融合基因Tat-HPV16L1的克隆及在大肠杆菌中的表达  

Construction of fusion gene Tat-HPV16L1 and its expression in Escherichia coli

作　　者：张进芳[1] 马文丽[1] 危敏[1] 赵慧[1] 朱秀兰[1] 郑文岭[2] 

机构地区：[1]南方医科大学基因工程研究所,广州市510515 [2]华南基因组研究中心,广州市510800

出　　处：《实用医学杂志》2009年第10期1539-1541,共3页The Journal of Practical Medicine

基　　金：广东省重点实验室基金资助项目(编号:960327)

摘　　要：目的:构建人乳头瘤病毒16型L1基因和蛋白转导肽基因Tat的融合表达载体,以大肠杆菌作为表达系统,探索制备经济而高效的人乳头瘤病毒疫苗的新方法。方法:以pBR322-HPV16质粒为模板扩增L1基因,与原核表达载体pET28a连接,重组质粒再与自行设计蛋白转导功能片段Tat连接,经测序鉴定后转入大肠杆菌表达菌株DE3进行诱导表达Tat-HPV16L1融合蛋白。结果:融合基因Tat-HPV16L1测序结果与预期完全符合,经过IPTG诱导表达后进行SDS-PAGE电泳分析,结果诱导表达成功。结论:本研究为制备经济而高效的人乳头瘤病毒疫苗打下了基础。Objective To construct an expression vector containing fusion gene Tat-HPV16L1 and explore a new approach for preparation of an inexpensive yet effective vaccine for human papillomavirus. Methods HPV-16 L1 gene was amplified from the pBR322-HPV16 plasmid by PCR and bound to the prokaryotic expression vector pET28a. The recombinant plasmid pET28a-HPV16L1 was then bound with self-developed gene Tat. After sequencing, the new plasmid was transfected E.coli DE3 expression strain to induce the expression of fusion protein Tat- HPV16L1. Results The finding of fusion gene Tat-HPV16LI sequencing was identical to the expectation outcome.SDS-PAGE showed that Tat-HPV16L1 was expressed successfully after induction with IPTG. Conclusion The successful construction of fusion gene Tat-HPV16L1 lays the foundation for preparing an inexpensive yet highly effective human papillomavirus vaccine.

关 键 词：乳头状瘤病毒属 L1基因 TAT 融合基因 

分 类 号：R346[医药卫生—基础医学]