家蚕丝素P25蛋白基因启动子顺式作用元件的功能分析  

Functional analysis of cis-acting elements of promoter of fibroin P25 gene in the silkworm,Bombyx mori

作　　者：汪生鹏[1,2] 孙霞[2] 沈小绢[2] 彭伟[2] 郭锡杰[1,2] 

机构地区：[1]中国农业科学院蚕业研究所农业部蚕桑遗传改良重点开放实验室,江苏镇江212018 [2]江苏科技大学生物与环境工程学院,江苏镇江212018

出　　处：《昆虫学报》2011年第6期623-633,共11页Acta Entomologica Sinica

基　　金：国家重点基础性研究发展规划("973"计划)项目(2005CB121004);国家自然科学基金项目(30771631);江苏省自然科学基金项目(BK2007099);江苏省普通高校研究生科研创新计划(CX09S_001Z)

摘　　要：为了阐明蚕丝蛋白基因表达调控的分子机制,利用Bac-to-Bac杆状病毒表达系统及实时荧光定量PCR等技术,对家蚕Bombyx mori丝素P25基因启动子上游1233bp的活性及其调控元件的功能进行了分析。结果表明:在P25启动子上游-423～-1233区和-127～-238区存在正调控元件,在-238～-423区段存在负调控元件;PSGF和BMFA两结合元件在P25基因表达中起负调控作用。PSGF结合元件对A3启动子在后部丝腺的活性具有一定的增强作用,进一步验证了PSGF调控元件的功能。通过对P25基因启动子的活性分析,尤其是对PSGF和BMFA调控元件的功能分析,有利于进一步了解P25基因表达调控的精细机制。To study the regulation mechanism of silk genes in the silkworm,Bombyx mori,the upstream 1 233 bp promoter sequence of fibroin P25 gene and its regulatory elements were analyzed by using Bac-to-Bac expression system and real-time quantitative PCR technique.The results showed that there existed positive regulatory elements at position-423 to-1 233 and-127 to-238 in P25 promoter region,and negative regulatory elements at position-238 to-423.PSGF and BMFA regulatory elements played a negative regulation role in P25 expression.PSGF element had a certain degree of enhancement to A3 promoter＇s activity in posterior silk glands,which further validates the function of PSGF regulatory element.Analysis of P25 promoter activity,especially functional analysis of PSGF and BMFA regulatory elements,could be helpful in further understanding the mechanism of P25 expression and fine regulation.

关 键 词：家蚕 丝素P25蛋白 启动子 顺式作用元件 BAC-TO-BAC 实时定量PCR 

分 类 号：Q966[生物学—昆虫学]