人乳头瘤病毒16型转化基因的筛选及次级克隆  

SELECTION AND SUBCLONE OF THE TRANSFORMING GENES OF HUMAN PAPILLOMAVIRUS TYPE 16

作　　者：袁育康[1] 董小平[1] 房益兰[1] 刘延娜[1] 楚雍烈[1] 任会勋[1] 

机构地区：[1]西安医科大学微生物学教研室,西安710061

出　　处：《西安医科大学学报》1992年第2期119-122,共4页Journal of Xi'an Medical University(Chinese)

基　　金：国家自然科学基金

摘　　要：人乳头瘤病毒16型转化作用在宫颈癌发生中可能起重要作用,其转化基因主要位于基因组早期区域E6、E7开放阅读框架内。为了进一步研究HPV16转化作用,我们以Pstl+ECoRI酶解HPV16DNA,以pUC-19为载体,大肠杆菌JM103细胞为宿主菌,经两次定向次级克隆,组建了质粒pEP-8。经限制性核酸內切酶和Southem转印杂交证实,其插入片段为-1.43Kb带有E6、E7 ORF的DNA片段。在E6区上游序列还含有两个TATA盒,1个Cat盒和1个细胞特异性增强子。该质粒的组建为检测HPV感染细胞中mRNA转录提供了特异性的早期基因探针,同时为进一步研究HPV转化作用及转化蛋白打下了基础。The transformation of human papillomavirus type 16 (HPV16) may play somc important roles in the oncogenesis of cervical carcinoma Studies showed that the transforming gence mainly located in the E6. E7 open reading frames (ORFs). We digested HPV16 DNA with PstI+EcoRI. Using pUC-19 as the vector, E. coli JM103 strain as the host cell, after two times of directive subelone, plasmid pEP-8 was construeted, confirmed by RE analysis and Southern blot hybridization, the inserted band of pEP-8 was 1.43Kb long contained the intact E6 E7 ORFs, and also a cat-box region, two TATA-boxics and a cell-type specific cnhancer in the upstream swquence. The pEP-8 could be used as the specific probe of HPV-16 E region for identifing virol mRNAs in the HPV infected cells. Meanwhile it was a basework for studying the transformation and transforming proteins of HPV-16.

关 键 词：人乳头瘤病毒 次级克隆 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤]