CN1286698A - Human angiotonin II/vasopressin receptor like gene - Google Patents

Human angiotonin II/vasopressin receptor like gene Download PDF

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CN1286698A
CN1286698A CN98813892.1A CN98813892A CN1286698A CN 1286698 A CN1286698 A CN 1286698A CN 98813892 A CN98813892 A CN 98813892A CN 1286698 A CN1286698 A CN 1286698A
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polypeptide
cbdakd01
sequence
polynucleotide
nucleotide sequence
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张庆华
沈宇
茅矛
顾柏炜
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Shanghai Second Medical University
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Shanghai Second Medical University
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    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies

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  • General Health & Medical Sciences (AREA)
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Abstract

CBDAKD01 polpeptides and polynucleiotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing CBDAKD01 polypeptides and polynucleotides in the design of protocols for the treatment of hypertension, heart disease, cancer, and kidney disease, among others, and diagnostic assays for such conditions.

Description

Human angiotensin II/II/vasopressin receptor like gene
Invention field
The present invention relates to new identified polynucleotides, by the purposes and the production thereof of its encoded polypeptides and these polynucleotide and polypeptide.More specifically, polynucleotide of the present invention and polypeptide and Angiotensin and/(hereinafter referred to as CBDAKD01) is relevant in II/vasopressin receptor family.The invention still further relates to the effect that suppresses or activate these polynucleotide and polypeptide.
Background of invention
Rat serum angiotensin/II/vasopressin receptor (AII/AVP) is a dual receptor, and it can be in conjunction with angiotensin and beta-hypophamine.This shows that Angiotensin and/or II/vasopressin receptor family are as the existing definite precedent that confirms of treatment target.Obviously, other member of working aspect prevention, alleviation and debugging functions obstacle or the disease, described dysfunction or disease include but not limited to hypertension, heart trouble, cancer and ephrosis in needs evaluation and sign Angiotensin and/or the II/vasopressin receptor family.
Summary of the invention
One aspect of the present invention relates to CBDAKD01 polypeptide and recombined material and their production method.On the other hand, the present invention relates to utilize the method for CBDAKD01 polypeptide and polynucleotide, comprise diseases such as treatment hypertension, heart trouble, cancer and ephrosis.Again on the one hand, the present invention relates to utilize the method for material evaluation agonist provided by the invention and antagonist and utilize the symptom that compounds identified is treated and CBDAKD01 is uneven relevant.At last, the present invention relates to check the diagnostic test of the disease relevant with CBDAKD01 activity or horizontal abnormality.The detailed Description Of The Invention definition
For ease of understanding term commonly used in this specification sheets, provide to give a definition.
" CBDAKD01 " generally is meant polypeptide or its allelic variant with aminoacid sequence shown in the sequence 2.
" CBDAKD01 activity or CBDAKD polypeptide active " or " CBDAKD01 or CBDAKD01 polypeptide biological activity " is meant metabolism or the physiological function of CBDAKD01, comprises the active of similar activity or raising or has the activity of hanging down undesirable side effects.Described activity also comprises antigen and the immunogen activity of CBDAKD01.
" CBDAKD01 gene " is meant polynucleotide or its allelic variant and/or its complement with nucleotide sequence shown in the sequence 1.
" antibody " comprises mono-clonal and polyclonal antibody, chimeric, strand and humanized antibody and Fab fragment in this article, comprises the product in Fab or other immunoglobulin expression libraries.
" isolating " expression " by artificial " changes its native state.If " isolating " composition or material exist at occurring in nature, then it changes or takes out or taken out and change from its primal environment.For example, when using this term in this article, polynucleotide or the polypeptide that exists in moving object is not " isolating ", but with native state under the material that coexists the as much Nucleotide or the polypeptide that separate be exactly " isolating ".
" polynucleotide " refer generally to poly-ribonucleotide or poly-deoxyribonucleotide, can be the RNA or the DNA of unmodified or modification." polynucleotide " include but not limited to strand and double-stranded DNA, strand and double stranded region blended DNA, strand and double-stranded RNA, strand and double stranded region blended RNA, the hybrid molecule that contains DNA and RNA can be strand or more common mixture for two strands or strand and double stranded region.In addition, " polynucleotide " also refer to contain three sequences of RNA or DNA or RNA and DNA.Term " polynucleotide " also comprises DNA or the RNA that contains one or more modified bases, and is DNA or RNA that stability or other reasons have been modified skeleton." modification " base comprises for example rare base such as tritylation base and inosine.Can carry out multiple modification to DNA and RNA, therefore, " polynucleotide " comprise chemistry, enzymatic or the metabolism modified forms of the polynucleotide that occurring in nature is common, and the DNA and the RNA chemical species of virus and cells characteristic." polynucleotide " also comprise short, that be commonly called oligonucleotide relatively polynucleotide.
" polypeptide " refers to that the peptide bond that contains by peptide bond or modification is interconnected two or more amino acid whose any peptides of peptide isostere (peptideisosteres) or albumen." polypeptide " both comprised the oligopeptides or the oligomer of short chain (being commonly referred to peptide), also comprised long-chain (being commonly referred to albumen).Polypeptide may contain the amino acid outside the amino acid of 20 kinds of genes encodings." polypeptide " comprises the aminoacid sequence of modifying by translation natural method such as post-treatment or chemical modification technology known in the art.It is detailed that these are modified in basic textbook and monograph and a large amount of research paper narration.Modification can occur in any position of polypeptide, comprises peptide backbone, amino acid side chain and amino or C-terminal.Can exist with identical or different degree in the similar modification of the different loci of given polypeptide.And given polypeptide can contain polytype modification.Polypeptide can be owing to ubiquitination branch, also can be branch or unbranched ring-type.Ring-type, branch and branch's ring type polypeptide may be because natural process after the translation or synthetic method generations.Modification comprises acetylize, acidylate, the ADP-ribosylation, amidation, the covalency of flavine stops and closes, the covalency of protoheme stops and closes, the covalency of Nucleotide or nucleotide derivative stops and closes, the covalency of fat or fat derivative stops and closes, the covalency of phosphatidylinositols stops and closes, crosslinked, cyclisation, disulfide linkage forms, demethylation, the formation of covalent cross-linking, Gelucystine forms, Pyrrolidonecarboxylic acid forms, formylation, the γ carboxylation, glycosylation, the GPI deadman forms, hydroxylation, iodate, methylate, the Semen Myristicae acidylate, oxidation, proteolysis processing, phosphorylation, isoamyl two acidylates, racemization, selenium acidylate (selenoylation), sulfation, the amino acid of transfer RNA mediation adds protein, (can be as arginylization and ubiquitination referring to for example " protein: structure and molecular characterization " the 2nd edition, T.E.Creighton, W.H.Freeman and Company, New York, 1993 and Wold, F., " posttranslational protein is modified: prospect and prospect ", the 1-12 page or leaf, " proteic translation back covalent modification " B.C.Johnson, Ed., Academic Press, NewYork, 1983; Seifter et al., " analysis of protein modified and non-albumen cofactor ", MethEnzymol (1990) 82:626-646 and Rattan et al., " protein synthesis: posttranslational modification and aging ", Ann NY Acad Sci (1992) 663:48-62).
