摘要
目的 :选择合适的载体系统用于CS3菌毛的高水平表达和装配。方法 :将CS3操纵子基因分别克隆入pUC19,pTrc99A等载体中 ,采用全细胞ELISA方法检测CS3菌毛的表达水平 ,并用SDS PAGE和电子显微镜技术证实菌毛的表达和组装。结果 :CS3菌毛在质粒pUC19中的表达水平明显高于在pTrc99A和pBR32 2中的表达水平 ,基因的插入方向对表达影响甚微 ;SDS PAGE可看到表达条带 ,电镜下可观察到菌毛形成。结论 :CS3自身的启动子在表达中起决定作用 。
Objective:To choose the best vector for the expression of CS3 fimbriae. Methods: The CS3 operon was cloned into different plasmid vectors such as pUC19 and pTrc99A. The expression of CS3 was monitored by whole cell ELISA and SDS PAGE analysis. The assembly of CS3 fimbriae was detected with electron microscopy. Results:The expression level of CS3 fimbriae using plasmid pUC19 as carrying vector was the highest, and the insertion orientation of CS3 gene into the plasmid has a little effect on its expression level. The expression of CS3 fimbriae was confirmed by SDS PAGE analysis and electron microscopy.Conclusions:The promotor of CS3 itself played the key role in the expression of CS3 fimbriae and the copy number of plasmid was the main factor to affect the expression level. [
出处
《军事医学科学院院刊》
CSCD
北大核心
2001年第1期1-4,共4页
Bulletin of the Academy of Military Medical Sciences
基金
国家自然科学基金资助! (395 70 40 8)
关键词
CS3菌
载体
表达
启动子
CS3 fimbriae
vector
expression
promotor
