融合基因Tat-HPV16L1的克隆及其在酿酒酵母中的表达  

Cloning and Expression of Fusion Gene Tat-HPV16L1 in Saccharomyces cerevisia

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

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

出　　处：《热带医学杂志》2009年第6期597-600,共4页Journal of Tropical Medicine

基　　金：广东省重点实验室基金(No.960327)

摘　　要：目的构建人乳头瘤病毒16型L1基因和蛋白转导肽的Tat基因的融合表达载体,导入酿酒酵母表达系统进行诱导表达,为口服基因疫苗的制备打下基础。方法以pBR322-HPV16质粒为模板扩增L1基因,与酿酒酵母表达载体pYES2连接,重组质粒再与自行设计的蛋白转导功能片段Tat连接,经测序鉴定后转入酿酒酵母表达菌株INVSc1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,经过半乳糖诱导表达后进行SDS-PAGE电泳和Western blotting分析,结果表明诱导表达成功。结论成功构建和表达了融合表达载体pYES2-Tat-HPV16L1,为制备以酿酒酵母为运送载体的口服人乳头瘤病毒疫苗打下基础。Objective To clone the recombinant fusion gene Tat-HPV16L1 and to express the gene in Saccharomyces cerevisia. Method The L1 gene of HPV16 was amplified from pBR322-HPV16 plasmid by PCR. Tat gene was synthesized and the recombinant plasmid pYES2-Tat-HPV16L1 was established and sequenced. Fusion gene Tat-HPV16L1 was induced to express in host cell INVSc1 by galactose. The expressed recombinant protein was identified by SDS-PAGE and Western-blot. Result The Saccharomyces cerevisia expression system pYES2-Tat- HPV16L1 was established and the fusion gene Tat-HPV16L1 was successfully expressed. Conclusion The yeast based plasmid carrying the cloned Tat-HPV16 fragment may be used for the future development of oral vaccine for the prevention of human papillomavirus infection.

关 键 词：人乳头瘤病毒 L1基因 TAT 酿酒酵母 口服基因疫苗 

分 类 号：R737.33[医药卫生—肿瘤]