用连续长片段RT-PCR扩增HCV结构基因

Amplification of HCV structure genes by continuous long RT-PCR

目的与方法 :使用连续长片段 RT- PCR,扩增 HCV的结构基因。 结果 :利用连续长片段 RT- PCR技术 ,结合热启动和两段 PCR一次扩增出长 2 .3kb的 HCV结构基因和部分调控序列 ,并用测量分子质量和限制性酶切的方法进行了鉴定。 结论 :利用长片段 RT- PCR可一次扩增出 C、E1、E2基因和部分 5 - N TR。

Objectives and Methods: The structure genes of HCV was amplified with continuous long RT PCR. Results:A 2.3kb DNA fragment was amplified from HCV RNA by continuous long RT PCR technology combined with Hot Start and shuttle PCR methods in only one step. By restriction enzyme digestion and determination of molecular weight, the product was confirmed to be the needed DNA sequence, which are HCV structure genes and part of regulating sequence. Conclusions:By continuous long RT PCR,C,E 1,E 2 genes and part of 5 NTR of HCV can be amplified in only one experiment.