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采用体内表达技术的研究策略筛选了志贺菌福氏2a侵入上皮细胞后诱导表达的基因,用基因突变技术进一步对它们的功能进行了鉴定.结果共筛选到13个胞内诱导基因,其中2个DNA甲基化相关基因、1个代谢相关基因、1个转录调控基因、3个插入成分、3个反义转录体、3个未知功能基因.突变体分析发现,3-甲基糖基化酶u001f,柠檬酸裂合酶的编码基因和未知功能基因wcaJ的突变体在细胞和动物感染实验中都显示其存活增殖能力的降低,表明这些基因可能参与了志贺菌在上皮细胞内的存活和增殖.同时也表明,这种在上皮细胞内的存活增殖能力是志贺菌主要的毒力体现之一.但是,未知功能基因yaiC的突变体只在动物体内表现出明显的毒力缺陷,在上皮细胞内没有明显的增殖缺陷,这表明yaiC基因参与其他毒力作用机制.研究结果对深入认识志贺菌的致病机理有重要意义.
In vivo expression strategy was used to screen the gene induced by Shigella flexneri 2a invaded into epithelial cells and their function was further identified by gene mutation technique.Results Thirteen genes were induced in vitro and in which 2 One DNA methylation related gene, one metabolic related gene, one transcriptional regulatory gene, three inserted components, three antisense transcripts and three unknown functional genes.Mutant analysis showed that 3-methylglycosylation Enzyme u001f, the gene encoding citrate lyase and the mutant wcaJ of unknown function both showed decreased viability in cell and animal infection experiments, indicating that these genes may be involved in the growth of Shigella in epithelial cells Survival and proliferation.It also shows that this survival and proliferation in epithelial cells is one of the main manifestations of Shigella virulence.However, the mutant of unknown function gene yaiC only shows obvious virulence defects in animals , There is no obvious proliferation defects in epithelial cells, indicating that yaiC gene is involved in other virulence mechanisms.The results of this study are of great significance for understanding the pathogenesis of Shigella.