从尖锐湿疣中克隆人乳头瘤病毒基因组的初步研究

Primary Study of Molecular Cloning of Human Papillomavirus Genome from Condyloma Acuminata

为了探讨人乳头瘤病毒在重庆地区尖锐湿疣组织中的存在情况,本实验将数个尖锐湿疣标本DNA混合抽提7～9kb DNA片段,并使之与pUC19载体相连。然后利用快速细胞破碎法及α-互补原理筛选重组子,以HPV6b DNA为探针进行斑点杂交,获得pHPV-CA阳性克隆。以HPV-CADNA为探针与尖锐湿疣DNA进行Southern印渍杂交,结果显示HPV-CA在重庆地区尖锐湿疣组织中的存在具有普遍性,并发现HPV-CA以游离形式存在于宿主细胞中。

To investigate the prevalence of human papillomavirus (HPV) in condyloma acuminata in Chongqing,the 7 - 9kb DNA fragment was extracted from several mixed DNA samples of condyloma acuminata and ligated with pUC19 vector. Recombinant plasmids were selected by a-completation and Cracking gel methods. Using HPV6b DNA as probe,Dot blot hybridization was carried out to identify the homologous DNA. A positive clone named pHPV-CA was obtained. It was found that HPV-CA DNA hybridized with all the 14 tested condyloma acuminata DNA suggesting high prevalence of HPV-CA in Chongqing,and HPV-CA in host cell was existed as an episomal form.