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

Abstract:

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. mori cell lines （Bm-e-HNU5）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 α-tub，IE and Fib enhanced the transient expression activity of RFP reporter gene in the Bm-e-HNU5, and their activity strengthened sequentially.

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Key words: Bombyx mori cells, promoter, RFPreporter gene, transient expression activity, lipoidmediated method