HCVE2区基因的分子克隆及序列分析

Molecular Cloning and Sequencing of HCV E2 Gene

用RT PCRKIT从西安血站大样抗HCV阳性血清中筛选出HCVRNA阳性血清 ,提取HCV的RNA ,利用随机引物反转录合成其cDNA并进行半巢式PCR反应。将纯化的PCR产物酶切后与表达载体PET 2 2b+连接 ,经过双脱氧末端终止法双向测序 ,得到 85 2bp长的核苷酸序列。通过将该序列与已知不同型的HCVE2序列比较得知 。

HCV RNA positive serum was first selected by RT PCR test kit from several anti HCV positive sera obtained from Xi'an.HCV RNA extracted from the elected sera was converted to cDNA by reverse transcription with random primer.Half nested PCR was performed.The amplified product was 852 bp.The purified PCR product was digested by restriction endonucleases and then ligated to epressio vector pET 22b\++.Its nucleotide sequence was determined by dideoxy chain termination method.A comparison of the sequence with several isolates reported previously showed that the sequence belonged to HCV type Ⅱ.