CN1318557A - Human testicular specific protein-10 gene encoded protein - Google Patents
Human testicular specific protein-10 gene encoded protein Download PDFInfo
- Publication number
- CN1318557A CN1318557A CN 01113505 CN01113505A CN1318557A CN 1318557 A CN1318557 A CN 1318557A CN 01113505 CN01113505 CN 01113505 CN 01113505 A CN01113505 A CN 01113505A CN 1318557 A CN1318557 A CN 1318557A
- Authority
- CN
- China
- Prior art keywords
- gene
- nyd
- protein
- reading frame
- open reading
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The present invention belongs to the field of human genome technology. The human testis specific protein-10 (NYD-SP10) gene has whole length cDNA sequence of 3382 bp, open reading frame sequence 1926 bp, 641 encoded amino acids and Genebank number of AF332192. The present invention utilizes NYD-SP10 gene clone to prepare fusion protein, utilizes the protein to immunize animal and to prepare monoclonal and polyclonal antibody, and utilizes gene clone in preparing texticular specific function gene expressing chip. The expressed protein may become gene medicine for treating related disease.
Description
The invention belongs to the Human genome technical group field.
TESTIS SPECIFIC PROTEIN 1's 0 gene of the present invention is that the contriver obtains with the method for gene chip screening in reproductive medicine Jiangsu Province key lab of Nanjing Medical University, at first prepare people's testicular function gene expression chip, by comparing embryo and grownup's testicular function expression of gene, acquisition differential expression clone, prove full-length cDNA through sequencing, be 3382bp; Its open reading frame is 1926bp, its 641 amino acid of encoding.Consult gene pool, this gene is new gene, at testis specific expression.Because this gene is the adult testis high expression level, therefore TESTIS SPECIFIC PROTEIN 1's 0 gene of finding may be relevant with testicualr development, can be prepared into fusion rotein with this gene clone, with this protein immunization rabbit, can obtain mono-clonal and polyclonal antibody, this gene also can be used for preparing the testicular function gene expression chip.Fusion rotein also may become the genomic medicine for the treatment of correlative diseases
Because TESTIS SPECIFIC PROTEIN 1's 0 gene still unmanned homologous gene in the Genbank database, the inventor all is initiative for the 26S Proteasome Structure and Function achievement in research of this gene.(National Center for BiotechnologyInformation NCBI) formally accepts the newcomer that this gene is Genbank, and accepting number is AF332192 in American National biotechnology information center.1?aggtgggaag?gcagttatga?cagttgagaa?gtagtagaag?acacggaagg?cacagaaggc 61?agacttcgct?cagcacaaag?aagaattttc?tgataaccat?actggcaaaa?tgaactgggc?121?tgccttcgga?gggtctgaat?tcttcatccc?agaaggcatt?cagatagatt?cgagatgccc?181?actaagcaga?aatatcacgg?aatggtacca?ttactatggc?attgcagtga?aagaaagctc?241?ccaatattat?gatgtgatgt?attccaagaa?aggagctgcc?tgggtgagtg?agacgggcaa?301?gaaagaagtg?agcaaacaga?cagtggcata?ttcaccccgg?tccaaactcg?gaacactgct?361?gccagaattt?cccaatgtca?aagatctaaa?tctgccagcc?agcctgcctg?aggagaaggt?421?ttctaccttt?attatgatgt?acagaacaca?ctgtcagaga?atactggaca?ctgtaataag?481?agccaacttt?gatgaggttc?aaagtttcct?tctgcacttt?tggcaaggaa?tgccgcccca?541?catgctgcct?gtgctgggct?cctccacggt?ggtgaacatt?gtcggcgtgt?gtgactccat?601?cctctacaaa?gctatctccg?gggtgctgat?gcccactgtg?ctgcaggcat?tacctgacag?661?cttaactcag?gtgattcgaa?agtttgccaa?gcaactggat?gagtggctaa?aagtggctct?721?ccacgacctc?ccagaaaact?tgcgaaacat?caagttcgaa?ttgtcgagaa?ggttctccca?781?aattctgaga?cggcaaacat?cactaaatca?tctctgccag?gcatctcgaa?cagtgatcca?841?cagtgcagac?atcacgttcc?aaatgctgga?agactggagg?aacgtggacc?tgaacagcat?901?caccaagcaa?accctttaca?ccatggaaga?ctctcgcgat?gagcaccgga?aactcatcac?961?ccaattatat?caggagtttg?accatctctt?ggaggagcag?tctcccatcg?agtcctacat1021?tgagtggctg?gataccatgg?ttgaccgctg?tgttgtgaag?gtggctgcca?agagacgagg1081?gtccttgaag?aaagtggccc?agcagttcct?cttgatgtgg?tcctgtttcg?gcacaagggt1141?gatccgggac?atgaccttgc?acagcgcccc?cagcttcggg?tcttttcacc?taattcactt1201?aatgtttgat?gactacgtgc?tctacctgtt?agaatctctg?cactgtcagg?agcgggccaa1261?tgagctcatg?cgagccatga?agggagaagg?aagcactgca?gaagtccgag?aagagatcat1321?cttgacagag?gctgccgcac?caaccccttc?accagtgcca?tcgttttctc?cagcaaaatc1381?tgccacatct?gtggaagtgc?cacctccctc?ttcccctgtt?agcaatcctt?cccctgagta1441?cactggcctc?agcactacag?gagcaatgca?ggcttacacg?tggtctctaa?catacacagt1501?gacgacggct?gctgggtccc?cagctgagaa?ctcccaacag?ctgccctgta?tgaggaacac1561?tcacgtgcct?tcttcctccg?tcacacacag?gataccagtt?tatccccaca?gagaggaaca1621?tggatacacg?ggaagctata?actatgggag?ctatggcaac?cagcatcctc?accccatgca1681?gagccagtat?ccggccctcc?ctcatgacac?agctatctct?gggccactcc?actatgcccc1741?ttaccacagg?agctctgcac?agtacccttt?taatagcccc?acttcccgga?tggaaccttg1801?tttgatgagc?agtactccca?gactgcatcc?taccccagtc?actccccgct?ggccagaggt1861?gccctcagcc?aacacgtgct?acacaaaccc?gtctgtgcat?tctgcgaggt?acggaaactc1921?tagtgacatg?tatacacctc?tgacaacgcg?caggaattct?gaatatgagc?acatgcaaca1981?ctttcctggc?tttgcttaca?tcaacggaga?ggcctctaca?ggatgggcta?aatgactgct2041?atcataggca?tccatattta?atattaataa?taataattaa?taataataat?aaacccaaca2101?cccatccccc?agaagacttt?atctctatac?attgtaactc?atgggctatt?cctaagtgcc2161?cattttccta?atgaacatga?ggatgggatc?aatgtgggat?gaataaactt?tagttcagaa2221?acaggactta?ctaaaagtca?gtgggactgg?gtttctgtag?ccaagccaga?cttgactgtt2281?tctgtagagc?actatctcgg?gcaggccatt?ctgtgccttt?tccctctgtt?ccatgacttt2341?gctttgtgtt?ggcaaccact?tctagtaagc?tactgatttt?cctgttgaca?aaatctcttt2401?agtcttgaag?gatggatact?ggagacagaa?tctggtttgt?gttcttggat?gggcacataa2461?tttaccaaga?gcattcacct?tgccatctgt?cttgtcattg?tactgtacaa?ggaacagccc2521?tcagacgtgt?tctgcacatc?ccttcttcct?ggtggtacca?tccctatttc?ctggagcacc2581?agggctaaat?ggggagctat?ctggaaactc?tagattttct?gtcataccca?catctgtcac2641?agtacctgca?ttgtcttgga?atgtaagcac?tgtcttgagg?gaaggaagag?gtctgttctg2701?tattgcctta?agttgattga?ggtttgtagg?agactggttc?ttctacatac?aaggatttgt2761?cttaagtttg?cacaatggct?agtgtcagca?aaaggcagga?gagggttttt?gttttttttt2821?taagttctat?gagaatgtgg?atttatggca?ttgagtatca?cactcagctc?tgctgtgtta2881?actttgtgaa?actggatgga?acaaacttta?acttaccaag?caccaagtgt?gaaagtgact2941?ttcacggttc?cttcataaaa?ctataataat?atccgacact?ttgatagaaa?aaaattcaaa3001?gctgtgcctt?tgagcctata?ctatactgtg?tatgtgtgga?aataaaaatg?tattgtactt3061?ttggagaatt?ttttgtaggc?atttttctgt?cagatttgta?gtaatttgtg?aggtttgtta3121?gagattaata?taggttttct?ttctgtatta?taaaatgcac?caagcaatta?tggtggacct3181?attaccctat?gggtaagaaa?taaatggaaa?tatgacatcg?gatgtttcag?caactgttct3241?gtaaataaaa?tctttgatca?caccactcag?tgtgataatt?gtgtctacag?ctaaaatgga3301?aatagtttta?tctgtacagt?tgtgcaagat?atgaatggtt?tcacactcaa?ataaaaaata3361?ttgaaccccc?aaaaaaaaaa?aa
