mIL-12重组逆转录病毒载体的构建及包装细胞系的建立

doi:吉林省科技厅资助课题（20030542-1）；长春

mIL-12重组逆转录病毒载体的构建及包装细胞系的建立

李菁华¹，陈东^2*，刘颖²

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

收稿日期:2005-04-07 修回日期:1900-01-01 出版日期:2006-03-28 发布日期:2006-03-28
通讯作者: 陈东^2*

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

LI Jing-hua¹,LI Jing-hua¹, CHEN Dong^2*，LIU Ying²LIU Ying²

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

Received:2005-04-07 Revised:1900-01-01 Online:2006-03-28 Published:2006-03-28
Contact: CHEN Dong^2*

摘要/Abstract

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: 逆转录病毒载体, 包装细胞

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. ls can produce retrovirus which can infect NIH3T3 cells,and the expression of mIL-12 gene can be detected in NIH3T3 cells.

Key words: retroviridae, package cell

中图分类号:

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]. J4, 2006, 32(2): 254-256.

LI Jing-hua,LI Jing-hua, CHEN Dong，LIU YingLIU Ying. Construction of mIL-12 recombinant retrovirus vector and establishment of viral package cell line[J]. J4, 2006, 32(2): 254-256.

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