›› 2006, Vol. 49 ›› Issue (2): 167-171.

• RESEARCH PAPERS •     Next Articles

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

WANG Yun,YE Xiang-Qun, WU Yi-Liang, GUI Mu-Yan, ZUO Zheng-Hong   

  • Online:2006-05-12 Published:2006-11-20

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.


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