-10cDNAMNWAAFGGSEFFIPEGIQIDSRCPLSRNITEWYHYYGIAVKESSQYYDVMYSKKGAAWVSETGKKEVSKQTVAYSPRSKLGTLLPEFPNVKDLNLPASLPEEKVSTFIMMYRTHCQRILDTVIRANFDEVQSFLLHFWQGMPPHMLPVLGSSTVVNIVGVCDSILYKAISGVLMPTVLQALPDSLTQVIRKFAKQLDEWLKVALHDLPENLRNIKFELSRRFSQILRRQTSLNHLCQASRTVIHSADITFQMLEDWRNVDLNSITKQTLYTMEDSRDEHRKLITQLYQEFDHLLEEQSPIESYIEWLDTMVDRCVVKVAAKRRGSLKKVAQQFLLMWSCFGTRVIRDMTLHSAPSFGSFHLIHLMFDDYVLYLLESLHCQERANELMRAMKGEGSTAEVREEIILTEAAAPTPSPVPSFSPAKSATSVEVPPPSSPVSNPSPEYTGLSTTGAMQAYTWSLTYTVTTAAGSPAENSQQLPCMRNTHVPSSSVTHRIPVYPHREEHGYTGSYNYGSYGNQHPHPMQSQYPALPHDTAISGPLHYAPYHRSSAQYPFNSPTSRMEPCLMSSTPRLHPTPVTPRWPEVPSANTCYTNPSVHSARYGNSSDMYTPLTTRRNSEYEHMQHFPGFAYINGEASTGWAK
TESTIS SPECIFIC PROTEIN 1's 0 gene open reading frame aminoacid sequence
The objective of the invention is: in view of TESTIS SPECIFIC PROTEIN 1's 0 gene is the brand-new gene that the inventor finds, human at present understanding and the understanding to this gene only limits to the scientific research that the inventor does.Because this gene is the gene relevant with the testis specific function, and be the low expression of embryo, adult's high expression level, therefore this gene is relevant with testicualr development probably, its unconventionality expression may influence the male sex's reproductive function, therefore the research of this gene can be used for: 1. by this gene Fusion albumen of preparation, be used to study the relation of this gene and testicular function, also may become a kind of biologics for the treatment of the male testical relative disease.
2. prepare specific antibody: the specific antibody of gene coded protein is the indispensable important tool of research gene structure and function, the polyclone and the monoclonal antibody of TESTIS SPECIFIC PROTEIN 1's 0 gene coded protein, can be used as immunohistochemical staining, ELISA, immuno-electron microscope, multiple detection method such as co-immunoprecipitation according to the immunology principle design, can be used for location and the quantitative examination of this gene coded protein in histocyte, also can be used for various biological specimens (as blood, seminal fluid, urine etc.) content research in and protein-protein interaction research etc.Therefore, this antibody might be used for preparation and TESTIS SPECIFIC PROTEIN 1's 0 abnormal gene expression relative disease diagnostic kit, and might be used to prepare antifertility drug.
3. prepare gene diagnosis chip: functional gene is used to prepare the importance that gene diagnosis chip is the gene development research, all significant at aspects such as medical diagnosis on disease (as male sterility, sexual disorder) and drug screenings.Technical solution of the present invention
1. the proteic preparation of TESTIS SPECIFIC PROTEIN 1's 0 gene fusion
Nucleotide sequence and plasmid pDEST with TESTIS SPECIFIC PROTEIN 1's 0 gene open reading frame
TM15 are prepared into the GST-SP2-pDEST15 expression plasmid, cultivate 0.5OD600 for 30 ℃ in the LB of no NaCl, and after 0.3M NaCl induced, ultrasonication obtained the fusion rotein of purifying after GST affinity chromatography column purification.
