截短的丙型肝炎病毒核心区基因的克隆和表达

Cloning and expression of truncated gene of hepatitis C virus core region

目的 克隆和表达截短的丙型肝炎病毒（hepatitis C virus,HCV）核心区基因,为制备HCV核心区抗体准备抗原.方法 根据软件分析选取带有优势抗原表位的HCV核心区多肽,找出对应DNA序列,逆转录PCR扩增目的基因,并将其克隆到原核表达载体中进行诱导表达.表达的目的蛋白纯化后用间接ELISA法检测免疫活性.结果 获得了序列正确的HCV目的基因片段.表达的目的蛋白为可溶性蛋白,能被很好地纯化.使用该纯化蛋白检测各种样本,结果HCV阴性样本的平均吸光度（A）值为0.081,HCV阳性样本与阴性对照的A值之比均＞2.1,乙型肝炎和梅毒阳性样本的A值都在0.101以下.结论 成功地克隆和表达了HCV核心区小片段基因,获得了具有良好抗原性的截短HCV核心区多肽.

Objective To clone and express truncated hepatitis C virus （HCV） core gene for preparing antigens used for production of HCV-core antibody. Methods HCV-core polypeptide with predominant antigenic determinants was selected by software analysis. The corresponding DNA sequence was amplified by reverse transcription PCR. Then, the gene was cloned into a prokaryotic expression vector for expressing HCV-core antigen. The expressed antigen was purified and detected by indirect ELISA. Results A target gene with correct DNA sequence was obtained. The expressed antigen was a soluble protein and purified for detection of different samples. The results showed that the average absorbance （A） value of HCV-negative samples was 0. 081, ratio of A values for HCV-positive samples to negative samples were 〉 2.1, and A values of hepatitis B-and syphilis-positive samples were 〈 0. 101. Conclusions HCV-core gene is successfully cloned. The expressed HCV core polypeptide has good antigenicity.