家蚕BmRYBP基因的克隆和转基因干涉品系的建立.pdfVIP

国蚕学会第七届青年学术研讨会论文集 家蚕BmR YBP 基因的克隆及转基因干涉品系的建立 1 2 1 2 2 张美蓉 徐汉福 许平震 邓党军 夏庆友 1 2 （中国农业科学院蚕业研究所，江苏镇江 212018； 西南大学蚕学与系统生物学研究所，重庆 400716） 摘 要 Polycomb group 蛋白一群对任何有机体的生长发育都很重要的普遍的转录抑制因子，参与 了几乎所有生物有机体的发育调控。虽然其成员通常在结构域和结构基序上并不相关，却由于行使 共同的功能而形成一个家族，在物种间高度保守。RYBP 基因是小鼠和果蝇 已经证实的PcG 蛋白 成员。本研究基于生物信息学分析结果，克隆了家蚕的RYBP 基因，研究结果显示，该基因在家蚕 五龄3 天幼虫的生殖腺 有最高的表达量。基于GAL4/UAS 系统，我们构建了具有BmR YBP 序列反 向重复结构的转基因表达载体，获得了UAS 品系的转基因后代，在杂交后代 实现了对目标基因的 特异干涉，为研究BmR YBP 在家蚕生长发育过程 行使的功能提供了基础。 关键词 家蚕；BmR YBP ；GAL4/UAS 系统；转基因干涉表达载体；显微注射 Abstract Polycomb group(PcG), a group of very important genes, maintain this established molecular mechanism in life. Although the members of PcG proteins have differences in structure and motifs, they form a common family due to perform the common function. RYBP is a protein reported both in Drosophila melanogaster and Mus musculus, which can interact with other PcG member, and predicted to be a member of PcG protein family. The results of RT-PCR by which to investigate the expressions of rd R YBP of silkworm on 3 day fifth instar indicate that the R YBP expressed almost in all tissues with a high expression levels gonad and head and a lower expression levels in other tissues. We have constructed the IR transgenic expression construct pBac[UASR YBP SV40-3xp3EGFP], and by the strandard method of IR transgenic inj ect we have gotten the UAS-R YBP transgenic silkworm and established the separate silkworm transgenic lines. Key words：Bombyx mori; BmR YBP; the GAL4/UAS system; transgenic expression construct Transgenic RNAi