CAJ | 학술논문

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.
목적 극륭화표체절단적병형간염병독(hepatitis C virus,HCV)핵심구기인,위제비HCV핵심구항체준비항원.방법 근거연건분석선취대유우세항원표위적HCV핵심구다태,조출대응DNA서렬,역전록PCR확증목적기인,병장기극륭도원핵표체재체중진행유도표체.표체적목적단백순화후용간접ELISA법검측면역활성.결과 획득료서렬정학적HCV목적기인편단.표체적목적단백위가용성단백,능피흔호지순화.사용해순화단백검측각충양본,결과HCV음성양본적평균흡광도(A)치위0.081,HCV양성양본여음성대조적A치지비균＞2.1,을형간염화매독양성양본적A치도재0.101이하.결론 성공지극륭화표체료HCV핵심구소편단기인,획득료구유량호항원성적절단HCV핵심구다태.
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.