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Article: cDNA cloning of c33-c antigen gene derived from NS3 region of Chinese HCV genome, expression in Escherichia coli and development of HCV EIA second-generation diagnostic kit.

TitlecDNA cloning of c33-c antigen gene derived from NS3 region of Chinese HCV genome, expression in Escherichia coli and development of HCV EIA second-generation diagnostic kit.
Authors
Issue Date1994
Citation
Science In China. Series B, Chemistry, Life Sciences & Earth Sciences, 1994, v. 37 n. 2, p. 190-202 How to Cite?
AbstractA cDNA fragment of about 860 bp corresponding to the c33-c gene in the non-structural region 3 (NS3) of HCV genome was obtained from one plasma derived from a Chinese HCV carrier who came from Tai' an of Shandong Province, China by the application of reverse transcription (RT) and polymerase chain reaction (PCR) techniques. After the sequence of the cDNA fragment was determined and compared with the equivalent region of the HCV-I (HCV-US) and HCV-II (HCV-BK) genomes, the nucleotide/amino acid sequence homologies were found to be 79.2%/91.3%/ and 91.3%/93.9%, respectively. The prokaryotic expression vector pBV220 was employed for the overproduction of c33-c native recombinant protein in E. coli cells. The expression products were detected by enzyme-linked immunosorbent assay (ELISA) and Western blotting with antisera of chronic hepatitis C patients, and a molecular weight 31 kD of c33-c viral protein was shown to account for 14% of the total cellular soluble proteins. This product was extracted from the bacterial lysate by lysozyme, Triton X-100 and urea treatment, and purified through ion exchange chromatography. The purified c33-c protein combined with a branch peptide MAP-C-19 representing immunodominant epitopes on the nucleocapsid region of HCV genome was used to develop a Chinese HCV EIA 2nd-generation diagnostic kit for the detection of anti-HCV antibodies. Its specificity, sensitivity and reproducibility were all in keeping with the indexes of the national standard for the quality control of the HCV diagnostic kit. The agreement rate between our kit and Abbott company's HCV EIA second-generation diagnostic kit was 99.33%, and the identified rate of positive anti-HCV of our kit was 2% more than that of the Abbott company's kit.
Persistent Identifierhttp://hdl.handle.net/10722/147389
ISSN
1998 Impact Factor: 0.479
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorYang, YPen_US
dc.contributor.authorLiu, CBen_US
dc.contributor.authorJin, DYen_US
dc.contributor.authorZhan, MYen_US
dc.contributor.authorTang, Qen_US
dc.contributor.authorXia, NSen_US
dc.contributor.authorCao, JYen_US
dc.contributor.authorLi, JYen_US
dc.date.accessioned2012-05-29T06:03:22Z-
dc.date.available2012-05-29T06:03:22Z-
dc.date.issued1994en_US
dc.identifier.citationScience In China. Series B, Chemistry, Life Sciences & Earth Sciences, 1994, v. 37 n. 2, p. 190-202en_US
dc.identifier.issn1001-652Xen_US
dc.identifier.urihttp://hdl.handle.net/10722/147389-
dc.description.abstractA cDNA fragment of about 860 bp corresponding to the c33-c gene in the non-structural region 3 (NS3) of HCV genome was obtained from one plasma derived from a Chinese HCV carrier who came from Tai' an of Shandong Province, China by the application of reverse transcription (RT) and polymerase chain reaction (PCR) techniques. After the sequence of the cDNA fragment was determined and compared with the equivalent region of the HCV-I (HCV-US) and HCV-II (HCV-BK) genomes, the nucleotide/amino acid sequence homologies were found to be 79.2%/91.3%/ and 91.3%/93.9%, respectively. The prokaryotic expression vector pBV220 was employed for the overproduction of c33-c native recombinant protein in E. coli cells. The expression products were detected by enzyme-linked immunosorbent assay (ELISA) and Western blotting with antisera of chronic hepatitis C patients, and a molecular weight 31 kD of c33-c viral protein was shown to account for 14% of the total cellular soluble proteins. This product was extracted from the bacterial lysate by lysozyme, Triton X-100 and urea treatment, and purified through ion exchange chromatography. The purified c33-c protein combined with a branch peptide MAP-C-19 representing immunodominant epitopes on the nucleocapsid region of HCV genome was used to develop a Chinese HCV EIA 2nd-generation diagnostic kit for the detection of anti-HCV antibodies. Its specificity, sensitivity and reproducibility were all in keeping with the indexes of the national standard for the quality control of the HCV diagnostic kit. The agreement rate between our kit and Abbott company's HCV EIA second-generation diagnostic kit was 99.33%, and the identified rate of positive anti-HCV of our kit was 2% more than that of the Abbott company's kit.en_US
dc.languageengen_US
dc.relation.ispartofScience in China. Series B, Chemistry, life sciences & earth sciencesen_US
dc.subject.meshAmino Acid Sequenceen_US
dc.subject.meshAntigens, Viral - Geneticsen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCloning, Molecularen_US
dc.subject.meshDna, Complementary - Geneticsen_US
dc.subject.meshDna, Viral - Geneticsen_US
dc.subject.meshEscherichia Coli - Genetics - Metabolismen_US
dc.subject.meshGene Expressionen_US
dc.subject.meshGenome, Viralen_US
dc.subject.meshHepacivirus - Geneticsen_US
dc.subject.meshHepatitis C - Diagnosisen_US
dc.subject.meshHepatitis C Antigensen_US
dc.subject.meshHumansen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshPeptide Fragments - Geneticsen_US
dc.subject.meshReagent Kits, Diagnosticen_US
dc.subject.meshRecombinant Proteins - Biosynthesisen_US
dc.subject.meshSequence Homologyen_US
dc.titlecDNA cloning of c33-c antigen gene derived from NS3 region of Chinese HCV genome, expression in Escherichia coli and development of HCV EIA second-generation diagnostic kit.en_US
dc.typeArticleen_US
dc.identifier.emailJin, DY:dyjin@hkucc.hku.hken_US
dc.identifier.authorityJin, DY=rp00452en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid7520701-
dc.identifier.scopuseid_2-s2.0-0028376884en_US
dc.identifier.scopuseid_2-s2.0-43949151576-
dc.identifier.volume37en_US
dc.identifier.issue2en_US
dc.identifier.spage190en_US
dc.identifier.epage202en_US
dc.identifier.isiWOS:A1994MZ80000007-
dc.identifier.scopusauthoridYang, YP=7409384001en_US
dc.identifier.scopusauthoridLiu, CB=7409795920en_US
dc.identifier.scopusauthoridJin, DY=7201973614en_US
dc.identifier.scopusauthoridZhan, MY=7101690249en_US
dc.identifier.scopusauthoridTang, Q=7201632141en_US
dc.identifier.scopusauthoridXia, NS=35187953700en_US
dc.identifier.scopusauthoridCao, JY=14036761700en_US
dc.identifier.scopusauthoridLi, JY=36065525800en_US

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