痘苗病毒转移载体pSC65-HPV18 E6、E7的构建及鉴定  

Construction and identification of vaccinia virus transfer vector pSC65-HPV18 E6/E7

作　　者：王雪莲[1] 张晓娇[2] 赵峰[3] 安春丽[1] 

机构地区：[1]中国医科大学病原生物学教研室,沈阳110001 [2]盘锦市疾病预防控制中心,盘锦124010 [3]中国医科大学实验动物部,沈阳110001

出　　处：《中国人兽共患病学报》2012年第6期587-591,共5页Chinese Journal of Zoonoses

基　　金：国家自然科学基金资助项目(No.81101989);辽宁省教育厅科研项目(L2010579)~~

摘　　要：目的构建及鉴定含有人乳头瘤病毒(human papillomavirus,HPV)18型E6、E7基因的痘苗病毒转移载体pSC65-HPV18 E6、E7。方法以HPV18型全基因质粒为模板,PCR扩增HPV18 E6、E7基因,克隆到痘苗病毒转移载体pSC65中,获得重组转移载体pSC65-HPV18 E6、E7。重组转移载体转化大肠杆菌,挑取孤立菌落PCR初步筛选。选择阳性菌落提取质粒,酶切及测序鉴定。结果阳性菌落质粒酶切结果显示有340bp、500bp大小的目的基因片段,表明为重组转移载体。测序结果证实重组转移载体包含HPV18 E6、E7基因。结论成功构建包含HPV18 E6、E7基因的痘苗病毒转移载体pSC65-HPV18 E6、E7,研究结果为进一步构建包含HPV18 E6、E7基因的重组痘苗病毒奠定基础。The gene fragments of HPV18 E6 and E7 were amplified by PCR and cloned to vaccinia virus transfer vector pSC6 to construct and identify vaccinia virus transfer vectors pSC65-HPV18 E6 and E75.The recombinant products were transformed into E.coli.The isolated colonies were initially selected by PCR and the positive colonies were selected for the extraction of plasmid.The plasmids were identified by enzyme digestion and gene sequencing.Results of enzyme digestion showed 340bp and 500bp target gene fragments respectively.The recombinant vectors containing HPV18 E6 and E7 genes were demonstrated by gene sequencing.It＇s concluded that the recombinant vaccinia virus transfer vectors pSC65-HPV18 E6 and E7 were constructed successfully and this study lays foundation for the future construction of recombinant virus.

关 键 词：人乳头瘤病毒 痘苗病毒 转移载体pSC65 

分 类 号：R373[医药卫生—病原生物学]