家蚕蛹期特异基表达因BmCP283鉴定及其启动子的克隆分析

【目的】鉴定家蚕蛹期特异基因及其启动子，为家蚕变态发育人为调节及蛹生物反应器开发提供支撑。【方法】基于芯片表达数据分析及RT-PCR验证，筛选家蚕蛹期特异表达基因；利用PCR方法克隆家蚕蛹期特异表达基因上游的启动子序列，并利用转基因技术验证启动子的活性及时期特异性。【结果】筛选获得了在家蚕蛹期特异表达的表皮蛋白基因BmCP283；所克隆的BmCP283上游启动子区序列长2 004 bp，利用此序列所构建以红色荧光蛋白基因dsRed为报告基因的转基因蚕中，BmCP283启动子驱动的dsRed只在蛹后期表达，且主要表达于翅等组织中。【结论】BmCP283为家蚕蛹期特异表达基因，克隆获得的BmCP283启动子的启动活性具有蛹期特异性。

Identification of Silkworm Pupa-Specific Gene BmCP283 and Its Promoter

【Objective】The objective of this study is to identify genes that are specifically expressed during silkworm pupa-adult transition, clone their promoters and provide supports for controlling artificially pupa-adult transition and develope pupal bioreactor in silkworm (Bombyx mori).【Method】Silkworm pupa-specific genes were identified through analysis of microarray data for gene expression during silkworm metamorphosis and confirmed by RT-PCR experiments. The promoters of silkworm pupa-specific genes were cloned by PCR experiments and their driving activities and pupa-specificity were detected by transgenic technology.【Result】Cuticular protein gene BmCP283 was found to be specifically expressed at pupal stage in silkworm and was regarded as a pupa-specific gene. The potential promoter with a length of 2 004 bp on the upstream of the translational initial site of BmCP283 was cloned. Transgenic experiments confirmed that the promoter of BmCP283 could drive the red fluorescent gene dsRed to be specifically expressed at late pupal stages, which was similar to the expression profile of endogenous BmCP283. 【Conclusion】Both the expression pattern of BmCP283 and the activities of its promoter is pupa-specific in silkworm.