人乳头瘤病毒11型早期蛋白E6和E7基因的克隆及序列分析  

Molecular cloning and sequence analysis of E6 and E7 gene of human papillomavirus type 11

作　　者：王飞[1] 王群[2] 毕志刚[1] 李光富[3] 王新军[3] 张兆松[3] 

机构地区：[1]南京医科大学第一附属医院皮肤科,南京210029 [2]广东省人民医院皮肤科,广州510080 [3]南京医科大学分子生物研究所,南京210029

出　　处：《中国麻风皮肤病杂志》2005年第2期81-84,共4页China Journal of Leprosy and Skin Diseases

基　　金：江苏省重点学科基金(135-03)

摘　　要：目的:从尖锐湿疣(CA)标本中克隆出人乳头瘤病毒11型(HPV11)早期蛋白E6、E7基因,并进行序列测定及编码氨基酸序列分析,为HPV感染的检测和基因工程疫苗研究奠定基础。方法:用PCR法,从CA标本中扩增出HPV11E6、E7基因,与载体pGEX-6P-1连接成重组质粒pGEX-6P-1E6、pGEX-6P-1E7,酶切鉴定及双脱氧法测序观察其变异状况。结果:克隆出HPV11早期蛋白E6、E7基因,成功构建重组质粒pGEX-6P-1E6、pGEX-6P-1E7。本研究克隆出的HPV11E7基因与GenBank标准株序列完全相同,E6基因有两个位点的变化。结论:本研究克隆出的HPV11E6、E7基因与标准株基本相同,这将为进一步研究E6、E7基因的表达、免疫活性及流行病学奠定基础。Objective:To clone and analyze the early gene E6, E7 of human papillomavirus type 11(HPV11) to provide the basic information on the detection of HPV infection, and to develop a prophylactic and therapeutic vaccine against HPV. Methods: HPV DNA was extracted from samples of condyloma acuminatum (CA) and HPV11 E6, E7 genes were amplified by PCR. HPV11 E6, E7 genes were cloned into vector pGEX-6P-1 to form pGEX-6P-1/E6, pGEX-6P-1/E7 plasmid, and then, sequenced. Results: The recombinant plasmids pGEX-6P-1/E6 and pGEX-6P-1/E7 were correctly constructed and confirmed with enzyme digestion and sequencing. The nucleotide homology analysis indicated that the E7 gene was equal to the prototype accepted by GenBank and there was 2-point mutation in HPV11 E6 gene. Conclusion: The HPV11 E6, E7 genes have been cloned, which is the foundation of the further study of the immunological activity and epidemiology.

关 键 词：E7基因 HPV11 人乳头瘤病毒11型 早期 克隆 蛋白 重组质粒 标本 E6基因 HPV感染 

分 类 号：R373[医药卫生—病原生物学] R752.53[医药卫生—基础医学]