【摘要】 利用逆转录套式ＰＣＲ扩增Ⅲ型中国株ＨＣＶＥ２／ＮＳ１基因片段，将其克隆到ｐｃＤＮＡ３载体上．采用双脱氧链终止法测定插入片段的核苷酸序列．并与已知分离株的相应区域进行同源性比较．首次克隆出Ⅲ型中国株ＨＣＶＥ２／ＮＳ１基因（ＨＣ－Ｗ１４），其核苷酸序列与Ⅲ型日本株ＨＣＶ（ＨＣ－Ｊ６）该区域同源性为８８．３７％，其推定的氨基酸同源性为８９．２９％．而与已知的非Ⅲ型株ＨＣＶ该区域相比，核苷酸及氨基酸的同源性均相对较低．Ⅲ型中国株ＨＣＶ与Ⅱ型中国株ＨＣＶ在Ｅ２／ＮＳ１区域有较大的变异，揭示研制我国的ＨＣＶ疫苗应该考虑这种基因型之间的变异性．更多还原

【Abstract】 In order to clone and sequence E2/NS1 gene of hepatitis C virus(HCV),E2/NS1 gene derived from a genotype Ⅲ Chinese isolate of HCV was amplified by reverse transcription polymerase chain reaction and inserted into vector pcDNA3.Dideoxy chain termination methods were employed to sequence E2/NS1 gene and homologies to corresponding region of reported isolates were analyzed.E2/NS1 gene derived from a genotype Ⅲ Chinese isolate of HCV was cloned for the first time and named HC W14.HC W14 was nearly full length E2/NS1 gene of HCV encoding 364 amino acids (aa378 aa741,not including initiator and terminator codon).Identity of HC W14 in nucleotide and putative amino acid to that of a genotype Ⅲ Japanese isolate of HCV(HC J6) was 88 37% and 89 29%.Homologies to that of non type Ⅲ isolates were relatively low.Especially,homologies in hypervariable region of E2/NS1 were only 44.44%—59.26%.It was concluded that high variation existed in E2/NS1 region between genotype Ⅲ and Ⅱ Chinese isolates of hepatitis C virus.The variability of E2/NS1 gene should be taken into account in the development of vaccine against HCV in China.更多还原