巨细胞病毒和猴空泡病毒40启动子在中国仓鼠卵巢细胞中驱动基因表达效率比较  

Comparison of efficiency of cytomegalovirus and Simian vacuolating virus 40 promoter driving gene expression in Chinese hamster ovary cells

作　　者：王喜成[1] 贾岩龙[1] 郭潇[1] 张玺[2] 王天云[2] 

机构地区：[1]新乡医学院药学院,河南新乡453003 [2]新乡医学院基础医学院,河南新乡453003

出　　处：《新乡医学院学报》2016年第7期553-555,共3页Journal of Xinxiang Medical University

基　　金：国家自然科学基金资助项目(编号:31371332)

摘　　要：目的比较巨细胞病毒(CMV)和猴空泡病毒40(SV40)2种启动子驱动外源基因在中国仓鼠卵巢(CHO)细胞CHO-K1中的表达效率。方法分别构建由CMV和SV40启动子驱动表达绿色荧光蛋白(EGFP)的真核表达载体pWTY-02和pWTY-03。用阳离子脂质体LipofectamineTM2000将表达载体质粒分别转染CHO-K1细胞,倒置荧光显微镜观察转染后CHO-K1细胞中EGFP的表达情况;流式细胞术检测EGFP的荧光强度,以比较2种启动子驱动外源基因表达的效率。结果 2种启动子均能够驱动EGFP在CHO-K1细胞中表达。转染质粒pWTY-02的CHOK1细胞的平均荧光强度明显高于转染质粒pWTY-03的CHO-K1细胞(P<0.05)。结论 CMV启动子在CHO-K1细胞中驱动EGFP的表达效率较高,为后续CHO表达系统中提高外源基因表达效率启动子的选择提供了依据。Objective To compare the efficiency of two kinds of promoters[cytomegalovirus（ CMV） and Simian vacuolating virus 40（ SV40） promoter] driving gene expression in Chinese hamster ovary（ CHO） cells CHO-K1. Methods Eukaryotic expression vectors pWTY-02 and pWTY-03 containing enhanced green fluorescent protein（ EGFP） gene respectively driven by CMV and SV40 promoter were constructed. The vectors were respectively transfected into CHO-K1 cells using lipofectamineTM2000. Expression of EGFP was observed in the transfected CHO-K1 cells under the fluorescence microscope. Fluorescence intensity of EGFP was detected by flow cytometry to compare the efficiency of CMV and SV40 promoter driving gene expression. Results The EGFP expression was detected both in CHO-K1 cells transfected with pWTY-02（ driven by the CMV promoter） and pWTY-03（ driven by the SV40 promoter）. The results of flow cytometry analysis showed that the efficiency of EGFP expression in CHO-K1 cells transfected with pWTY-02 was higher than that with pWTY-03（ P 0. 05）. Conclusions The efficiency of EGFP expression driven by the CMV promoter was higher than the SV40 promoter in the transfected CHOK1 cells. The findings provide theoretical basis on application of promoter for CHO expression system.

关 键 词：巨细胞病毒启动子 猴空泡病毒40启动子 绿色荧光蛋白 中国仓鼠卵巢细胞 

分 类 号：Q789[生物学—分子生物学]