人乳头瘤病毒18型E7蛋白基因的克隆研究

Cloning of E 7 Gene of Human Papillomavirus Type 18

根据人乳头瘤病毒ＨＰＶ１６，ＨＰＶ１８的核苷酸序列，设计扩增其Ｅ７基因的特定引物，并使引物的５′端引入ＥｃｏＲＩ，ＨｉｎｄⅢ酶切位点，用聚合酶链反应扩增ＨＰＶ１８Ｅ７片段，通过ｐＵＣ１８质粒载体多聚接头的ＥｃｏＲＩ，ＨｉｎｄⅢ双酶切位点与Ｅ７基因片段连接，构建一新的重组体，转化到大肠杆菌ＪＭ１０９，经筛选，快提质粒等鉴定，初步证实获得了ＨＰＶ１８Ｅ７基因的重组体。

According to the sequence of HPV 16 , HPV 18 DNA, we designed the primers to amplify the E 7 gene and the primers 5’terminal incorporate with restriction endonuclease sites EcoR I and Hind Ⅲ, The ampilfied E 7 gene fragment was inserted into vector plasmid pUC18 at EcoR I/ Hind Ⅲ cloning site of the polylinker .A new recombinant plasmid was constructed and transformated into Escherichia Coli JM 109. The cloning was obtained by screening and other methods of identification,