中国人丙型肝炎病毒核心基因cDNA的克隆与序列分析

Cloning and Sequencing of Core Gene cDNA of Chinese Hepatitis C Vims

利用本室设计的引物对我国人丙型肝炎病毒核心蛋白基因进行了cDNA合成和扩增,获得了一长534bp的cDNA片段,定名为Q534。将此片段克隆到pUC18质粒中,并进行了序列分析。结果表明所获得的cDNA克隆位于Chiron序列的320-853位,此序列与原型相应序列在核苷酸和氨基酸水平上的同源性分别为90.9％和97.6％,与日本HCV-J株和BK株相应序列的核苷酸同源性为96.8％和97.0％;氯基酸的同源性为98.2％和98.8％。序列比较结果表明此序列属HCVⅡ组。

One hepatitis C virus(HCV) cDNA fragment, 534bp in length and designated as Q534, was obtained by means of PCR amplification with self-designed oligonucleotide primers. Q534 was doned into Hinc II site of pUC18 and the- recombinant plasmid pQ534 was then selected from the bacterial transformants. The sequence data indicated the Q534 was a cDNA fragment of HCV core gene, and located in HCV genome from positions 320 to 853 in correspondence with Chiron's prototype sequence. The homologies between Q534 and the prototype at the levels of nucleotides and amino adds were 90.9% and 97.6%, respectively. The homologies of Q534 with Japanese HCV-J and HCV-BK strains were 96.8% and 97.0% at the nucleotide level, and 98.2% and 98.8% at the amino add level. Compared with the corresponding sequences of other HCV isolates, this Chinese HCV isolate we obtained in the present study belongs to HCV group II .