mIL-12重组逆转录病毒载体的构建及包装细胞系的建立  

Construction of mIL-12 recombinant retrovirus vector and establishment of viral package cell line

作　　者：李菁华[1] 陈东[2] 刘颖[2] 

机构地区：[1]吉林大学基础医学院病原生物学教研室,吉林长春130021 [2]吉林大学基础医学院组织学与胚胎学教研室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2006年第2期254-256,共3页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅资助课题(20030542-1);长春市科技计划项目资助课题(04-07SF030)

摘　　要：目的:构建mIL-12重组逆转录病毒载体。方法:将mIL-12 DNA克隆到载体pLEGFP,用该重组质粒转染PA317包装细胞获得重组逆转录病毒。结果:重组质粒pLEGFP-mIL12酶切及PCR产物在2 290 bp区域见强荧光条带与mIL-12基因大小相符;转染PA317细胞后的上清感染NIH3T3细胞后,经共聚焦显微镜能观察到融合蛋白的表达。结论:包装细胞上清中有含mIL-12 DNA的重组逆转录病毒,mIL-12重组逆转录病毒可感染NIH3T3细胞并在细胞中进行有效的复制。Objective To construct mIL-12 recombinant retrovirus vector and evaluate the effect of mIL-12 expression on glioma in rats. Methods mIL-12 DNA was cloned into vector pLEGFP using standard procedures to develop recombinant plasmid pLEGFP-mIL-12 , then it was transferred into PA317 cells. Results Products of enzyme digestion and PCR of recombinant plasmid pLEGFP-mIL12 were analysed using electrophoresis and a 2 290 bp fragment of DNA as same as mIL-12 gene in size was found, transfected NIH3T3 expressed GFP-mIL12 fusion protein. Conclusion Transfected PA317 cells can produce retrovirus which can infect NIH3T3 cells, and the expression of mIL-12 gene can be detected in NIH3T3 cells.

关 键 词：MIL-12基因 逆转录病毒载体 包装细胞 

分 类 号：Q78[生物学—分子生物学]