弗氏2a志贺菌CobB和GtrⅡ蛋白水平及毒力相关性的初步研究  

CobB and GtrⅡ of Shigella flexneri 2a:a pilot study

作　　者：刘小青[1,2] 朱力[1] 冯尔玲[1] 马姗姗[1] 魏华[2] 王恒樑[1] 

机构地区：[1]军事医学科学院生物工程研究所病原微生物生物安全国家重点实验室,北京100071 [2]南昌大学食品科学与技术国家重点实验室,南昌330047

出　　处：《军事医学科学院院刊》2010年第4期351-355,共5页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金(30470101;30700035);国家重点基础研究发展计划(973计划)(2005CB522904)

摘　　要：目的研究翻译后修饰基因cobB和gtrⅡ对志贺菌致病能力的影响。方法采用λ-Red重组系统构建福氏2a志贺菌2457T的cobB,gtrⅡ缺失突变株,进行初步的毒力评价,随后制备野生株,cobB,gtrⅡ缺失突变株的全菌蛋白样品,并进行双向电泳比较野生株和缺失株在蛋白表达谱上的差异。结果成功构建cobB、gtrⅡ基因的缺失突变株,豚鼠角膜实验发现cobB、gtrⅡ缺失株症状稍弱于野生株,但在全菌蛋白表达谱中野生株和突变株并无明显差异。结论双向电泳难以有效地呈现CobB和GtrⅡ的翻译后修饰能力。Objective To investigate the function of genes cobB and gtrⅡ in Shigella flexneri 2a.Method With the help of λ-Red recombinant system,cobB and gtrⅡwere individually knocked out from Sh.flexneri 2a 2457T.The evaluation of mutant virulence was performed,and the protein samples of mutants and wild-type strain were prepared and run by two-dimensional electrophoresis（2-DE）.The 2-DE maps of mutants and wild-type strain were compared to find the differentially expressed proteins.Results The mutants of cobB and gtrⅡwere successfully constructed,and their infective symptoms were slightly weaker than those of the wild-type strain by Sereny test.But there was no obvious difference in their proteome profiles.Conclusion The disparity of post-translational modification proteins caused by CobB and GtrⅡ could not be well exhibited by 2-DE.

关 键 词：志贺菌 弗氏 翻译后修饰基因 λ-Red重组系统 基因敲除 

分 类 号：R378.25[医药卫生—病原生物学]