铜绿假单胞菌潜在Ⅵ型分泌系统效应蛋白PA0423的鉴定及功能研究  

Identification and Functional Study of T6SS Effector Protein PA0423 of Pseudomonas aeruginosa

作　　者：王彩虹 蒋梦媛 邵煜涵 WANG Cai-hong;JIANG Meng-yuan;SHAO Yu-han(Beijing Friendship Hospital,Capital Medical University,Beijing 100050)

机构地区：[1]首都医科大学附属北京友谊医院热带医学研究所,北京100050

出　　处：《生物技术通报》2024年第4期297-305,共9页Biotechnology Bulletin

基　　金：北京市医管中心青苗项目(QML20230105)。

摘　　要：【目的】探讨Ⅵ型分泌系统(type Ⅵ secretion system, T6SS)新的效应蛋白及其功能。【方法】以铜绿假单胞菌PAO1为研究对象,通过基因敲除、免疫印迹、免疫共沉淀、种内和种间竞争、大肠杆菌毒性实验以及结晶紫生物膜等实验探索PA0423的分泌方式和功能。【结果】免疫印迹和免疫共沉淀实验表明PA0423可以与PA0262相互作用,且PA0423的分泌依赖于H2-T6SS;竞争实验表明PA0423是H2-T6SS依赖的抗菌效应蛋白,且具有种间竞争优势;PA0423具有大肠杆菌毒性且PA0422为其免疫蛋白;PA0423能够影响细菌生物膜的形成。【结论】PA0423为H2-T6SS依赖的抗菌效应蛋白,其具有杀菌功能且影响细菌生物膜的形成。【Objective】The objective of this study is to investigate the novel effector proteins and their functions in the type VI secretion system(T6SS).【Method】The secretion mode and function of PA0423 were explored by gene knockout,immunoblotting,coimmunoprecipitation,intraspecific and interspecific competition,Escherichia coli toxicity assays and crystal violet biofilm assays.【Result】Western blotting and co-immunoprecipitation experiments showed that PA0423 interacted with PA0262,and the secretion of PA0423 depended on H2-T6SS.PA0423 is a H2-T6SS-dependent antimicrobial effector protein with interspecific competitive advantages.PA0423 is toxic to Escherichia coli and PA0422 is its immune protein.PA0423 affected the formation of bacterial biofilms.【Conclusion】PA0423 is a H2-T6SSdependent antimicrobial effector protein,which has bactericidal function and affects the formation of bacterial biofilms.

关 键 词：铜绿假单胞菌 Ⅵ型分泌系统 PA0423 效应蛋白 

分 类 号：R378[医药卫生—病原生物学] Q936[医药卫生—基础医学]