家蚕蛹期特异基表达因BmCP283鉴定及其启动子的克隆分析.pdfVIP

中国农业科学 2013,46(11):2353-2362 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2013.11.020 家蚕蛹期特异基表达因BmCP283 鉴定及其启动子的克隆分析 程道军，唐 林，孟 勐，康丽霞，王永虎，彭 健，马三垣，夏庆友 （西南大学家蚕基因组生物学国家重点实验室，重庆 400715 ） 摘要：【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。 关键词：家蚕；蛹期；启动子；特异；转基因 Identification of Silkworm Pupa-Specific Gene BmCP283 and Its Promoter CHENG Dao-jun, TANG Lin, MENG Meng, KANG Li -xia, WANG Yong-hu, PENG Jian, MA San-yuan, XIA Qing-you (State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715) Abstract: 【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 translatio