" variant " refers to be different from reference to polynucleotide or polypeptide but kept the polynucleotide or the polypeptide of fundamental characteristics.The nucleotide sequence of general polynucleotide variant is with different with reference to polynucleotide.The variation of variant nucleotide sequence may change or not change the amino acid sequence of polypeptide by the reference polynucleotide encoding.As described below, Nucleotide changes may cause amino acid whose displacement, interpolation, disappearance, fusion and brachymemma in the canonical sequence encoded polypeptides.The aminoacid sequence of general polypeptide variants is with different with reference to polypeptide.Usually, difference is limited, and therefore the sequence with reference to polypeptide and variant is extremely proximate on the whole, is identical in many zones.Variant and one or more displacements, the interpolation of any array configuration, the difference of disappearance can be arranged on aminoacid sequence with reference to polypeptide.Displacement or the amino-acid residue that inserts can yes or no genetic code amino acids coding.Polynucleotide or variant polypeptides can be naturally occurring, as allelic variant, or the non-natural existence.Polynucleotide that non-natural exists and polypeptide variants can be by induced-mutation technique or directly synthetic preparations.
" identity " is known in the art is the two or more peptide sequences determined by comparative sequences or the relation between the polynucleotide sequence.In the art, " identity " also represents the polypeptide determined by the coupling between these sequence chains or the sequence degree of correlation between the polynucleotide sequence." identity " and " similarity " can be calculated easily by currently known methods, includes but not limited to method described in the following document: " calculating molecular biology ", Lesk, A.M., ed.OxfordUniversity Press, New York, 1988; " calculation biology: brief introduction and genome project ", Smith D.W., ed., Academic Press, New York, 1993; " Computer Analysis of sequence data " part 1, Griffin A.M., and Griffin H.G., eds., Humana Press, NewJersey, 1994; " sequential analysis in the molecular biology ", von Heiinje, G., Academic Press, 1987; " sequence analysis primer ", Gribskov, M.and Devereux, J., eds., M StocktonPress, NeW York, 1991; Carillo, H., and Lipman, D., SIAM J.Applied Math., 48:1073 (1988).The preferred method of determining identity is designed to make between the sequence of test coupling maximum.And, determine that the method for identity and similarity has been compiled to the computer program that can openly obtain.Determine two between the sequence identity and the preferred computer program technic of similarity include but not limited to GCG routine package (Devereux, J., et al., Nucleic Acids Research (1984) 12 (1): 387), BLASTP, BLASTN, FASTA (Atschul, S.F.et al., J Molec Biol (1990) 215:403).BLAST X program can obtain from NCBI or other source (the BLAST handbook, Altschul, S., et al., NCBI NLM NIH Bethesda, MD 20894; Altschul, S., etal., J.Mol.Biol.215:403-410 (1990).Well-known Smith Waterman algorithm also can be used for determining identity.
The preferred parameter of peptide sequence comparison comprises: 1) algorithm: Needleman and Wunsch, J.Mol Biol.48:443-453 (1970) comparator matrix (Comparison matrix): BLOSSUM62, from Hentikoff andHentikoff, the compensation of Proc.Natl.Acad.Sci.USA.89: 10915-10919 (1992) breach: the compensation of 12 notch length: 4 programs can using these parameters are Genetics Computers Group, the gap program that Madison WI provides.Above-mentioned parameter is a peptide default parameter (terminal breach uncompensation) relatively.
The preferred parameter of polynucleotide comparison comprises: 1) algorithm: Needleman and Wunsch, J.Mol Biol.48:443-453 (1970) comparator matrix (Comparison matrix): coupling=+ 10, mispairing=0 breach compensation: 50 notch length compensation: 3 can to use a program of these parameters are Genetics Computers Group, the gap program that Madison WI provides.Above-mentioned parameter is a nucleic acid default parameter relatively.
The preferred polynucleotide embodiment comprises that also the identity that contains with the polynucleotide canonical sequence of sequence 1 is at least 50,60,70,80,85,90,95,97 or 100% separation polynucleotide, wherein said sequence can be identical with the canonical sequence of sequence 1, perhaps compare the Nucleotide that comprises some amount changes with canonical sequence, wherein said change is selected from least one nucleotide deletion, displacement (comprising conversion and transversion) or insertion, wherein said change can occur in reference to any position between 5 of polynucleotide sequence ' or 3 ' terminal or two ends, be dispersed in respectively between the Nucleotide that is distributed in canonical sequence, perhaps be dispersed in and be distributed in the canonical sequence with one or more contigs.The quantity that Nucleotide changes is following to be determined: the total nucleotide number in the sequence 1 multiply by corresponding identity percentage ratio, then its result is reduced from the total nucleotide number of sequence 1, perhaps:
n n≤X n-(X n·y)
N wherein nFor Nucleotide changes quantity, x nBe the total nucleotide number of sequence 1, the y value was 0.50 at 50% o'clock, was 0.60 at 60% o'clock, was 0.70 at 70% o'clock, at 80% o'clock was 0.80, was 0.85 at 85% o'clock, was 0.90 at 90% o'clock, was 0.95 at 95% o'clock, if at 97% o'clock was 0.97, was 1.00 at 100% o'clock, wherein x nWith the result of y be not integer, then it is rounded up and gets nearest integer, again from x nMiddle subduction.The change meeting of the polynucleotide sequence of encoding sequence 2 polypeptide produces nonsense, missense or phase shift mutation in its encoding sequence, thereby the polypeptide of the polynucleotide encoding after changing changes.
The preferred polypeptide embodiment comprises that also the identity that contains with the polypeptide canonical sequence of sequence 2 is at least 50,60,70,80,85,90,95,97 or 100% isolated polypeptide, wherein said sequence can be identical with the canonical sequence of sequence 2, perhaps compare the amino acid change that comprises some amount with canonical sequence, wherein said change can be selected from least one aminoacid deletion, displacement (comprising conservative and non-conservative substitution) or insertion, wherein said change can occur in reference to any position between the amino of peptide sequence or C-terminal or two ends, be dispersed in respectively between the amino acid that is distributed in canonical sequence, perhaps be dispersed in and be distributed in the canonical sequence with one or more contigs.The quantity of amino acid change is following to be determined: the total amino acid number in the sequence 2 multiply by corresponding identity percentage ratio, then its result is reduced from the total amino acid number of sequence 2, perhaps:
n a≤x a-(x a·y)
N wherein aBe amino acid change quantity, x aBe the total amino acid number of sequence 2, the y value was 0.50 at 50% o'clock, was 0.60 at 60% o'clock, was 0.70 at 70% o'clock, at 80% o'clock was 0.80, was 0.85 at 85% o'clock, was 0.90 at 90% o'clock, was 0.95 at 95% o'clock, if at 97% o'clock was 0.97, was 1.00 at 100% o'clock, wherein x aWith the result of y be not integer, then it is rounded up and gets nearest integer, again from x aMiddle subduction.Polypeptide of the present invention
On the one hand, the present invention relates to CBDAKD01 polypeptide (or CBDAKD01 albumen).The CBDAKD01 polypeptide comprises the polypeptide of sequence 2, and the polypeptide that contains the aminoacid sequence of sequence 2, with contain the polypeptide that identity with sequence 2 total lengths is at least 80% aminoacid sequence, preferably the identity with sequence 2 is at least 90%, more preferably identity is at least 95%.The sequence that identity is at least 97-99% is highly preferred.The CBDAKD01 polypeptide comprises that also its aminoacid sequence and the identity with polypeptide total length of sequence 2 aminoacid sequences are at least 80% polypeptide, and preferably the identity with sequence 2 is at least 90%, and more preferably identity is at least 95%.The sequence that identity is at least 97-99% is highly preferred.Preferred CBDAKD01 polypeptide has at least a CBDAKD01 biological activity.
The CBDAKD01 polypeptide can be " maturation " proteic form, also can be the part than large protein such as fusion rotein.It is comparatively favourable to generally include additional aminoacid sequence, and additional aminoacid sequence comprises secretion or leader sequence, and former sequence helps the sequence such as the polyhistidyl residue of purifying, or helps stable appended sequence in the recombinant production process.
