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mIL-12重组逆转录病毒载体的构建及包装细胞系的建立
引用本文:李菁华,陈东,刘颖. mIL-12重组逆转录病毒载体的构建及包装细胞系的建立[J]. 吉林大学学报(医学版), 2006, 32(2): 254-256. DOI: 吉林省科技厅资助课题(20030542-1);长春
作者姓名:李菁华  陈东  刘颖
作者单位:吉林大学基础医学院病原生物学教研室,吉林,长春,130021;吉林大学基础医学院组织学与胚胎学教研室,吉林,长春,130021
基金项目:吉林省科技厅科研项目 , 吉林省长春市科技基金
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。

关 键 词:mIL-12基因  逆转录病毒载体  包装细胞
文章编号:1671-587X(2006)02-0254-03
收稿时间:2005-04-07
修稿时间:2005-04-07

Construction of mIL-12 recombinant retrovirus vector and establishment of viral package cell line
LI Jing-hua,CHEN Dong,LIU Ying. Construction of mIL-12 recombinant retrovirus vector and establishment of viral package cell line[J]. Journal of Jilin University: Med Ed, 2006, 32(2): 254-256. DOI: 吉林省科技厅资助课题(20030542-1);长春
Authors:LI Jing-hua  CHEN Dong  LIU Ying
Affiliation:1.Department of Pathogenobiology, School of Basic Medical Sciences,Jilin University,Changchun 130021,China;2. Department of Histology and Embryology, School of Basic Medical Sciences,Jilin University,Changchun 130021,China
Abstract:
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.
Keywords:mIL-12 gene  retroviridae  package cell
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