李桂英, 孙红光, 王磊, 田宇, 杜柏榕, 朱迅. L-plastin启动子调控下的肿瘤细胞特异性表达载体的构建及活性分析[J]. 中国肿瘤临床, 2007, 34(17): 966-968,969.
引用本文: 李桂英, 孙红光, 王磊, 田宇, 杜柏榕, 朱迅. L-plastin启动子调控下的肿瘤细胞特异性表达载体的构建及活性分析[J]. 中国肿瘤临床, 2007, 34(17): 966-968,969.
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Li Guiying, Sun Hongguang, Wang Lei, Tian Yu, Du Borong, Zhu Xun. Construction and Functional Analysis of a Tumor Cell-Specific Expression Vector Regulated by the L-plastin Promoter[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2007, 34(17): 966-968,969.
Citation: Li Guiying, Sun Hongguang, Wang Lei, Tian Yu, Du Borong, Zhu Xun. Construction and Functional Analysis of a Tumor Cell-Specific Expression Vector Regulated by the L-plastin Promoter[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2007, 34(17): 966-968,969.
shu

L-plastin启动子调控下的肿瘤细胞特异性表达载体的构建及活性分析

Construction and Functional Analysis of a Tumor Cell-Specific Expression Vector Regulated by the L-plastin Promoter

    摘要
  • 摘要: 目的 :构建肿瘤细胞特异性表达载体pcDNA3.1PLN-GFP及鉴定活性。 方法 :用2.4kb的L-plastin启动子,以绿色荧光蛋白(GFP)为报告蛋白,通过分子克隆技术,构建真核表达载体pcDNA3.1PLN-GFP,用脂质体介导法转染肿瘤细胞和成纤维细胞后,流式细胞术(FACS)分析载体的肿瘤细胞特异性和活性。 结果 :仅在表达内源性L-plastin的肿瘤细胞和转化的293细胞中有GFP的表达,发光细胞比率较高,而且L-plastin启动子的活性与CMV启动子活性相当,而在其它细胞中没有检测到GFP的表达。 结论 :我们构建的pcDNA3.1PLN-GFP是一肿瘤特异性的高效表达载体。

     

    Abstract: Objective :To construct the tumor cell-specific expression vector pcDNA3.1PLN-GFPand analyze its activity. Methods :The 2.4 kb L-plastin promoter was used as a promoter and greenfluorescent protein (GFP) was used as reporter to construct the expression vector pcDNA3.1PLN-GFPwith molecular cloning techniques.The recombinant plasmid was transfected into tumor cells andmurine fibroblasts with lipofectin. The tumor specificity and activity of this vector were analyzed by flu-orescence activated cell sorting. Results :L-plastin GFP protein was expressed only in tumor cells andtransformed HEK293 cells that expressed the endogenous L-plastin gene.Its activity was equivalent tothat found with a vector containing the CMV promoter. Conclusion :The pcDNA3.1PLN-GFP vector isan efficient tumor cell-specific expression vector.

     

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