The present invention also comprises the CBDAKD01 polypeptide fragment.Fragment is that the part of its aminoacid sequence and CBDAKD01 amino acid sequence of polypeptide is identical, and does not have whole aminoacid sequences of CBDAKD01.The same with the CBDAKD01 polypeptide, fragment can be independently, or is included in the bigger polypeptide as a part or a zone, preferably as an independent successive zone.The representative example of polypeptide fragment of the present invention comprises for example about fragment from amino acid/11-20,21-40,41-60,61-80,81-100 and 101 to CBDAKD01 polypeptide ends." approximately " is included in and appoints the more concrete described scope in one or both ends big or little several, 5,4,3,2 or 1 amino acid.
Preferred fragment comprises the polypeptide of the brachymemma that for example has the CBDAKD01 amino acid sequence of polypeptide, but lacked the one section continuous residue that comprises aminoterminal one section continuous residue or comprise C-terminal, or lacked two sections continuous residues, and one section comprises N-terminal, one section comprises C-terminal.The fragment that also preferably it is characterized in that structure or functional performance for example comprises the fragment of following structure: alpha-helix and alpha-helix form district, β-lamella and β-lamella and form district, corner and corner and form the district, curl and curl into district, hydrophilic area, hydrophobic region, α amphiphilic district, β amphiphilic district, flex region, surface and form district, substrate land and high antigenic index region.Other preferred fragments are bioactive fragments.Bioactive fragment is the active fragment of mediation CBDAKD01, comprises the fragment with similar activity or greater activity, or has low undesirable active fragment.Also be included in animal especially philtrum have antigenicity or immunogenic fragment.
Preferred all these polypeptide fragments have kept the biological activity of CBDAKD01, comprise antigenic activity.Given sequence and segmental variant also are parts of the present invention.Preferred variant is a conserved amino acid generation metathetical variant in the object of reference, and promptly the residue in the object of reference is had other amino-acid substitutions of similar characteristics.Typical this displacement comprises Ala, Val, the displacement between Leu and the Ile; Displacement between Ser and the Thr; Displacement between acidic residues Asp and the Glu; Displacement between Ash and the Gln; Displacement between alkaline residue Lys and the Arg; Or the displacement between aromatic residue Phe and the Tyr.Particularly preferably be wherein the variant of replacing, lacking or increase the combination of several, 5-10,1-5 or 1-2 amino acid or these modes.
CBDAKD01 polypeptide of the present invention can pass through prepared in any suitable way.These polypeptide comprise isolating naturally occurring polypeptide, the polypeptide of recombinant production, the synthetic polypeptide of producing, or the polypeptide of these method combinations produce.The method for preparing these polypeptide is widely known by the people in this area.Polynucleotide of the present invention
The present invention relates to the CBDAKD01 polynucleotide on the other hand.The CBDAKD01 polynucleotide comprise coding CBDAKD01 polypeptide and segmental isolating polynucleotide and polynucleotide closely-related with it.More specifically, CBDAKD01 polynucleotide of the present invention comprise the nucleotide sequence that contains sequence 1, encoding sequence 2 the CBDAKD01 polypeptide polynucleotide and have the polynucleotide of the particular sequence of sequence 1.The CBDAKD01 polynucleotide comprise that also the identity that contains with the nucleotide sequence total length of the CBDAKD01 polypeptide of encoding sequence 2 is at least the polynucleotide of 82% nucleotide sequence and contains the polynucleotide that identity with sequence 1 total length is at least 82% nucleotide sequence.Preferred identity is at least 90% polynucleotide, and especially preferred identity is at least 95% polynucleotide.It is highly preferred that identity is at least 97% polynucleotide, and more preferably identity is at least the polynucleotide of 98-99%, and most preferably identity is at least 99% polynucleotide.The CBDAKD01 polynucleotide also comprise its identity enough make its be used for increasing or as the condition of probe or mark under can with the nucleotide sequence of the nucleotide sequence hybridization of sequence 1.The present invention also provides and CBDAKD01 polynucleotide complementary polynucleotide.
Other albumen with Angiotensin and/or II/vasopressin receptor family on the CBDAKD01 structure of the present invention are relevant, and result's (sequence 1) of the cDNA order-checking of coding people CBDAKD01 has confirmed this point in the table 1.The cDNA sequence of sequence 1 contains an open reading frame (Nucleotide 47-1588) of 514 amino acid whose polypeptide of encoding sequence 2.The identity of the aminoacid sequence of table 2 (sequence 2) 485 amino-acid residues and rat serum angiotensin/II/vasopressin receptor (AII/AVP) (N.Ruiz-Opazo, et al., Nature, Med.1:074-1081,1995) is approximately 30% (FASTA).The identity of the nucleotide sequence of table 1 (sequence 1) 489 nucleotide residues and rat serum angiotensin/II/vasopressin receptor (AII/AVP) (N.Ruiz-Opazo, et al., Nature, Med.1:074-1081,1995) is approximately 57.9% (FASTA).Therefore, expect that CBDAKD01 polypeptide of the present invention and polynucleotide have and its homeopeptide and the similar biological function/characteristic of polynucleotide, its purposes is apparent to those skilled in the art.
Table 1 a
Figure 9881389200131
Figure 9881389200141
aPeople CBDAKD01 nucleotide sequence (sequence 1).
Table 2 b
Figure 9881389200142
bPeople CBDAKD01 aminoacid sequence (sequence 2).
The encode polynucleotide of CBDAKD01 of the present invention can obtain from the cDNA library by standard clone and screening, this library uses expressed sequence mark (EST) to analyze (Adams that derives by the mRNA from human umbilical cord blood cell, M.K., et al., Science (1991) 252:1651-1656; Adams, M.D., et al., Nature, (1992) 355:632-634; Adams, M.D., et al., Nature (1995) 377 Supp:3-174).Polynucleotide of the present invention also can maybe can use well-known commercially available technology synthetic from natural origin such as genome dna library.
The nucleotide sequence of the CBDAKD01 polypeptide of encoding sequence 2 can be equal to the polypeptid coding sequence (the Nucleotide 47-1588 of sequence 1) of table 1, perhaps can be the sequence of the polypeptide of encoding sequence 2 because genetic code Feng Yu (degeneracy) forms.
When polynucleotide of the present invention were used for recombinant production CBDAKD01 polypeptide, these polynucleotide can comprise mature polypeptide or its segmental encoding sequence itself; The sequence of mature polypeptide or fragment coding sequence and other encoding sequence frame endomixis, for example leading or secretion sequence, preceding former or preceding former protein sequence or other fusion polypeptide encoding sequence partly of other encoding sequences.For example can encode and help the flag sequence of fusion polypeptide purifying.In this certain preferred embodiments on the one hand of the present invention, flag sequence is 6-Histidine peptide or HA mark, the former with the pQE carrier provide (Qiagen, Inc.), relevant discussion is referring to Gentz et al., Proc Natl Acad Sci USA (1989) 86:821-824.That polynucleotide also can contain is noncoding 5 ' or 3 ' sequence, as transcribe, non-translated sequence, shear and polyadenylation signal the sequence of ribosome bind site and stable mRNA.
Further preferred embodiment is the polynucleotide of coding CBDAKD01 variant, described variant comprises the CBDAKD01 polypeptid acid sequence (sequence 2) of table 2, wherein replaces, lacks or increase several, 5-10,1-5,1-3,1-2 or 1 amino-acid residue or the combination of these modes.
