The Luciferase (luc) reporter gene was used to construct recombinant plasmids containing six different promoters, which are the cytoplasmic actin3 promoter (A3), the fibroin heavy chain promoter (Fib-H), the fibroin light chain promoter (Fib-L), a glycoprotein (P25) promoter, Sericin-1 promoter (Ser-1) and the Bombyx mori nuclear polyhedrosis virus immediate (BmNPV) early protein promoter 1(IE-1), respectively. These recombinant plasmids, which are pGL3-A3-luc, pGL3-Fib-H-luc, pGL3-Fib-L-luc, pGL3-P25-luc, pGL3-Ser-1-luc and pGL3-IE-1-luc had been constructed successfully by restriction enzyme digestion and PCR analysis, and then were transfected into BmN cells to measure the activity of the six promoters to drive luc reporter gene transient expression in cells. Transfection and transcriptional experiment indicated that except pGL3-Fib-L-luc, pGL3-P25-luc, pGL3-Ser-1-luc, others three kinds of recombinant plasmids all transfected BmN cells obviously. The promoters of Fib-H, A3 and IE-1 enhanced the transient expression activity of luc reporter gene in the BmN cells and their activity strengthened sequentially.
Russian Journal of Developmental Biology – Springer Journals
Published: Nov 16, 2012
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