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c D N Acloning and sequence analysis of hepatitis G virus genome isolatedfrom a Chinese blood donor 被引量:1

cDNA cloning and sequence analysis of hepatitis G virus genome isolated from a Chinese blood donor
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摘要 Objective To obtain full length sequence of a Chinese hepatitis G virus (HGV) strain (HGVch) and investigate the genetic characteristic of HGVch and its identity to other isolates Methods Reverse transcription (RT) and nested PCR were used to screen HGV RNA positive serum and amplify cDNA fragments A positive serum without known hepatitis virus markers was selected for isolating HGV RNA template The HGV genome was divided into 12 overlapping fragments and directly cloned into pGEM T vector Sequences were determined by dideoxy terminus end method of DNA sequencing and then analyzed by computer Results The twelve fragments of HGVch cover 9213 nucleotides in length, containing a large open reading frame (ORF) encoding 2873 animo acids polyprotein that began with a methonine residue and ended at termination codon HGVch is about 86 5%-89 5% identical to other known HGV isolates at the nucleotide level and about 93 9%-96 2% at the deduced animo acid level Conclusion HGV is a non A E hepatitis causal agent, proved to be related with posttransfusion hepatitis in all over the world Chinese HGV isolate has very close relationship to other isolates from Africa, Europe, Japan, without significant difference across the entire genome It is suggested that the sequences of HGV isolates are very conservative and the evolution is very slow Objective To obtain full length sequence of a Chinese hepatitis G virus (HGV) strain (HGVch) and investigate the genetic characteristic of HGVch and its identity to other isolates Methods Reverse transcription (RT) and nested PCR were used to screen HGV RNA positive serum and amplify cDNA fragments A positive serum without known hepatitis virus markers was selected for isolating HGV RNA template The HGV genome was divided into 12 overlapping fragments and directly cloned into pGEM T vector Sequences were determined by dideoxy terminus end method of DNA sequencing and then analyzed by computer Results The twelve fragments of HGVch cover 9213 nucleotides in length, containing a large open reading frame (ORF) encoding 2873 animo acids polyprotein that began with a methonine residue and ended at termination codon HGVch is about 86 5%-89 5% identical to other known HGV isolates at the nucleotide level and about 93 9%-96 2% at the deduced animo acid level Conclusion HGV is a non A E hepatitis causal agent, proved to be related with posttransfusion hepatitis in all over the world Chinese HGV isolate has very close relationship to other isolates from Africa, Europe, Japan, without significant difference across the entire genome It is suggested that the sequences of HGV isolates are very conservative and the evolution is very slow
出处 《Chinese Medical Journal》 SCIE CAS CSCD 1999年第8期75-77,共3页 中华医学杂志(英文版)
关键词 HGVch · cloning · sequence analysis HGVch · cloning · sequence analysis
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