人乳头瘤病毒16L1原核表达质粒的构建  

Constrction of prokaryotic plasmid of HPV16 L1

作　　者：左凤琼[1] 赵素兰[2] 李婉宜[1] 李晓红[3] 吕梅励[1] 蒋中华[1] 李明远[1] 欧阳运薇[4] 

机构地区：[1]四川大学华西基础医学与法医学院 [2]四川省肿瘤医院 [3]四川大学华西药学院 [4]四川大学华西附二院,四川成都610044

出　　处：《四川生理科学杂志》2008年第4期152-153,共2页Sichuan Journal of Physiological Sciences

基　　金：四川省科技厅基金2006J13-100资助

摘　　要：目的:构建人乳头瘤病毒16L1的原核表达质粒,为下一步探讨蛋白的表达以及蛋白的功能研究作准备。方法:以临床HPV16阳性标本为模板,PCR扩增L1的片段,L1片段经EcoRⅠ和SalⅠ双酶切后,插入载体pGEX4T-1,转化JM109感受态细胞,平板筛选获得pGEX4T-1/HPV L1的阳性质粒。通过酶切、测序验证质粒的正确性。结果:构建了原核表达质粒pGEX4T-1/HPV16L1,并通过酶切、测序等方法验证其完全正确。结论:成功构建的pGEX4T-1/HPVL1为今后的蛋白的表达和功能研究打好了坚实的基础,为HPV16的疫苗研究提供了有益的支持。Objective. To construct prokaryotic plasmid containing human papillomavirus（HPV）16 L1 gene and to be basis for expression of HPV16 L1 protein. Methods.. HPV16 L1 gene was amplified by PCR from clinical isolated HPV 16 and inserted into the plasmid pGEX4T-1 followed by EcoR Ⅰ and Sal l. The recombinant plasmld pGEX4T-1/HPV16L1 was transformed into E. coli JM109 and selected with ampicillin. The positive clones were virified by restriction endonucleases EcoR Ⅰ and Sal Ⅰ and sequence analysis. Results and Conclusion. The prokaryotic plasmid pGEX4T-1/HPV16L1 was constructed successfully and may facilitate subse- quent research of the expression and function of HPV16 L1 protein.

关 键 词：人乳头瘤病毒16 质粒 构建 

分 类 号：R346[医药卫生—基础医学] R737.33