| mIL-12重组逆转录病毒载体的构建及包装细胞系的建立 |
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| 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. |
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| 作者姓名： | 李菁华 陈东 刘颖 |
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| 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 |
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| 基金项目： | 吉林省科技厅科研项目
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吉林省长春市科技基金 |
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| 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。
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| 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年4月7日 |
Construction of mIL-12 recombinant retrovirus vector and establishment of viral package cell line |
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| 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. |
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| Authors: | LI Jing-hua CHEN Dong LIU Ying |
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| Institution: | 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 |
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| 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. |
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| Keywords: | mIL-12 gene retroviridae package cell |
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