2. TESTIS SPECIFIC PROTEIN 1's 0 gene coded protein specific polyclonal antibody
Getting fusion rotein combines with carrier proteins KLH (Keyhole Limpet Hymocyanin), through the dialysis after again with fully/Freund (Complete/Incomplete Freund ' sAdjuvant) balanced mix, immunity new zealand rabbit (back subcutaneous injection), every 3～4 all booster injections once, totally three times, after three months, get blood system from serum.Be ELISA in contrast with animal serum before the immunity, measure antibody titers in each immunize rabbit serum.Screen high titre serum, further do competition and suppress the specificity that immunity test is determined antibody with antigen.Determine that according to ELISA result its dilution that is used for immunohistochemical staining multiplication number is 1: 200～500.
3. TESTIS SPECIFIC PROTEIN 1's 0 gene coded protein monoclonal antibody specific
Getting fusion rotein combines with carrier proteins KLH (Keyhole Limpet Hymocyanin), after dialysis again with fully/Freund (Complete/Incomplete Freund ' sAdjuvant) mixes, immunity Balb/C mouse (intraperitoneal injection), every other week once, continuous 2-3 time, vein is strengthened once before merging, after ELISA method confirmation production of antibodies, get the spleen separating Morr. cell, merge the generation hybridoma with the myeloma cell then.Positive colony is through subclone repeatedly, until the monoclonal hybridoma strain that obtains anti-TESTIS SPECIFIC PROTEIN 1's 0 antibody of secretion.
4. TESTIS SPECIFIC PROTEIN 1's 0 gene is used to prepare the testicular function gene expression chip
(1) with the nucleotide sequence design primer of TESTIS SPECIFIC PROTEIN 1's 0 gene of obtaining, TESTIS SPECIFIC PROTEIN 1's 0 gene order of amplification total length is as the sample of some film.
(2) with Britain BioRobotics put automatically the film instrument with specimens point on 8 * 12cm nylon membrane, 2 points of each specimens point, several approximately ng of every DNA amount. 8 housekeeping genes of positive control (a.ribosomal protein S9; B.Actin gamma; C.glyceraldehyde-3-phosphate dehydrogenase; D.hypoxanthinephosphoribosyltransferase 1; E.H.sapiens mRNA for 23KD highly basic protein; F.ubiqnitin C; G.Phospholipase A2; H.ubiquitin carboxy I-ferminal esterase L I).Negative control adds lambda bacteriophage dna and P-blue plasmid.
(3) chip that will contain this gene is used for the diagnosis and the drug screening of TESTIS SPECIFIC PROTEIN 1's 0 genetically deficient
33The P mark:
Extracting sample mRNA to be checked or DNA are with Random Primer labelling method mark sample mRNA to be checked or DNA, as the probe of hybridization.
Hybridization:
After the nylon membrane label probe hybridization that point is made, oven dry is shielded compressing tablet with phosphorus in the hybrid heater (68 ℃).
Signal scanning and analysis:
Analyze hybridization signal with registering instrument.According to the strong and weak analyzing influence TESTIS SPECIFIC PROTEIN 10 expression of gene protein level of hybridization signal or the disappearance of TESTIS SPECIFIC PROTEIN 1's 0 gene.
Claims (5)
1. TESTIS SPECIFIC PROTEIN 1's 0 gene (NYD-SP10) is cloned the fusion of preparation; NYD-SP10,：MNWAAFGGSEFFIPEGIQIDSRCPLSRNITEWYHYYGIAVKESSQYYDVMYSKKGAAWVSETGKKEVSKQTVAYSPRSKLGTLLPEFPNVKDLNLPASLPEEKVSTFIMMYRTHCQRILDTVIRANFDEVQSFLLHFWQGMPPHMLPVLGSSTVVNIVGVCDSILYKAISGVLMPTVLQALPDSLTQVIRKFAKQLDEWLKVALHDLPENLRNIKFELSRRFSQILRRQTSLNHLCQASRTVIHSADITFQMLEDWRNVDLNSITKQTLYTMEDSRDEHRKLITQLYQEFDHLLEEQSPIESYIEWLDTMVDRCVVKVAAKRRGSLKKVAQQFLLMWSCFGTRVIRDMTLHSAPSFGSFHLIHLMFDDYVLYLLESLHCQERANELMRAMKGEGSTAEVREEIILTEAAAPTPSPVPSFSPAKSATSVEVPPPSSPVSNPSPEYTGLSTTGAMQAYTWSLTYTVTTAAGSPAENSQQLPCMRNTHVPSSSVTHRIPVYPHREEHGYTGSYNYGSYGNQHPHPMQSQYPALPHDTAISGPLHYAPYHRSSAQYPFNSPTSRMEPCLMSSTPRLHPTPVTPRWPEVPSANTCYTNPSVHSARYGNSSDMYTPLTTRRNSEYEHMQHFPGFAYINGEASTGWAK
2. according to the described polyclonal antibody at the preparation of NYD-SP10 gene open reading frame aminoacid sequence of claim 1, after it is characterized in that fusion rotein and complete and Freund mixing, immune new zealand rabbit prepares polyclonal antibody.