The invention still further relates to polynucleotide with above-mentioned sequence hybridization.The invention particularly relates under stringent condition polynucleotide with above-mentioned multi-nucleotide hybrid.In this article, " stringent condition " expression has only identity between the sequence to be at least 80%, preferably at least 90%, more preferably at least 95%, most preferably could to hybridize during 97-99%.
Polynucleotide of the present invention are identical or enough identical with sequence 1 or its segmental nucleotide sequence, therefore can be as the hybridization probe of cDNA and genomic dna, to separate full-length cDNA and the genomic clone of coding CBDAKD01, also can separate the cDNA and the genomic clone that have other genes (homologue and the straight gene that comprise the species of coding except that the people) of high sequence similarity with the CBDAKD01 gene to homologue.These hybridization techniques are well known by persons skilled in the art.The identity of general these nucleotide sequences and object of reference sequence is 80%, and preferred identity is 90%, and more preferably identity is 95%.Probe generally comprises at least 15 Nucleotide.Preferred these probes comprise at least 30 Nucleotide or at least 50 Nucleotide.Particularly preferred probe is 30 to 50 Nucleotide.
In one embodiment, the polynucleotide that obtain coding CBDAKD01 polypeptide comprise that the homologue of the species except that the people or straight method to homologue may further comprise the steps: apparatus has sequence 1 or its segmental label probe to screen suitable library under tight hybridization conditions; Separate the full-length cDNA and the genomic clone that contain described polynucleotide sequence.Therefore, on the other hand, CBDAKD01 polynucleotide of the present invention also comprise following nucleotide sequence, and its nucleotides sequence is listed under the stringent condition hybridizes with nucleotide sequence or its fragment of sequence 1.The CBDAKD01 polypeptide also comprises following polypeptide, and its aminoacid sequence is nucleotide sequence coded by what obtain under the above-mentioned hybridization conditions.These hybridization techniques are known for those skilled in the art.Tight hybridization conditions as mentioned above, perhaps be following condition in other words: 42C is incubated overnight in following solution, then about 65 ℃ with 0.1 * SSC washing filter membrane, described solution comprises 50% methane amide, 5 * SSC (150 mM NaCl, 15mM trisodium citrate), 50mM sodium phosphate (pH7.6), 5 * Denhardt solution, the salmon sperm DNA that 10% dextran sulfate and 20 μ g/ml sex change are sheared.
Polynucleotide of the present invention and polypeptide can be used as research reagent and material, are used for seeking animal and human's class treatment of diseases and diagnostic method.Carrier, host cell, expression
The present invention also relates to contain the carrier of one or more polynucleotide of the present invention, contain the host cell of carrier of the present invention and produce polypeptide of the present invention by recombinant technology through genetically engineered.Utilization is derived from the RNA of DNA construct of the present invention, and cell free translation system can be used to produce these albumen.
For carrying out recombinant production, host cell can be through genetically engineered expression system or its part that contains polynucleotide of the present invention.Polynucleotide are imported host cell can be realized by the method described in the multiple standards laboratory manual, described laboratory manual such as Davis et al., " molecular biology basic skills " (1986) and Sambrook et al., " molecular cloning: laboratory manual " the 2nd edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989), described method comprises calcium phosphate transfection, the transfection of DEAE-dextran mediation, transposition, microinjection, the transfection of cationic lipid mediation, electroporation, transduction, friction transforms, particle bombardment or infection.
The representative example of suitable host comprises bacterial cell, as suis, staphylococcus, intestinal bacteria, streptomycete and bacillus subtilis cell; The fungal cell is as yeast cell and aspergillus cell; Insect cell is as fruit bat S2 and fall army worm Sf9 cell; Zooblast is as CHO, COS, HeLa, C127,3T3, BHK, HEK 293 and Bowes melanoma cell; And vegetable cell.
Can use multiple expression system.These expression systems comprise karyomit(e), episome and viral deutero-system etc., for example derived from the carrier of bacterial plasmid, bacteriophage, transposon, yeast episome, insertion element, yeast chromosomal element, virus, with carrier derived from the said elements combination, described virus comprises baculovirus, papovavirus such as SV40, vaccinia virus, adenovirus, fowlpox virus, Pseudorabies virus and retrovirus, and the combination carrier is as the carrier derived from plasmid and bacteriophage genetic elements, clay and phagemid.Expression system can contain the control region of regulating and starting expression.In general, any system or carrier that is suitable for keeping in the host, breed or expresses polynucleotide all can use.Suitable nucleotide sequence can be inserted in the expression system by multiple known routine techniques, and described technology can be referring to for example Sambtook et al., " molecular cloning: laboratory manual " the 2nd edition (the same).
For the albumen of translating can be secreted in the endoplasmic, periplasmic space or born of the same parents' external environment, can on required polypeptide, add suitable secretion signal.These signals can be that polypeptide is endogenous, also can be the external source secretion signals.
Be used for shaker test if the CBDAKD01 polypeptide will be expressed, general preferred polypeptide generates at cell surface.In this case, cell can be gathered in the crops before being used for shaker test.If the CBDAKD01 polypeptide is secreted in the substratum, can reclaim substratum to reclaim and purified polypeptide; If generate in cell, cell is at first wanted cracking, reclaims polypeptide then.The CBDAKD01 polypeptide can reclaim and purifying from the reconstitution cell culture by currently known methods, described method comprises ammonium sulfate or ethanol sedimentation, the acid extracting, negatively charged ion or cation-exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatograph, affinity chromatography, hydroxylapatite chromatography and lectin chromatogram.Most preferably high performance liquid chromatography is used for purifying.If polypeptide sex change in the isolated or purified process, the proteic method of known refolding can be used for recovering activity conformation.Diagnostic test
The invention still further relates to of the application of CBDAKD01 polypeptide as diagnostic reagent.Detect the CBDAKD01 transgenation form relevant with dysfunction a kind of diagnostic tool can be provided, this can increase or limit a kind of expression deficiency owing to CBDAKD01, express excessive or the disease of abnormal expression generation or the diagnosis of disease susceptibility.The individuality that carries sudden change in the CBDAKD01 gene can detect on dna level by multiple technologies.
Diagnosis nucleic acid can be available from experimenter's cell, as blood, urine, saliva, biopsy sample or postmortem material.Genomic dna can directly be used for detecting, or uses PCR or other amplification technique enzymatic amplifications before analyzing.RNA or cDNA can use in a similar manner.Disappearance can detect by the size variation of comparing amplified production with normal genotype with inserting.Point mutation can be by differentiating the CBDAKD01 nucleotide sequence hybridization of DNA amplification and mark.Pi Pei sequence can be by difference on RNA enzymic digestion or the melting temp and the double-stranded differentiation of mispairing fully.Dna sequence dna difference also can add or not detect with electrophoretic mobility on the gel of denaturing agent or direct dna sequencing by dna fragmentation.Can be referring to for example Myers et al., Science (1985) 230:1242.Also can show in the sequence variation of specific position by nuclease protection test such as RNA enzyme and S1 protection method or chemical chop method.Can be referring to Cotton et al., PNAS USA (1985) 85:4397-4401.In another embodiment, can make up and contain CBDAKD01 nucleotide sequence or its segmental oligonucleotide probe array (array), so that carry out effective screening of for example transgenation.The array technique method is widely known by the people, and suitability is extensive, can be used to solve the various problems in the molecular genetics, comprises genetic expression, genetic linkage and hereditary variability.(referring to for example M.Chee et al., Science, Vol.274, pp610-613 (1996).
Diagnostic test detects the sudden change of CBDAKD01 gene by utilizing aforesaid method, diagnosis is provided or has determined method to the susceptibility of hypertension, heart trouble, cancer and ephrosis.
