含HSVtk基因重组逆转录病毒包装细胞PA317/TK的建立  

Construction of the package cells( PA317/TK) with recombinant retroviral vector containing HSVtk gene

作　　者：陈道桢[1] 李淑锋[1] 鲁晓萱[1] 王静[1] 苏宁[2] 刘璐[3] 黄鹰[3] 童冠圣[4] 洪泽辉[1] 张丽珊[1] 

机构地区：[1]东南大学医学院遗传中心,江苏南京210009 [2]东南大学医学院病理教研室,江苏南京210009 [3]东南大学医学院放免中心,江苏南京210009 [4]铁道部北京铁路总医院核医学科,北京100038

出　　处：《医学研究生学报》2003年第1期1-3,共3页Journal of Medical Postgraduates

基　　金：国家自然科学基金资助课题 (批准号 :3 0 0 70 2 2 9)

摘　　要：目的 :研究肿瘤的自杀基因疗法 ,建立含单纯疱疹病毒胸苷激酶 (HSVtk)基因重组逆转录病毒包装细胞PA317/TK。　方法 :应用脂质体转移方法将逆转录病毒载体GINaTK导入PA317,经G4 18筛选阳性克隆细胞后分别用PCR和扫描电镜检测HSVtk基因整合和病毒颗粒分泌情况。　结果 :逆转录病毒载体GINaTK导入PA317,G4 18后筛选得到阳性克隆细胞 ,命名为PA317/TK。PCR检测PA317/TK细胞出现一特异性 4 0 4bp阳性条带 ,而PA317细胞没有扩增出相应条带。PA317/TK细胞经扫描电镜检测见其表面有颗粒状突起 ,其周围有许多呈团状的病毒颗粒。证实含HSVtk基因重组逆转录病毒存在于包装细胞中。　结论Objectives:In order to study suicide gene therapy on tumor, we constructed the package cell PA317/TK with recombinant retroviral vector containing HSVtk gene. Methods:GINaTK retroviral vector containing HSVtk gene was transduced into PA317 retroviral packaging cell by lipofectin,PA317 cells transfected with HSVtk gene were then screened by G418 medium,and positive clones were respectively measured with PCR and with scan electron microscrope. Results: PA317 cells were transfected successfully with HSVtk gene by lipofectin(named PA317/TK).HSVtk gene integrated into PA317 cells was further identified with PCR.Package cells contain recombinant retrovirus particulate with HSVtk gene with scan electron microscrope. Conclusions: The package cells( PA317/TK) with recombinant retroviral vector containing HSVtk gene were successfully constructed.

关 键 词：HSVtk 基因重组 逆转录病毒 包装细胞 单纯疱疹病毒胸苷激酶基因 

分 类 号：R752.11[医药卫生—皮肤病学与性病学]