腺病毒载体基因转移系统的建立及其在脑肿瘤中的高效表达

构建了含有CMV启动子调控的HSV－tk基因的重组质粒pAdCMVtk，将其与缺陷型腺病毒Ad5大框架pJM17一起共转染病毒包装细胞293细胞，获得有感染能力但复制缺陷的重组腺病毒AdCMVtk，以其感染大鼠C5脑胶质瘤细胞后用PCR方法证实HSV-tk基因能被腺病毒有效转入靶细胞。用带报告基因LacZ的腺病毒感染大鼠C5脑胶质瘤细胞，经x-gal染色，证明腺病毒载体的基因转移效率可达100％。

A Gene Transfer System Mediated by Adenovirus and Its Effective Expression

The recombinated plasmid pAdCMV-tk,containing the HSV - tk gene controlled by the CMV promoter was constructed successfully,then cotransfected with the main construction of adenovirus 5,pJM17,into the adenovirus pakaging cell 293. Thus a great deal of reconstructal adenovirus AdCMV -tk,which has the infection effect but Cannot reproduce in any cells other than adenovirus producing cells like 293,was produced. After it was used to infect the rat glioma cell C6,we attested the HSV-tk gene by PCR,of Which the result confirmed that the HSV-tk gene couId be effectively transfered into targeting cells by adenovirus. At the same time,we used the identical vector carrying a reporter gene(lacZ) which is easier to use for detecting C6 cells and rat C6 glioma by the same method. The cultured cells and the tumor slide were dyed by means of X-gal,the results indicated that the transfer efficiency could be as high as 100%.