【摘要】 目的　了解丙型肝炎病毒 (HCV) 2a型基因组E1E2 NS1区序列 ,为进一步研究HCV包膜蛋白的生物学功能奠定基础。方法　用逆转录套式PCR(RT nPCR)从江苏省 1例丙型肝炎患者血清中扩增出两条分别约 770bp、110 0bp的片段 ,分别以限制性内切酶EcoRⅠ、BamHⅠ和EcoRⅠ、HindⅢ双酶切后连入pUC19载体中 ,转化感受态细胞 ,经限制性酶切长度多态性分析 (RFLP)和PCR法证实为阳性克隆后 ,用全自动序列分析仪测序。结果　分别测得约 770bp、110 0bp的核苷酸序列 ,拼接后得到的完整核苷酸序列及其编码的氨基酸序列分别与HCV 1、HC C2、HCV BK、HC J6、HC J8相应序列作比较 ,显示分离株HCV在核苷酸水平上与以上分离株的同源性分别为 60 .5 %、60 .1%、5 9 7%、87 8%、67.5 % ;在氨基酸水平上的同源性分别为 67.3 %、66.4%、65 .0 %、87 8%、79.0 %。结论　分离株 (HCV JS)与HC -J6同属 2a型 ,但同源性有一定差异更多还原

【Abstract】 Objective To investigate sequence of the complete E region of genotype 2a hepatitis C virus genome and to set up the basis for the further study on biological functions of envelope proteins of HCV. Methods Two cDNA fragments of 770bp,1 100bp long,from serum of a patients infected HCV, were amplified by reverse transcription nested polymerase chain reaction (RT nPCR). After being digested with two restriction endonucleases,the two fragments were cloned into JM105 respectively, then sequenced. Results The two corresponding nucleotide sequences were obtained and ligated. The complete nucleotide sequences of envelope region and corresponding deduced amino acid sequences were compared with those of HCV 1,HC C2,HCV BK,HC J6 and HC J8. The homology of nucleotide sequence of the isolated strains was 60.5%, 60.1%, 59.7%, 87.8% and 67.5% respectively; the amino acid sequence homology to the isolated strains was found to be 67 3%,66.4%,65.0%,87.8%,79.0%, respectively. Conclusion The results indicate that the isolated strain (HCV JS) belongs to genotype 2a, but there is heterogeneity between HCV JS and HC J6 strain.更多还原