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| 弗氏志贺菌2a GadB蛋白的功能研究 | |
| 引用本文： | 赵天,朱力,冯尔玲,马姗姗,廖祥儒,王恒樑. 弗氏志贺菌2a GadB蛋白的功能研究[J]. 军事医学科学院院刊, 2011, 35(12): 921-927 |
| 作者姓名： | 赵天 朱力 冯尔玲 马姗姗 廖祥儒 王恒樑 |
| 作者单位： | 1. 江南大学工业生物技术教育部重点实验室,江苏无锡,214122;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. 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 3. 江南大学工业生物技术教育部重点实验室,江苏无锡,214122 |
| 基金项目： | 国家自然科学基金资助项目(810713248,1171531); 国家973计划(2011CB504901) |
| 摘 要： | 目的探究gadB基因的潜在功能及其对弗氏志贺菌2a 301株致病能力的影响。方法利用λ-Red重组系统构建弗氏志贺菌2a 301株的gadB缺失突变株,随后进行基本的表型实验测评,并初步评价其毒力,最后制备突变株和野生株全菌蛋白样品,利用双向电泳技术比较二者在蛋白表达谱上的差异。结果成功获得gadB基因缺失株;生化实验发现缺失株不能利用甘油发酵;而豚鼠角膜实验发现突变株炎症反应稍轻于野生株;全菌蛋白表达谱中发现10个差异蛋白。结论利用双向电泳鉴定到一些可能与GadB有关的蛋白,为gadB潜在功能的研究提供了线索。 |
| 关 键 词： | 志贺菌,弗氏 GadB蛋白 λ-Red重组系统 基因敲除 双向电泳 |
GadB of Shigella flexneri 2a:a pilot study |
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| ZHAO Tian,ZHU Li,FENG Er-ling,MA Shan-shan,LIAO Xiang-ru,WANG Heng-liang. GadB of Shigella flexneri 2a:a pilot study[J]. Bulletin of the Academy of Military Medical Sciences, 2011, 35(12): 921-927 | |
| Authors: | ZHAO Tian ZHU Li FENG Er-ling MA Shan-shan LIAO Xiang-ru WANG Heng-liang |
| Affiliation: | ZHAO Tian1,2,ZHU Li2,FENG Er-ling2,MA Shan-shan2,LIAO Xiang-ru1,WANG Heng-liang1,2 (1.Key Laboratory of Industrial Biotechnology of Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi,Jiangsu 214122,China,2.State Key Laboratory of Pathogen and Biosecurity,Beijing Insitute of Biotechnology,Beijing 100071,China) |
| Abstract: | Objective To investigate the potential functions of gadB in Shigella flexneri 2a.Methods Gene gadB was knocked out from S.flexneri 2a stain 301 by λ-Red recombinant system.The biochemical phenotypes and the virulence of the mutant strain was tested.Then,two-dimensional gel electrophoresis（2-DE） was used to identify the differentially expressed proteins between the wild-type strain and the mutant strain.Results The null-mutant strain of gadB was successfully constructed that could not use glycerol as the only carbon source.Sereny test revealed that the virulence of gadB mutant was slightly weaker than that of the wild-type strain.A total of 10 proteins were identified as differentially expressed proteins according to the comparative proteomic results.Conclusion Deletion of gadB can influence the expression of some proteins.These data provide some clues to the function research of gadB. |
| Keywords: | Shigella flexneri GadB λ-Red recombinant system knockout two-dimensional gel electrophoresis |
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