摘要
利用逆转录病毒载体pLNSX构建了带HSVTK基因的逆转录病毒重组体pLNSTK,用磷酸钙沉淀法转染PA317包装细胞,经G418筛选抗性克隆,建立了产生重组病毒的载体产生细胞系PA317/TK,用NIH3T3细胞测定病毒(CFU)滴度为3×108L-1。提取PA317/TK细胞的DNA和细胞上清液的RNA,在特异性引物引导下,分别用PCR和RTPCR检测证实PA317/TK细胞整合了HSVTK基因并产生重组病毒,为HSVTK基因治疗恶性肿瘤的实验研究打下了基础。
he recombinant retrovirus expressive vector pLNS TK was constructed with HSV TK cDNA and retrovirus vector pLNSX. The pLNS TK was certified by restriction endonuclease digestion and transfected into PA317 packing cell by calcium phosphate transfection method. The HSV TK vector producer cell line PA317/TK was selected in G418 medium. The virus titer was about 3×10 8 CFU/L with NIH3T3 cells. The integration of HSV TK cDNA into PA317 producing cells was identified by PCR method in the genomic DNA and by RT PCR method in RNA of supernatant of PA317/TK cells. The experiment makes it possible to carry out tumor gene therapy with HSV TK gene.
出处
《中山医科大学学报》
CSCD
1997年第4期272-274,共3页
Academic Journal of Sun Yat-sen University of Medical Sciences
基金
中山医科大学科研基金
关键词
胸苷激酶
逆转录病毒
重组
单纯疱疹病毒
simplex virus/genetics
thymidine kinase/genetics
retroviride/genetics
recombination, genetic