四种启动子调控RFP报告基因在家蚕细胞(Bm-e-HNU5)内的瞬时表达  

Comparison of four promoters for transient expression of RFP reporter gene in cultured Bombyx mori cells (Bm-e-HNU5)

作　　者：王云[1] 叶向群[1] 吴亦亮[1] 桂慕燕[1] 左正宏[1] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005

出　　处：《昆虫学报》2006年第2期167-171,共5页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(39870410)

摘　　要：以红色荧光蛋白基因(RFP)为报告基因,构建含4种不同启动子的重组表达质粒,用脂质体介导法转染家蚕Bombyxmori细胞(Bm-e-HNU5),观察家蚕细胞质肌动蛋白4基因启动子(A4)、α微管蛋白基因启动子(α-tub)、蚕丝心蛋白重链基因启动子(Fib)和家蚕核型多角体病毒早期即刻蛋白基因启动子(IE)4种启动子调控RFP报告基因在家蚕细胞内的瞬时表达情况。构建的重组表达质粒pDsRed-α-tub、pDsRed-A4、pDsRed-IE和pDsRed-Fib经双酶切和PCR鉴定正确无误。转染和转录实验结果表明,除了pDsRed-A4外,其他3种重组质粒在Bm-e-HNU5细胞中都得到高转染率,α-tub、IE和Fib可依次增强RFP报告基因在家蚕细胞内的瞬时表达活性。The red fluorescent protein reporter gene （RFP） was used to construct recombinant plasmids containing four different promoters, i. e. , the cytoplasmic actin4 promoter （A4）, α-tubulin promoter （α-tub） from silkworm, the Bombyx mori nuclear polyhedrosis virus immediate early protein promoter （IE） and the fibroin heavy chain gene promoter （Fib）, respectively. These recombinant plasmids, i. e., pDsRed-A4, pDsRed-α-tub, pDsRed-Fib and pDsRed-IE, had been constructed successfully by restriction enzyme digestion and PCR analysis, and then were transfected into B. mor/ cell lines （Bm-e-HNUS） by lipid-mediated method to observe the ability of the four promoters to drive RFP reporter gene transient expression in cells. Transfection and transcription experiments indicated that except pDsRed-A4, the other three kinds of recombinant plasmids all transfected Bm-e-HNU5 obviously. The promoters of a-tub, IE and Fib enhanced the transient expression activity of RFP reporter gene in the Bm-e-HNUS, and their activity strengthened sequentially.

关 键 词：家蚕 启动子 RFP报告基因 瞬时表达活性 脂质体介导法 

分 类 号：Q966[生物学—昆虫学]