含HSV-TK基因及IL-18基因复制缺陷型腺病毒载体的构建和鉴定  

Construction and Identification of Replication-deficient Adenovirus Vector Containing TK Gene and IL-18 Gene

作　　者：徐春华[1] 刘越[1] 肖利民[1] 邓圣泽[1] 李东海[1] 

机构地区：[1]南昌大学第一附属医院,江西南昌330006

出　　处：《中国医学创新》2014年第36期13-15,共3页Medical Innovation of China

摘　　要：目的:分别构建含TK及IL-18基因的重组腺病毒载体,并进行鉴定。方法:通过DNA重组技术,构建含有目的基因的复制缺陷型腺病毒Ad CMV-TK及Ad CMV-IL-18;将上述重组腺病毒液感染293细胞,提取DNA,进行PCR鉴定;将病毒液感染293细胞,扩增病毒,用OD260法测定病毒滴度。结果:经PCR鉴定,鉴定正确的腺病毒命名为Ad CMV-TK及Ad CMV-IL-18,测定病毒滴度Ad CMV-TK=1.28×109 PFU/m L;Ad CMV-IL-18=1.28×109 PFU/m L。结论:成功构建了含HSV-TK基因及IL-18基因重组腺病毒载体,为肿瘤的基因治疗提供了理论依据及实验数据。Objective：To construct and identify the recombinant adenovirus vector containing TK gene and IL-18 gene resepectively. Method： Replication-deficient adenovirus vector containing Ad CMV-TK gene and Ad CMV-IL-18 gene were constructed by using DNA recombination;293 cells were transduced by Ad CMV-TK gene and Ad CMVIL-18 gene,then DNA of those transduced cells was extracted and PCR was conducted; Ad CMV-TK and Ad CMVIL-18 recombinant adenovirus were efficently amplified in 293 cells and the titer of the virus was determined by OD260 assay. Result：the Ad CMV-TK gene and Ad CMV-IL-18 gene were identified correctly by PCR,the titer of the virus：Ad CMV-TK=1.28×10^9 PFU/m L;Ad CMV-IL-18=1.28×10^9 PFU/m L.Conclusion： Replication-deficient Recombinant Adenovirus Vector Containing HSV-TK gene and IL-18 gene are constructed successfully, it would provide some basic theory and experimental data for the tumor gene therapy.

关 键 词：TK基因 IL-18基因 腺病毒载体 构建 鉴定 

分 类 号：R346[医药卫生—基础医学]