| pcDNA3与pIRES1.neo筛选克隆效率的比较 |
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| 引用本文： | 谢庆军,王晓彤,陆应麟.pcDNA3与pIRES1.neo筛选克隆效率的比较[J].生物技术通讯,2002,13(2):115-117. |
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| 作者姓名： | 谢庆军 王晓彤 陆应麟 |
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| 作者单位： | 军事医学科学院基础医学研究所,北京100850 |
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| 基金项目： | 全军十五规划重点项目(01Z024)资助 |
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| 摘 要： | 将绿色荧光蛋白(GFP)报告基因分别插入pIRES1.neo与pcDNA3二真核表达载体,构建pGFP-IRES1.neo和pcDNA3-GFP,转染小鼠B16细胞和人HepG2细胞,经G418筛选出抗性克隆,在倒置荧光显微镜下观察,鉴定共表达克隆数;结果在两个细胞系中,pcDNA3-GFP共表达率分别为0%和4%,而pIRES1.neo-GFP共表达率为75%和100%,差异非常显著;因此在筛选稳定表达目的基因细胞克隆的实验中,pIRES1.neo是一个较理想的载体。
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| 关 键 词： | 内部核糖体进入位点序列 绿色荧光蛋白 真核表达载体 克隆筛选效率 |
| 文章编号： | 1009-0002(2002)02-0115-03 |
| 修稿时间： | 2001年10月8日 |
Comparison of two vectors--pcDNA3&pIRESl.neo-in their efficiency of clone screening |
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| XIE Qing-jun,WANG Xiao-tong,LU Ying-lin.Comparison of two vectors--pcDNA3&pIRESl.neo-in their efficiency of clone screening[J].Letters in Biotechnology,2002,13(2):115-117. |
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| Authors: | XIE Qing-jun WANG Xiao-tong LU Ying-lin |
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| Abstract: | Insert the reporter gene green fluorescence protein(GFP)into the two eukaryotic expressing vectors to construct pGFP-IRES1.neo and pcDNA3-GFP,then transfect the mouse B16and human HepG2cell lines,after the administration of G418,resistant clones appears;finally to identify the co-expressing clones under a inverted fluo-rescent microscope.In two cell lines,the co-expressing efficiency of pcDNA3-GFP is0%and4%,whereas,the co-expressing efficiency of pGFP-IRES1.neo is75%and100%respectively.Therefore,pIRES1.neo is an ideal vector to establish a cell line stably expressing an interest gene |
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| Keywords: | internal ribosome entry site sequence green fluorescence protein eukaryotic expressing vector clone screening efficiency |
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