In addition, hypertension, heart trouble, cancer and ephrosis can be diagnosed by the following method: determine to reduce or increase from the horizontal abnormality of CBDAKD01 polypeptide in experimenter's sample or CBDAKD01 mRNA.Expressing to reduce or increase to use the known any method in the quantitative field of Nucleotide to determine that on rna level described method is PCR, RT-PCR, RNA enzyme protection, RNA trace and other hybridizing methods for example.Can be used for determining that the experimental technique from protein level in host's the sample such as CBDAKD01 polypeptide level is well known to those skilled in the art.These test methods comprise that radioimmunoassay, competition are in conjunction with test, western blot analysis and ELISA test.
Therefore, on the other hand, the present invention relates to determine the Sickledex of disease or disease susceptibility, described disease is hypertension, heart trouble, cancer and ephrosis particularly, described medicine box comprises: (a) CBDAKD01 polynucleotide, the nucleotide sequence of preferred sequence 1, or its fragment; (b) with (a) in Nucleotide complementary nucleotide sequence; (c) CBDAKD01 polypeptide, the polypeptide of preferred sequence 2, or its fragment; (d) antibody of CBDAKD01 polypeptide, the antibody of the polypeptide of preferred sequence 2.Should be appreciated that in this medicine box, (a), (b), (c) or (d) can comprise a kind of main component (substantial component).The karyomit(e) test
Nucleotide sequence of the present invention is identified also valuable to karyomit(e).This sequence can special target one by one the specific position on the Autosome and can with its hybridization.The mapping of karyomit(e) correlated series is the important the first step that the gene with these sequences and disease-related connects according to the present invention.In case sequence has been positioned in concrete chromosome position, the physical location of this sequence on karyomit(e) can connect with the genetic map data.These data can be referring to for example V.McKusick, " human Mendelian inheritance " (by the online acquisition of Johns Hopkins University Welch Medical Library).Being positioned at the gene of same chromosomal region and the relation between the disease can pass through linkage analysis (the common heredity of the adjacent gene of physics) and determine.
The difference of cDNA or genome sequence also can be determined in the influenced and normal individual.If observe a certain sudden change in some or all affected individuals, and do not observe this sudden change in normal individual, then this sudden change may be the cause of disease of disease.The CBDAKD01 gene has been positioned in the q21.3 on the X chromosome.Antibody
Polypeptide of the present invention or its fragment or analogue or the cell of expressing them also can be used as immunogen, generate the specific antibody of CBDAKD01 polypeptide immune." immunologic opsonin " represents that in the art avidity to polypeptide of the present invention is obviously greater than the avidity to other related polypeptides of this area.
Can be at the antibody of CBDAKD01 polypeptide by ordinary method to the preferred inhuman animals administer polypeptide of animal or contain epi-position fragment, analogue or cell and obtain.In order to prepare monoclonal antibody, can use any technology of the antibody that the generation of continuous cell line culture is provided.Example comprises hybridoma technology (Kohler, G.and Milstein, C., Nature (1975) 256:495-497), three knurls (trioma) technology, human B cell hybridoma technology (Kozbor et al., Immunology Today (1983) 4:72) and EBV hybridoma technology (Cole etal., " monoclonal antibody and cancer therapy ", pp77-96, Alan R.Liss, Inc., 1985).
After also can improving, the technology of manufacture order chain antibody (United States Patent (USP) 4,946,778) is used for the single-chain antibody of production polypeptide of the present invention.Transgenic mice or other organisms comprise that other Mammalss also can be used for expressing humanized antibody.
Above-mentioned antibody also can be used for separating or identify the clone of the described polypeptide of expression or the polypeptide of purifying affinity chromatography technology preparation.
The antibody of anti-CBDAKD01 polypeptide also can be used for treating hypertension, heart trouble, cancer and ephrosis etc.Vaccine
Other aspects of the present invention relate to the method for induce immune response in Mammals; comprise with CBDAKD01 polypeptide or its suitable fragment immunization Mammals; produce antibody and/or t cell response, thereby protect this animal to avoid diseases such as hypertension, heart trouble, cancer and ephrosis.Other aspects of the present invention also relate to the method for induce immune response in Mammals, comprise that administration instructs the carrier of CBDAKD01 polypeptide expression in vivo to come administration CBDAKD01 polypeptide, so that induce immune response, generate antibody and watch for animals and avoid disease.
Other aspects of the present invention relate to immunogen/vaccine preparation (composition), when being introduced into mammalian hosts, can induce the immunne response at the CBDAKD01 polypeptide, and wherein said composition contains CBDAKD01 polypeptide or CBDAKD01 gene.This vaccine preparation can also contain suitable carriers.Because the CBDAKD01 polypeptide can be degraded under one's belt, preferably by parenteral route administration (comprising injections such as subcutaneous, muscle, intravenously, intracutaneous).The preparation that is fit to parenteral admin comprises moisture and water-free aseptic parenteral solution, wherein can contain antioxidant, buffer reagent, fungistat and make preparation and the isoosmotic solute of receptor's blood; Moisture and water-free sterile suspensions wherein can contain suspension agent or thickening material.Said preparation can be single agent or multi-agent vessel form, and for example sealed ampoule and bottle also can be kept under the lyophilisation condition, only need add sterile liquid carrier before use and get final product.Vaccine preparation also can contain the immunogenic adjuvant system of enhancing preparation, as oil-in-water system or other system known in the art.Dosage depends on the specific activity of vaccine, can determine easily by routine test.Shaker test
CBDAKD01 polypeptide of the present invention can be used for screening and activating (agonist) or suppress the method for (antagonist or be called inhibitor) CBDAKD01 polypeptide activated compound of the present invention.Therefore polypeptide of the present invention also can be used for estimating from cell, acellular prepared product, chemical library and natural product mixture and identify agonist or antagonist.These agonists or antagonist can be the natural of polypeptide of the present invention or the substrate of modifying, part, acceptor, enzyme etc.; Or the structure of polypeptide of the present invention or functional analogue thing.Can be referring to Coligan et al., Current Protocol inImmunology 1 (2): Chapter 5 (1991).
The CBDAKD01 polypeptide has different physiological roles, comprises multiple pathologic effect.Therefore, be necessary to seek compound and the medicine that stimulates the CBDAKD01 polypeptide or suppress CBDAKD01 polypeptide function.Generally, agonist can be used for treatment of diseases and prevention purposes such as hypertension, heart trouble, cancer and ephrosis.Antagonist can be used for the multiple treatment and the prevention purpose of diseases such as hypertension, heart trouble, cancer and ephrosis.
Generally, these screening methods comprise the suitable cell that uses expression CBDAKD01 polypeptide or CBDAKD01 polypeptide of the present invention is responded.These cells comprise from Mammals, yeast, fruit bat or colibacillary cell.To express the cell cytolemma of express polypeptide (or contain) of CBDAKD01 polypeptide or the cell that the CBDAKD01 polypeptide responds contact the stimulation or the inhibition of observation combination or functional response with testing compound.The cell of more contacted candidate compound and the active ability of isocellular CBDAKD01 of contacted candidate compound not.
This test can be tested the combination of candidate compound simply, wherein passes through directly or indirectly and candidate compound bonded marker detection with the adhesion of cell with CBDAKD01 polypeptide, or by comprising that the test of competing with mark competition thing detects.And whether these tests can be tested candidate compound can produce the signal that the CBDAKD01 polypeptide activate to produce, and uses the detection system of the cell that is suitable for having the CBDAKD01 polypeptide.Activation inhibitor test when known agonist exists usually, observe candidate compound exist to agonist activated shadow to.
