我国非肠道传播非甲非乙型肝炎(丙型)病毒NS3蛋白的部份cDNA克隆及序列分析

CLONING AND SEQUENCING OF PARTIAL cDNA OF NS3 PROTEIN OF HCV DERIVED FROM SERA OF CHINESE PATIENTS

用15ml非肠道传播非甲非乙型肝炎病人的混合血清提取病毒RNA,经逆转录和聚合酶链反应(PCR)扩增,获得583个核苷酸的非肠道传播非甲非乙型肝炎病毒(HCV)NS3蛋白的cDNA片段.该片段与美国报道的同片段HCV cDNA原型比较,核酸序列同源性为80.9％,氨基酸序列同源性为93.2％。与日本报道的同片段J1 HCV cDNA相比较,核酸序列和氨基酸序列的同源性分别为92.6％和95.2％。用α-^(32)P同位素标记该片段,与HCV病人血清出现杂交反应。

Hepatitis C virus ( HCV ) RNA was extracted from 15ml mixed sera of HCY patients in China. The cDNA fragment of 533 nucleotides was synthesized and subsequently amplified by the PCR method using specific oligo-nucleotide primers. The sequence of cDNA showed 80.9% homology at the nucleotide level and 93.2% homology at the amino acid level when compared with that of the prototype HCV PT cDNA. It showed 92.6% homology at the nucleotide level and 95.2% homology at the amino acid level when compared with that of HCV J1 cDNA in Japan. Dot blot hybridization of HCV RNA isolated from HCV patient samples with the resulting cDNA fragment labeling by 22P-dCTP was positive in 3 out of 4 cases.