弗氏2a志贺氏菌2457T株yciD基因缺失突变株的构建  

Construction of yciD Deletion Mutant of Shigella flexneri 2a Strain 2457T

作　　者：张影[1] 朱力[1] 袁静[1] 刘先凯[1] 冯尔玲[1] 王恒樑[1] 朱厚础[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2006年第4期483-488,共6页Letters in Biotechnology

基　　金：国家重点基础研究发展规划项目(2005CB522904);国家自然科学基金项目(30470101)

摘　　要：目的:构建弗氏2a志贺氏菌2457T株yciD基因缺失突变体,以研究yciD基因的功能。方法:根据弗氏2a志贺氏菌2457T株基因组全序列,采用Red重组系统对yciD基因进行缺失,并经PCR和SDS-PAGE证实;对野生株和突变株的生长状态及生化反应进行比较研究。结果:构建了弗氏2a志贺氏菌2457T株的yciD基因缺失突变株2457TΔyciD,该突变株外膜蛋白样品中缺失了一条相对分子质量与从yciD基因推导的蛋白相当(约22000)的蛋白带。该突变株比野生株生长快,利用葡萄糖和甘露醇的能力也比野生株大为增强。结论:获得了弗氏2a志贺氏菌2457T株的yciD基因缺失突变株。Objective： To construct a yciD deletion mutant of Shigella flexneri 2a strain 2457T, in order to explore the function of yciD of Sflexneri 2a 2457T. Methods： In according to complete genome sequence of S.flexneri 2a 2457T, the yciD gene was deleted by using Red recombination system, and confirmed by PCR and SDS-PAGE. The growth of mutant and wild strain were measured separately, and some of biochemical events of mutant and wild strain were comparatively investigated. Results： The yciD deletion mutant strain 2457TAyciD of S.flexneri 2a 2457T was constructed. The protein band, which molecular weight was about 22 000 corresponding to sequence deduced from yciD gene, was absent in sample of outer membrane protein. The mutant strain grew faster than wild strain, and their capability to utilize glucose and mannitol was more than wild strain. Conclusion： The yciD deletion mutant strain of S.flexneri 2a 2457T was obtained.

关 键 词：弗氏2a志贺氏菌2457T株 外膜蛋白 RED重组系统 基因敲除 

分 类 号：Q75[生物学—分子生物学] Q78