3. according to the described monoclonal antibody of claim 1 at the preparation of NYD-SP10 gene open reading frame aminoacid sequence, it is characterized in that fusion rotein mixes with complete and Freund, immunity Balb/C mouse, the separating spleen bone-marrow-derived lymphocyte and and the myeloma cell merge the generation hybridoma, further, prepare monoclonal antibody with the mono-clonal of subclone method, ELISA and antibody typing method screening IgG secretion antibody.
4. it is characterized in that pcr amplification NYD-SP10 gene cDNA according to claim 1 is described at NYD-SP10 gene open reading frame aminoacid sequence is used to prepare the testicular function gene expression chip, put in chip and be used for chip preparation.
5. according to claim 1, prepare fusion rotein at NYD-SP10 gene open reading frame aminoacid sequence, it is characterized in that fusion rotein can be used as the genomic medicine of treatment relative disease.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01113505 CN1318557A (en) | 2001-04-11 | 2001-04-11 | Human testicular specific protein-10 gene encoded protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01113505 CN1318557A (en) | 2001-04-11 | 2001-04-11 | Human testicular specific protein-10 gene encoded protein |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1318557A true CN1318557A (en) | 2001-10-24 |
Family
ID=4660226
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 01113505 Pending CN1318557A (en) | 2001-04-11 | 2001-04-11 | Human testicular specific protein-10 gene encoded protein |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1318557A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117987376A (en) * | 2024-03-26 | 2024-05-07 | 玛特瑞尔(吉林)生物技术有限公司 | Human sperm acrosin SP-10 monoclonal antibody, hybridoma cell strain and application thereof |
-
2001
- 2001-04-11 CN CN 01113505 patent/CN1318557A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117987376A (en) * | 2024-03-26 | 2024-05-07 | 玛特瑞尔(吉林)生物技术有限公司 | Human sperm acrosin SP-10 monoclonal antibody, hybridoma cell strain and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113355332B (en) | HEG1 gene mutant and application thereof | |
| CN109837345A (en) | Detect the primer and method of mouse cell residual DNA | |
| Scopes | Overview of protein purification and characterization | |
| CN111304226A (en) | Nucleic acid for coding CYP1B1 gene mutant and application thereof | |
| CN1318557A (en) | Human testicular specific protein-10 gene encoded protein | |
| CN1316435A (en) | Human testicular development specificity protein-8 gene coding protein | |
| CN1318559A (en) | Human sperm skelemin gene encoded protein | |
| CN1309138A (en) | Human testicular cytodifferentiation related protein gene coded protein | |
| CN1380338A (en) | Human testis specific protein -16 gene coded protein | |
| CN111662372B (en) | CAPSL mutant gene, reagent, kit and application thereof | |
| CN1318558A (en) | Human testicular development specific protein-12 gene encoded protein | |
| CN1382697A (en) | Human testis development specific protein-28 gene coding protein | |
| CN1319665A (en) | Human testics kinesin 9 gene coding protein | |
| CN1318556A (en) | Human nuclear transcription factor-2 gene encoded protein | |
| CN1319660A (en) | Human testis specific protein kinase gene coding protein | |
| CN1319666A (en) | Human testis development relative gene coding protein | |
| CN1316434A (en) | Human specific protein kinase-15 gene coding protein | |
| CN1318554A (en) | Human testicular development specific protein-11 gene encoded protein | |
| CN1318560A (en) | Human testicular specific protein-1 gene encoded protein | |
| CN1316433A (en) | Human exonuclease factor gene coding protein | |
| CN1382695A (en) | Human testis development specific protein-22 gene coding protein | |
| CN1318555A (en) | Human testicular development relative protein gene encoded protein | |
| CN112345769A (en) | Osteocalcin latex enhanced turbidimetry detection kit based on polyclonal antibody and preparation method thereof | |
| CN1313339A (en) | Human sperm binding gene coded protein | |
| CN1382696A (en) | Human testis development specific protein-25 gene coding protein |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |