摘要
目的 了解中国大陆丙型肝炎病毒 (HCV) 2a型基因组 5’非编码区的核苷酸序列 ,研究其序列变异特征。方法 以限制性片段长度多态性分析法 (RFLP)初步筛选出 2例 2a型HCV感染者 ,用逆转录套式PCR(RT nPCR)法从其血清中分别扩增出长约 60 0bp的片段 ,以限制性内切酶EcoRI和BamHI双酶切后克隆于 pUC1 9载体中 ,转化感受态细胞 ,以RFLP法鉴定为阳性克隆后 ,用全自动序列分析仪测序。结果 2条分离株 (HCV JFB1 ,HCV JFB2 )均测得约 60 0bp的核苷酸序列 ,将 5’端 2 80个核苷酸分别与HCV 1、HC C2、HC J5、HC J6、HC J85’UTR相应序列作比较 ,其同源性在 92 8%～ 99 6 %。同时发现分离株也存在多个碱基突变。结论 HCV5’UTR高度保守 ,2a型HCV基因突变以碱基的转换为主 ,其中一分离株 (HCV JFB1 )
Objective To investigate nucleotide sequence of 5 UTR of genotype 2a hepatitis C virus genome and study the variation of the sequence.Methods Two patients infected with genotype 2a HCV were chosen by restriction fragment length polymorphism.From their sera,the cDNA fragment of about 600bp in length was amplified by reverse transcription nested polymerase chain reaction (RT nPCR).After digested with two restriction endonucleases,the two fragments were transformed into JM105,respectively,then sequenced.Results 280 bases of 5 UTR of two nucleotide sequences were obtained and compared with those of HCV 1,HC C2,HC J5,HC J6 and HC J8.The homologies of 5 UTR of the isolated strains are higher than 92 8%.Several base variations were found in these HCV strains.Conclusion Nucleotide sequence of HCV5 UTR is highly conserved and base conversions are vital variations in 5 UTR of these HCV strains.A base deletion was found in HCV JFB1.
出处
《江苏医药》
CAS
CSCD
北大核心
2002年第12期886-888,共3页
Jiangsu Medical Journal
基金
国家自然科学基金 ( 39770 6 84 30 170 84 4)
江苏省青年科技基金 (BQ 2 0 0 0 0 2 3)