In addition, this test can may further comprise the steps simply: mix candidate compound and the solution that contains the CBDAKD01 polypeptide, form mixture, measure CBDAKD01 activity in the mixture, relatively the CBDAKD01 activity of mixture and standard substance.
CBDAKD01 cDNA, albumen and proteic antibody also can be used for designing the test that detects the influence that CBDAKD01 mRNA and albumen generate in the compound pair cell that adds.For example, can design ELISA, use mono-clonal and polyclonal antibody to measure secretion or cell bonded CBDAKD01 protein level by standard method known in the art, this can be used for seeking from suitable engineered cell or tissue and can suppress or increase the material (also can be called antagonist or agonist) that CBDAKD01 generates.
CBDAKD01 albumen can be used for identifying the film combination or the solvable acceptor of existence, uses standard receptors bind technology known in the art.These include but not limited to part combination and crosslinked test, wherein CBDAKD01 is with radio isotope (125I) mark, chemically modified (as biotinylation), or be fit to detect or the peptide sequence of purifying merges, with infer be subjected to body source (cell, cytolemma, cell conditioned medium, tissue extract, body fluid) incubation.Additive method comprises the biophysics technology, as surperficial kytoplasm resonance and spectroscopy.Except being used for purifying and the clone's acceptor, these also can be used for identifying CBDAKD01 agonist and antagonist in conjunction with test, they and the combine competition of CBDAKD01 to the acceptor of existence.The standard method of carrying out shaker test is well known in the art.
The example of possible CBDAKD01 polypeptide antagonist comprises the antibody of CBDAKD01 polypeptide, perhaps be in some cases and closely-related oligonucleotide or albumen such as the part of CBDAKD01 polypeptide, substrate, acceptor, enzyme, for example fragment of part, substrate, acceptor, enzyme etc.; Perhaps be to combine with polypeptide of the present invention but the small molecules of initiating response not, thereby suppress the activity of polypeptide.
Therefore, the present invention relates to a kind of screening medicine box on the other hand, is used for identifying agonist, antagonist, part, acceptor, substrate, enzyme of CBDAKD01 polypeptide etc.; Or the compound of reduction or the generation of increase CBDAKD01 polypeptide, this medicine box comprises: (a) CBDAKD01 polypeptide, the polypeptide of preferred sequence 2; (b) reconstitution cell of the polypeptide of expression CBDAKD01 polypeptide preferred sequence 2; (c) cytolemma of the polypeptide of expression CBDAKD01 polypeptide preferred sequence 2; (d) antibody of CBDAKD01 polypeptide, the antibody of the polypeptide of preferred sequence 2.Should be appreciated that in this medicine box, (a), (b), (c) or (d) can comprise a kind of main component.Prevention and methods of treatment
The invention provides the methods of treatment with the too high or too low diseases associated of CBDAKD01 polypeptide active, described diseases, such as hypertension, heart trouble, cancer and ephrosis etc.
If the CBDAKD01 polypeptide active is too high, several method can be arranged.A kind of method comprises to the above-mentioned inhibitor compound of experimenter's administration (antagonist) and pharmaceutically acceptable carrier, its consumption can suppress the function of CBDAKD01 polypeptide, for example pass through the combination of block ligand, substrate, acceptor, enzyme etc., or by the inhibition second signal, thereby abnormal symptom alleviated.Administration can be competed the CBDAKD01 polypeptide of the soluble form of binding partner, substrate, enzyme, acceptor etc. with endogenous CBDAKD01 polypeptide in the another kind method.The representative instance of this competition thing comprises the fragment of CBDAKD01 polypeptide.
In another approach, can use the expression of gene of expressing the endogenous CBDAKD01 polypeptide of interrupter technique inhibition coding.Known this technology comprises the use antisense sequences, can be inner that produce or individually dosed.Can be referring to for example O ' Connor, J Neurochem (1991) 56:560, " oligodeoxynucleotide of genetic expression antisense inhibitor " CRC Press, Boca Raton, FL (1988).Perhaps can use the oligonucleotide that forms triple helical with gene.Referring to for example Lee, et al., Nucleic Acid Res (1979) 6:3073; Cooney et al., Science (1988) 241:456; Dervan et al., Scicence (1991) 251:1360.Can these oligomer of administration or express relevant oligomer in vivo.
In order to treat CBDAKD01 and the insufficient abnormal symptom of its activity expression, there is several method to use.A kind of method comprises the compound to the activation CBDAKD01 of experimenter's drug treatment significant quantity polypeptide, and promptly above-mentioned agonist in conjunction with the pharmaceutically useful carrier of administration, thereby is alleviated abnormal symptom.Perhaps, can use gene therapy to realize experimenter's relevant cell endogenous production CBDAKD01.For example, polynucleotide of the present invention can be expressed with the replication defect type retroviral vector through engineered as above-mentioned.Separate then in retrovirus expression construct and the packing cell of importing, contain the RNA of code book invention polypeptide in the carrier, thereby packing cell can be produced the infectious virus particle that contains goal gene with the retroviral plasmid vector transduction.Can these produce cells to experimenter's administration, so that engineered cells and express polypeptide in vivo in the body.The relevant discussion of gene therapy can be referring to the 20th chapter, " gene therapy and other methods of treatment (and reference wherein); " human molecular genetics ", T Strachan and A P Read, BIOSScientific Publishers Ltd (1996) based on molecular genetics.Another kind method is the CBDAKD01 polypeptide of drug treatment significant quantity, in conjunction with the suitable pharmaceutical carrier of administration.Preparation and administration
Peptides such as the soluble form of CBDAKD01 polypeptide and peptide or micromolecular agonist and antagonist can with suitable pharmaceutical carrier combination formula.These preparations comprise polypeptide or compound and the pharmaceutically acceptable carrier or the vehicle for the treatment of significant quantity.These carriers include but not limited to salt solution, buffer saline, glucose, water, glycerine, ethanol and its combination.Preparation should be fit to administering mode, and this is well known to those skilled in the art.The invention still further relates to cartridge bag and medicine box,, one or more above-mentioned composition compositions of the present invention are housed comprising one or more containers.
Polypeptide of the present invention and other compounds can use separately, or unite use with other compounds as the treatment compound.
The preferred whole body administering mode of medicinal compositions comprises injection, generally is intravenous injection.Also can use other injecting pathways such as subcutaneous, muscle or intraperitoneal approach.The additive method of whole body administration comprises that use cholate or permeate agents such as fusidinic acid or other tensio-active agents are through mucous membrane, percutaneous dosing.In addition, if be mixed with enteron aisle or capsule, also can be taken orally.These compounds also can be with form surface and/or topicals such as ointment, paste, gels.
The required dosage scope depends on selection, route of administration, preparation nature, the character of experimenter's symptom and attending doctor's the judgement of peptide.Suitable dosage ranges is 0.1 to 100 μ g/kg.In view of the compound that can obtain has multiple, different route of administration different effects is arranged, the variation range of required dosage is very big to be predictable.For example, the expection oral administration is big than the intravenous injection required dosage.The variation of these dosage levels can be used the standard test method adjustment that is used for optimizing, and this is well known in the art.
Treatment also can endogenous generation in subject with polypeptide, and this form of therapy is commonly referred to " gene therapy ".Therefore, for example can use DNA or the RNA of polynucleotide, utilize retroviral plasmid vector engineered as the polypeptide of coding ex vivo from experimenter's cell.Then these cells are introduced in the subject.
All publications of being quoted in this specification sheets include but not limited to patent and patent application, and are all incorporated by reference, indicate incorporated by reference separately and individually as each publication.
