家蚕蛹期特异基因BmCP283的启动子活性分析  

Promoter Activity Analysis of Bombyx mori Pupal Stage Specific Gene BmCP283

作　　者：康丽霞[1] 孟勐[1] 王永虎[1] 唐林[1] 彭健[1] 钱文良[1] 程道军[1] 夏庆友[1] 

机构地区：[1]家蚕基因组生物学国家重点实验室,西南大学,重庆400715

出　　处：《蚕业科学》2013年第5期881-886,共6页ACTA SERICOLOGICA SINICA

基　　金：国家重点基础研究发展计划“973”项目(No.2012CB1146-00);国家高技术研究发展计划“863”项目(No.2011AA-100306);国家自然科学基金项目(No.31272503);重庆市自然科学基金项目(No.cstc2012jjA80023)

摘　　要：前期研究鉴定了家蚕蛹期特异表达基因BmCP283,并克隆了翻译起始位点上游长度为2 004 bp的启动子区序列BmCP283P。用PCR方法扩增了BmCP283P启动子9个不同区域的片段,构建由其驱动的荧光素酶基因报告载体,并转染BmE细胞进一步分析该启动子的活性。结果显示,BmCP283P启动子序列由上游2 004 bp递减至1 743 bp,以及由1 608 bp递减至1 328 bp,其活性显著增强;而从1 328 bp递减至1 189 bp,其活性则显著减弱。这暗示2 004～1 743 bp和1 608～1 328 bp区域可能存在抑制BmCP283P启动子活性的调控元件,而1 328～1 189 bp区域可能存在启动子活性增强元件。另外,蜕皮激素(20E)诱导能增强BmCP283P启动子的活性,暗示20E参与了BmCP283基因的转录调控。研究结果不仅有助于阐明BmCP283基因的转录调控机理,也为人为调控家蚕蛹期发育提供了潜在的可用启动子。In previous report, we indentified a pupal stage specific gene BmCP283 in silkworm （Bombyx mori） and cloned its promoter sequence BmCP283P covering the upstream 2 004 bp of its translational initiation site. Herein, we amplified nine different fragments from BmCP283P by PCR, constructed recombinant vectors carrying luciferase reporter gene driven by these fragments, and transformed them into BmE cells for further analysis on activity of promoter Bm- CP283P. The results showed that the activity of promoter BmCP283P significantly increased when the upstream se- quence was shortened from 2 004 bp to 1 743 bp and from 1 608 to 1 328 bp, but significantly declined when it was shortened from 1 328 bp to 1 189 bp. It was suggested that the regions from 2 004 to 1 743 bp and from 1 608 to 1 328 bp contained elements for binding activators to inhibit promoter activity of BmCP283P, whereas the region from 1 328 to 1 189 bp contained elements for binding enhancers. Moreover, the activity of promoter BmCP283P could be increased upon induction by 20-hydroxyecdysone （20E）, suggesting that 20E was involved in the transcriptional regulation of Bm-CP283 gene. Our results not only facilitate further elucid- ation of transcriptional regulatory mechanism of BmCP283 gene but also provide potential usable promoters for artifi- cial regulation of silkworm pupa development.

关 键 词：家蚕 BmCP283基因 启动子 活性区域 蜕皮激素 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q782[农业科学—畜牧兽医]