[Cloning and sequencing of E2/NS1 gene from a Chinese genotype III isolate of hepatitis C virus].
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abstract
OBJECTIVE: To clone E2/NS1 gene from a Chinese genotype III isolate of hepatitis C virus into mammalian expression vector and analyze its primary nucleotide structure. METHODS: Reverse transcription nested-PCR methods were employed for amplification E2/NS1 gene which was cloned into mammalian expression vector pcDNA3 by directed clone. Sequence of inserted gene was analyzed by Sanger's method. RESULTS: E2/NS1 gene from a Chinese genotype III isolate of hepatitis C virus was cloned into eukaroytic expression vector effectively. Sequence of E2/NS1 showed 88.37% identity in nucleotide and 89.29% in putative amino acid to that of a Japanese genotype III isolate of hepatitis C virus. Nevertheless it showed 70.69% and 75.55% identity to that of a Chinese genotype II isolate of hepatitis C virus. CONCLUSION: E2/NS1 gene from a Chinese genotype III isolate of hepatitis C virus was successfully cloned into mammalian expression vector by technique of DNA recombination. Sequence analysis showed that E2/NS1 gene has good homology intra-genotype and poor homology intergenotype of hepatitis C virus. Research of primary nucleotide structure of different genotype hepatitis C virus might be helpful for the development of vaccine against the virus.