Sequence table
(1) general information (ⅰ) applicant: Zhang Qinghua, Shen Yu, Mao Mao, Gu Baiwei (ⅱ) denomination of invention: human angiotensin II/II/vasopressin receptor (AII/AVP) sample base
Cause (CBDAKD01) is sequence number (ⅲ): 2 (ⅳ) address:
(A) contact person: Ratner ﹠amp; Prestia
(B) street: P.O.Box 980
(C) city: Valley Forge
(D) state: PA
(E) country: USA
(F) postcode: 19482 (ⅴ) computer-reader form:
(A) media type: floppy disk
(B) computer: IBM compatibility
(C) operating system: DOS
(D) software: the current request for data of FastSEQ for Windows Version 2.0 (ⅵ):
(A) application number:
(B) applying date:
(C) classification: (ⅶ) request for data formerly:
(A) application number:
(B) applying date: (ⅷ) lawyer/proxy's information:
(A) name: Prestia, Paul F
(B) registration number: 23,031
(C) reference/file number: GP-70382 (ⅸ) telecommunication information:
(A) phone: 610/407-0700
(B) fax: 610/407-0701
(C) fax: 846169
(2) sequence 1 information: (ⅰ) sequence signature:
(A) length: 2218 base pairs
(B) type: nucleic acid
(C) chain: strand
( D ) ： ( ⅱ ) ：cDNA ( ⅹⅰ ) ：1：CAGGGCAGCC TTCAGTCTGA TTCAGGAGAA CGAGGTCCTC TTCACCATGT GCTTCATCCC 60CCTGGTCTGC TGGATCGTGT GCACTGGACT GAAACAGCAG ATGGAGAGTG GCAAGAGCCT 120TGCCCAGACA TCCAAGACCT CCACCGCGGT GTACGTCTTC TTCCTTTCCA GTTTGCTGCA 180GCCCCGGGGA GGGAGCCAGG AGCACGGCCT CTGCGCCCAC CTCTGGGGGC TCTGCTCTTT 240GGCTGCAGAT GGAATCTGGA ACCAGAAAAT CCTGTTTGAA GAGTCCGACC TCAGGAATCA 300TGGACTGCAG AAGGCGGATG TGTCTGCTTT CCTGAGGATG AACCTGTTCC AAAAGGAAGT 360GGACTGCGAG AAGTTCTACA GCTTCATCCA CATGACTTTC CAGGAGTTCT TTGCCGCCAT 420GTACTACCTG CTGGAAGAGG AAAAGGAAGG AAGGACGAAC GTTCCAGGGA GTCGTTTGAA 480GCTTCCCAGC CGAGACGTGA CAGTCCTTCT GGAAAACTAT GGCAAATTCG AAAAGGGGTA 540TTTGATTTTT GTTGTACGTT TCCTCTTTGG CCTGGTAAAC CAGGAGAGGA CCTCCTACTT 600GGAGAAGAAA TTAAGTTGCA TGATCTCTCA GCAAATCAGG CTGGAGCTGC TGAAATGGAT 660TGAAGTGAAA GCCAA GCTA AAAAGCTGCA TGATCAGCCC AGCCAGCTGG AATTGTTCTA 720CTGTTTGTAC GAGATGCAGG AGGAGGACTT CGTGCAAAGG GCCATGGACT ATTTCCCCAA 780GATTGAGATC AATCTCTCCA CCAGAATGGA CCACATGGTT TCCTCCTTTT GCATTGAGAA 840CTGTCATCGG GTGGAGTCAC TGTCCCTGGG GTTTCTCCAT AACATGCCCA AGGAGGAAGA 900GGAGGAGGAA AAGGAAGGCC GACACCTTGA TATGGTGCAG TGTGTCCTCC CAAGCTCCTC 960TCATGCTGCC TGTTCTCATG GGTTGGGGCG CTGTGGCCTC TCCCATGAGT GCTGCTTCGA 1020CATCTCCTTG GTCCTCAGCA GCAACCAGAA GCTGGTGGAG CTGGACCTGA GTGACAACGC 1080CCTCGGTGAC TTCGGAATCA GACTTCTGTG TGTGGGACTG AAGCACCTGT TGTGCAATCT 1140GAAGAAGCTC TGGTTGGTGA ATTCTGCCTT ACGTCAGTCT GTTGTTCAGC TTTGTCCTCG 1200GTACTCAGCA CTAATCAGAA TCTCACGCAC CTTTACTGCG AGGCAACACT CTCGGAGACA 1260AGGGATCAAA CTACTCTGTG AGGGACTCTT GCACCCCGAC TGcAAGCTTC AGGTGTTGGA 1320ATTAGACAAC TGCAACCTCA CGTCACACTG CTGCTGGGAT CTTTCCACAC TTCTGACCTC 1380CAGCCAGAGC CTGCGAAAGC TGAGCCTGGG CAACAATGAC CTGGGCGACC TGGGGGTCAT 1440GATGTTCTGT GAAGTGCTGA AACAGCAGAG CTGCCTCCTG CAGAACCTGG GGTTGTCTGA 1500AATGTATTTC AATTATGAGA CAAAAAGTGC GTTAGAAACA CTTCAAGAAG AAAAGCCTGA 1560GCTGACCGTC GTCTTTGAGC CTTCTTGGTA GGAGTGGAAA CGGGGCTGCC AGACGCCAGT 1620GTTCTCCGGT CCCTCCAGCT GGGGGCCCTC AGGTGGAGAG AGCTGCGATC CATCCAGGCC 1680AAGACCACAG CTCTGTGATC CTTCCGGTGG AGTGTCGGAG AAGAGAGCTT GCCGACGATG 1740CCTTCCTGTG CAGAGCTTGG GCATCTCCTT TACGCCAGGG TGAGGAAGAC ACCAGGACAA 1800TGACAGCATC GGGTGTTGTT GTCATCACAG CGCCTCAGTT AGAGGATGTT CCTCTGGTGA 1860CCTCATGTAA TTAGCTCATT CAATAAAGCA CTTTCTTTAT TTTTCTCTTC TCTGTCTAAC 1920CTTCTTTTTC CTATCTTTTT TTCTTCTTTG TTCTGTTTAC TTTTGCTCAT ATCATCATTC 1980CCGCTATCTT TCTATTAACT GACCATAACA CAGAACTAGT TGACTATATA TTATGTTGAA 2040ATTTTATGGC AGCTATTTAT TTATTTAAAT TTTTTGTAAT AGTTTTGTTT TCTAATAAGA 2100AAAATCCATG CTTTTTGTAG CTGGTTGAAA ATTCAGGAAT ATGTAAAACT TTTTGGTATT 2160TAATTAAATT GATTCCTTTT CTTAATTTTA AAAAAAAAAA AAAAAAAAAA AAAAAAAA 2218
(2) sequence 2 information: (ⅰ) sequence signature:
(A) length: 514 amino acid
(B) type: amino acid
(C) chain: strand
( D ) ： ( ⅱ ) ： ( ⅹⅰ ) ：2：Met Cys Phe Ile Pro Leu Val Cys Trp Ile Val Cys Thr Gly Leu Lys1 5 10 15Gln Gln Met Glu Ser Gly Lys Ser Leu Ala Gln Thr Ser Lys Thr Ser 20 25 30Thr Ala Val Tyr Val Phe Phe Leu Ser Ser Leu Leu Gln Pro Arg Gly 35 40 45Gly Ser Gln Glu His Gly Leu Cys Ala His Leu Trp Gly Leu Cys Ser 50 55 60Leu Ala Ala Asp Gly Ile Trp Asn Gln Lys Ile Leu Phe Glu Glu Ser65 70 75 80Asp Leu Arg Asn His Gly Leu Gln Lys Ala Asp Val Ser Ala Phe Leu 85 90 95Arg Met Asn Leu Phe Gln Lys Glu Val Asp Cys Glu Lys Phe Tyr Ser 100 105 110Phe Ile His Met Thr Phe Gln Glu Phe Phe Ala Ala Met Tyr Tyr Leu 115 120 125Leu Glu Glu Glu Lys Glu Gly Arg Thr Asn Val Pro Gly Ser Arg Leu 130 135 140Lys Leu Pro Ser Arg Asp Val Thr Val Leu Leu Glu Asn Tyr Gly Lys145 150 155 160Phe Glu Lys Gly Tyr Leu Ile Phe Val Val Arg Phe Leu Phe Gly Leu 165 170 175 Val Asn Gln Glu Arg Thr Ser Tyr Leu Glu Lys Lys Leu Ser Cys Met 180 185 190Ile Ser Gln Gln Ile Arg Leu Glu Leu Leu Lys Trp Ile Glu Val Lys 195 200 205Ala Lys Ala Lys Lys Leu His Asp Gln Pro Ser Gln Leu Glu Leu Phe 210 215 220Tyr Cys Leu Tyr Glu Met Gln Glu Glu Asp Phe Val Gln Arg Ala Met225 230 235 240Asp Tyr Phe Pro Lys Ile Glu Ile Asn Leu Ser Thr Arg Met Asp His 245 250 255Met Val Ser Ser Phe Cys Ile Glu Asn Cys His Arg Val Glu Ser Leu 260 265 270Ser Leu Gly Phe Leu His Asn Met Pro Lys Glu Glu Glu Glu Glu Glu 275 280 285Lys Glu Gly Arg His Leu Asp Met Val Gln Cys Val Leu Pro Ser Ser 290 295 300Ser His Ala Ala Cys Ser His Gly Leu Gly Arg Cys Gly Leu Ser His305 310 315 320Glu Cys Cys Phe Asp Ile