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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。
Objective To investigate the potential function of gadB gene and its effect on the pathogenicity of Shigella flexneri 2a 301. Methods The gadB deletion mutant of Shigella flexneri 2a 301 was constructed by using λ-Red recombination system. Subsequently, the basic phenotypic test was carried out to evaluate its virulence. Finally, mutants and wild-type strains were prepared. Two-dimensional electrophoresis was used to compare the differences in protein expression profiles. Results The gadB gene deletion was successfully obtained. The biochemical experiments showed that the mutant strain could not utilize glycerol fermentation. The guinea pig cornea experiment showed that the mutant strain had a slightly lighter inflammatory response than the wild strain. Ten differential proteins were found in the whole-cell protein expression profile. Conclusion Two-dimensional electrophoresis was used to identify some proteins that may be related to GadB, providing clues for the potential function of gadB.