关键词: 家蚕, Ser1基因, 启动子, 活性分析

Abstract: Sericin1 gene (Ser1) of silkworm (Bombyx mori) is specifically expressed in middle silkgland, its promoter plays an important role in silkworm bioreactor. In order to learn polymorphisms of upstream regulation sequence of Ser1 gene from  silkworm breed Yun 7A, we obtained Ser1 promoter sequences that had driver activity, Ser1 promoter sequence was cloned and then analyzed, the expression vector of pBac [Ser1p-DsRed-SV40+A3-EGFP] contained DsRed (red fluorescent protein gene)was driven by Ser1 promoter, the vector was transferred to BmN cells to verify the promoter activity through transient transfection expression assay. Sequence alignment showed that cis element area had been highly conservative between upstream regulation sequence of reported Ser1 and Ser1 promoter sequence of Yun 7A, the other area showed absence of the genetic phenomenon of 422 bp bases; there were conservative sequences of TATA box and CAAT box located at position -24～-30 and -112～-115 respectively, the sequences of TATA box was TATAAAA; the promoter of Ser1 could drive DsRed gene, the transiently expression products could be observed successfully in the cultural BmN cells and middle silkgland. The polymorphism and driver activity of Ser1 upstream regulation sequence from Yunnan silkworm breed were analyzed in this study, so as to lay the foundation for understanding the ability difference of sericin synthetic in the different silkworm breeds and acquiring the efficient promoter.

Key words: Bombyx mori, Ser1 gene, promoter, activity analysis