Ser Leu Val Leu Ser Ser Asn Gln Lys Leu 325 330 335Val Glu Leu Asp Leu Ser Asp Asn Ala Leu Gly Asp Phe Gly Ile Arg 340 345 350Leu Leu Cys Val Gly Leu Lys His Leu Leu Cys Asn Leu Lys Lys Leu 355 360 365Trp Leu Val Asn Ser Ala Leu Arg Gln Ser Val Val Gln Leu Cys Pro 370 375 380Arg Tyr Ser Ala Leu Ile Arg Ile Ser Arg Thr Phe Thr A la Arg Gln385 390 395 400His Ser Arg Arg Gln Gly Ile Lys Leu Leu Cys Glu Gly Leu Leu His 405 410 415Pro Asp Cys Lys Leu Gln Val Leu Glu Leu Asp Asn Cys Asn Leu Thr 420 425 430Ser His Cys Cys Trp Asp Leu Ser Thr Leu Leu Thr Ser Ser Gln Ser 435 440 445Leu Arg Lys Leu Ser Leu Gly Asn Asn Asp Leu Gly Asp Leu Gly Val 450 455 460Met Met Phe Cys Glu Val Leu Lys Gln Gln Ser Cys Leu Leu Gln Asn465 470 475 480Leu Gly Leu Ser Glu Met Tyr Phe Asn Tyr Glu Thr Lys Ser Ala Leu 485 490 495Glu Thr Leu Gln Glu Glu Lys Pro Glu Leu Thr Val Val Phe Glu Pro 500 505 510Ser Trp

Claims (19)

1. isolating polynucleotide contain identity with the nucleotide sequence total length of the CBDAKD01 polypeptide of encoding sequence 2 and are at least 80% nucleotide sequence; The complementary nucleotide sequence of perhaps described separation polynucleotide.
2. the polynucleotide of claim 1, wherein said polynucleotide comprise the nucleotide sequence of sequence 1 of the CBDAKD01 polypeptide of encoding sequence 2.
3. the polynucleotide of claim 1, wherein said polynucleotide comprise that the identity with the nucleotide sequence total length of sequence 1 is at least 80% nucleotide sequence.
4. the polynucleotide of claim 3, it is the polynucleotide of sequence 1.
5. the polynucleotide of claim 1, it is DNA or RNA.
6. contain a kind of DNA or RNA molecule of expression system, wherein said expression system can be produced the CBDAKD01 polypeptide in compatible host cell, and the identity of the polypeptide of its aminoacid sequence and sequence 2 is at least 80%.
7. the host cell that contains the expression system of claim 6.
8. produce the method for CBDAKD01 polypeptide, be included in the host who cultivates claim 7 under the condition that can produce described polypeptide, by reclaiming polypeptide in the culture.
9. preparation produces the method for the cell of CBDAKD01 polypeptide, comprises that the expression system with claim 6 transforms or transfection host cell, makes host cell can produce the CBDAKD01 polypeptide under suitable culture condition.
10.CBDAKD01 polypeptide contains identity with the aminoacid sequence total length of sequence 2 and is at least 80% aminoacid sequence.
11. the polypeptide of claim 10, it comprises the aminoacid sequence of sequence 2.
12. the immunologic opsonin antibody of the CBDAKD01 polypeptide of claim 10.
13. the experimenter's who treats the activity of the CBDAKD01 polypeptide that needs increase claim 10 or express method comprises:
(a) to the agonist of the described polypeptide of experimenter's drug treatment significant quantity; And/or
(b) provide isolating polynucleotide to the experimenter, the identity of the coding nucleotide sequence total length of the CBDAKD01 polypeptide of its nucleotide sequence and sequence 2 is at least 80%; Perhaps its nucleotide sequence and above-mentioned nucleotide sequence complementation, the form of polynucleotide can realize the expression in vivo of described polypeptide active.
14. the experimenter's who treats the activity of the CBDAKD01 polypeptide that needs inhibition claim 10 or express method comprises:
(a) to the antagonist of the described polypeptide of experimenter's drug treatment significant quantity; And/or
(b) suppress the nucleic acid molecule that the coding nucleotide sequence of described polypeptide is expressed to experimenter's administration; And/or
(c) compete the polypeptide of its part, substrate or acceptor to experimenter's drug treatment significant quantity with described polypeptide.
15. with the method for the susceptibility of the CBDAKD01 polypeptide expression of claim 10 or active diseases associated or disease, comprising among the diagnosis experimenter:
(a) determine whether there is sudden change in the coding nucleotide sequence of CBDAKD01 polypeptide described in described experimenter's genome; And/or
(b) analyze from whether having CBDAKD01 polypeptide expression or its expression amount in experimenter's the sample.
16. identify the method for the compound of the CBDAKD01 polypeptide that suppresses (antagonism) or activation claim 10, comprising:
(a) with candidate compound and the cell (or cytolemma of expression CBDAKD01 polypeptide) of expression CBDAKD01 polypeptide or the cells contacting that the CBDAKD01 polypeptide is responded; With
(b) stimulation or the inhibition of observation combination or functional response; Or the cell of more contacted candidate compound (or cytolemma) and the ability of the isocellular CBDAKD01 polypeptide active of contacted candidate compound not.
17. the agonist that the method for claim 16 is identified.
18. the antagonist that the method for claim 16 is identified.
19. its cytolemma of the recombinant host cell of the method for claim 9 preparation or expression CBDAKD01 polypeptide.
CN98813892.1A 1998-03-12 1998-03-12 Human angiotonin II/vasopressin receptor like gene Pending CN1286698A (en)

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