| 重组逆转录病毒载体的包装 |
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| 引用本文： | 王凤阳,扈荣良,张茂林,赵君,涂长春,殷震.重组逆转录病毒载体的包装[J].中国兽医学报,2001,21(2):128-130. |
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| 作者姓名： | 王凤阳 扈荣良 张茂林 赵君 涂长春 殷震 |
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| 作者单位： | 解放军军需大学军事兽医研究所, |
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| 基金项目： | 国家“863”计划重大资助项目(Z21-04-03) |
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| 摘 要： | 将3.7kb的LacZ基因片段克隆到含标志基因Neo的逆转录病毒载体，pLXSN与pLNCX的多克隆位点，构建了含有LacZ基因的重组逆转录病毒载体pLLSN和pLNCL.在测定了G418对饥装细胞系PA317的最小致剂量后（最小致死剂量为0.3g/L），通过脂质体Lipofectin与磷酸钙2种介质，分别将2种重组逆转录病毒载体导入包装细胞系PA317进行包装，并以0.3g/L，的G418进行筛选，得到多个G418抗性克隆，经扩大培养，传代，分别得到包装pLLSN与pLNCL的2株阳性细胞，电镜下可见阳性细胞的培养上清中具有逆转录病毒形态特征的成熟病毒粒子，本0研究为运用逆转录病毒载体系统，解决转角因效率低的问题奠定了基础。
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| 关 键 词： | 重组逆转录病毒载体 包装细胞系 基因转移 包装 转基因效率 |
| 文章编号： | 1005-4545(2001)02-0128-03 |
| 修稿时间： | 2000年3月26日 |
Packaging of Recombinant Retroviral Vectors in Vitro |
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| WANG Feng-yang,HU Rong-liang ,ZHANG Mao-lin,ZHAO Jun,TU Chang-chun,YIN Zhen.Packaging of Recombinant Retroviral Vectors in Vitro[J].Chinese Journal of Veterinary Science,2001,21(2):128-130. |
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| Authors: | WANG Feng-yang HU Rong-liang ZHANG Mao-lin ZHAO Jun TU Chang-chun YIN Zhen |
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| Institution: | WANG Feng-yang,HU Rong-liang *,ZHANG Mao-lin,ZHAO Jun,TU Chang-chun,YIN Zhen |
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| Abstract: | Recombinant retroviral vectors, pLLSN and pLNCL carrying LacZ have been constructed.The two vectors were transfected into PA317 for packaging by Lipofectin and calcium phosphat precipitation.After G418 selection at a concentration of 0.3 g/L for 2 weeks ,G418-resistant PA317 colonies were obtained,and then they were amplified.The supernatants of the cell cultures were harvested, concentracted and oberserved under transmission electron microscope, the mature virus particles possessed the typically morphological feature of retrovirus have been found. |
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| Keywords: | recombinant retroviral vetors LacZ packaging |
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