摘要
应用聚合酶链式反应技术(PCR)扩增轮状病毒VP7基因,并将其克隆到pMD18-T simple载体上,对重组子进行PCR检测和限制性内切酶分析,并测定DNA全序列。结果显示,克隆片段全长为981 bp。将轮状病毒VP7基因定向的克隆到原核表达载体pET-32a启动子下游,构建原核表达载体pET-32aVP7。将质粒pET-32aVP7转化Transetta表达菌株进行诱导表达,裂解菌体细胞抽提蛋白质进行SDS-PAGE。结果表明,轮状病毒壳蛋白VP7基因在Transetta表达菌株内得到成功表达。
Rotavirus gene VP7 was obtained by PCR techinque and cloned into pMD18-T simple.The recombinant clone was detected by PCR technique and analysed by the restriction enzyme.A full length of cDNA gene was sequenced.The results showed that the length of the clone sequence was 981 bp.Rotavirus gene VP7 was cloned into promoter downstream of expression vector pET-32a in orientation.A expression vector pET-32aVP7 was constructed.The expression of gene VP7 was induced by 1 mmol/L IPTG in Transetta cells.The lysate of E.coli cells was analyzed by electrophoresis and staining of polyacrylamide gel by coomassie brilliant blue R-250 showed a VP7 band of molecular weight 57 kD.
出处
《生物技术通报》
CAS
CSCD
北大核心
2011年第3期160-162,174,共4页
Biotechnology Bulletin
基金
辽宁省教育厅项目(2008T023)