家蚕丝素重链启动子克隆片段在家蚕体内和昆虫培养细胞内的渗漏表达

为了探讨家蚕丝素重链基因的调控机制,应用重组苜蓿银纹夜蛾核型多角体病毒(AcMNPV)为基因转移载体,以绿色荧光蛋白基因为报告基因,研究了丝素重链基因启动子克隆片段在家蚕丝腺的不同部位,以及脂肪体细胞和Sf9培养细胞中的表达渗漏。结果发现,丝素重链启动子克隆片段在家蚕中部丝腺存在表达渗漏,并且在中部丝腺前部的表达活性要强于中部丝腺的中部和后部；克隆片段在脂肪体组织存在表达渗漏,通过荧光解剖显微镜可以在体表观察到绿色荧光；克隆片段在Sf9培养细胞中也存在表达渗漏。上述结果说明目前已知的有关家蚕丝素重链基因的调控元件仍不充分,家蚕基因组中可能还有其他调控丝素重链基因表达组织和时相的特异性元件存在。

Leaked Expression of Cloned Fibroin Heavy Chain Promoter Sequence in Silkworm, Bombyx mori, and Insect Cultured Cells

In order to probe into the regulation mechanism of fibroin gene in silkworm,leaked expression of cloned fibroin heavy chain promoter segment in different part of Bombyx mori silk gland,fat body and Sf9 cul- tured cells was studied by using recombinant AcMNPV as gene transfer vector and EGFP as reporter.The re- sults showed that expression leaking happened in middle silk gland(MSG)of silkworm and the relative activity of FibH promoter in anterior MSG is stronger than in middle MSG and posterior MSG,that expression leaking also happened in silkworm fat body,the fluorescence of EGFP can be observed through silkworm cuticle un- der fluorescent microscopy,and that FibH promoter has leaked expression in Sf9 cultured cells as well.These results showed that current known data on the controlling of FibH is not enough,other factors evolved in con- trolling of FibH may exist in silkworm.