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目的初步探讨phoN1基因在志贺菌属中的功能。方法使用λ-Red重组系统构建弗氏志贺菌2a 301株phoN1缺失突变株,通过比较蛋白质组学方法研究突变株与野生株的蛋白表达谱差异,并利用豚鼠角膜炎模型、HeLa细胞竞争侵袭实验评价两者之间的毒力差异。结果成功构建了phoN1缺失突变株,蛋白质学研究发现,phoN1缺失对其他胞内蛋白无显著影响,豚鼠角膜实验和HeLa细胞竞争侵袭实验证实,phoN1缺失不影响志贺菌属的侵袭能力。结论 phoN1在体外生存、侵袭宿主细胞过程中未发挥显著作用,其功能有待进一步研究。
Objective To investigate the function of phoN1 gene in Shigella. Methods The phoN1 deletion mutant of 2a 301 strain of Shigella flexneri was constructed by using λ-Red recombination system. The differences of protein expression between mutants and wild strains were compared by proteomic methods. The competitive invasion of HeLa cells was induced by guinea pig keratitis model Experimental evaluation of the difference between the two virulence. Results The phoN1 deletion mutant was successfully constructed. The results of proteomics showed that phoN1 deletion had no significant effect on other intracellular proteins. The guinea pig cornea assay and HeLa cell invasion assay confirmed that the deletion of phoN1 did not affect the invasive ability of Shigella. Conclusion phoN1 does not play a significant role in the in vitro survival and invasion of host cells, and its function needs further study.