AGC TGG CCA AGG 240
241 TGA AGA CCC TGG AGG GCA TCG CTG CTG TGA GCC AGG AGC TGA AGC TGC 288
289 GGT GTC AGG AGG AGA TAT CCA GGC AGG AGG GAG CGA AGC CCA CCG GCG 336
337 ACT TGC CCT TCC ACT GGA TCT GCC AGC CCT ACA TCC GGC CGG GGC CCA 384
385 GGG AGG GGA GCA AGG AGA AGG CAG GTG CGC GCA GCA AGC GGG CCC TGG 432
433 AGG AAG AGG AGG GTG GCA CGG AGG TCC TGT CCA AGA ACA AGC AAA AGA 480
481 AGC AGC TGA GGA ACC CCC ACA AGA CCT TCG ACC CCT CTC TGA AGC CAA 528
529 AAT ATG CAA AGT GTG ACC AGT GTG GAA ACC CAA AGG GCA ACA GAT GTG 576
1 Met Gln Ser Val Thr Ser Val Glu Thr Gln Arg Ala Thr Asp Val 15
577 TGT TCA GCC TGT GCC GCG GCT GCT GCA AGA AGC GAG CCT CCA AAG AGA 624
16 Cys Ser Ala Cys Ala Ala Ala Ala Ala Arg Ser Glu Pro Pro Lys Arg 31
625 CTG CAG ACT GCC CAG GTC ACG GAT TGC TTT TTA AAA CCA AAT TGG AGA 672
32 Leu Gln Thr Ala Gln Val Thr Asp Cys Phe Leu Lys Pro Asn Trp Arg 47
673 AGT CTC TGG CCT GGA AAG AGG CCC AGC CTG AGC TGC AGG AGC CTC AGC 720
48 Ser Leu Trp Pro Gly Lys Arg Pro Ser Leu Ser Cys Arg Ser Leu Ser 63
721 CAG CAG CAC CTG GAA CAC CAG GTG GCT TCT CCG AAG TCA TGG GCA GTG 768
64 Gln Gln His Leu Glu His Gln Val Ala Ser Pro Lys Ser Trp Ala Val 79
769 CCC TGG CCT GAA GGC CCA CAA CCC CCA CCC CCA GGA CTG CTG CTG GAG 816
80 Pro Trp Pro Glu Gly Pro Gln Pro Pro Pro Pro Gly Leu Leu Leu Glu 95
817 CCT GGA CAC GTC CTA CTT AAG AAA ATG CCT TTT ACT CAG GGA ATC TCC 864
96 Pro Gly His Val Leu Leu Lys Lys Met Pro Phe Thr Gln Gly Ile Ser 111
865 TGC TAC TTA ATG TGG AAA GAC ACG CCC ATG TCC CCC TTC GGC CCA CTC 912
112 Cys Tyr Leu Met Trp Lys Asp Thr Pro Met Ser Pro Phe Gly Pro Leu 127
913 TGG GGG CCT GGA AAT GCT GCA GTG GGG AGC AGG CCC CAG GCT GGA CCT 960
128 Trp Gly Pro Gly Asn Ala Ala Val Gly Ser Arg Pro Gln Ala Gly Pro 143
961 GCC CTG TCC TCA GCA CGC GTG TGC AAA AGT GAA CAA TAA ATC ATT TCA 1008
144 Ala Leu Ser Ser Ala Arg Val Cys Lys Ser Glu Gln *** 156
1009 AAG ATG CGA AAA AAA AAA AAA AAA AA 1034

Claims (5)

1. isolating polynucleotide is characterized in that, it comprises a nucleotide sequence, and this nucleotide sequence is selected from down group:
(a) coding has the proteic polynucleotide of people of cancer suppressing function, and described albumen has 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、SEQ ID NO：23、SEQ ID NO：26、SEQ ID NO：29、SEQ IDNO：32；
(b) with polynucleotide (a) complementary polynucleotide.
2. polynucleotide as claimed in claim 1, 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 IDNO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:26, SEQ IDNO:29, SEQ ID NO:32.
3. polynucleotide as claimed in claim 1 is characterized in that, the sequence of these polynucleotide is selected from down group:
Coding region sequence or the full length sequence of SEQ ID NO: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, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQID NO:33.
4. a carrier is characterized in that, it contains the described polynucleotide of claim 1.
5. a genetically engineered host cell is characterized in that, it is a kind of host cell that is selected from down group:
(a) host cell that transforms or transduce with the described carrier of claim 4;
(b) host cell that transforms or transduce with the described polynucleotide of claim 1.
CNB001119494A 2000-03-09 2000-03-09 Human protein able to suppress growth of cancer cells and its coding sequence Expired - Fee Related CN1169955C (en)

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