扬州大学学报（农业与生命科学版）2024，Vol.45Issue(4)：1-10,10.DOI:10.16872/j.cnki.1671-4652.2024.04.001
空肠弯曲菌双分子荧光互补系统构建及其在蛋白质互作研究中的应用
Construction of a bimolecular fluorescence complementation system for Campylobacter jejuni and its application in protein interaction studies
摘要
Abstract
In order to study protein interactions in C.jejuni,a bimolecular fluorescence complementation(BiFC)system suit-able for stable and efficient expression in this bacterium was established.By constructing recombinant vectors based on different conditions of gene promoters,fluorescent proteins,and gene complementation,respectively,and introducing them into C.jejuni for the comparative analysis of fluorescence values,we determined the parameters for optimal fluorescence ex-pression,and established the BiFC system with the porA promoter and the plasmid complementation of Venus fluorescent protein.The system was used to explore the interactions of flagellum-expressed proteins of C.jejuni.The results showed that it was known that the fluorescence values of BiFC-flhF-flhG vector,in which flhF interacted with flhG,were signifi-cantly higher than those of the negative control vector for in vivo expression of the bacterium,whereas BiFC-flhF-fliM and BiFC-flhF-fliG did not differ from the negative control,which indicated that the establishment of the BiFC system was successful,suggesting that the interactions of flhF with fliG and fliM did not have mutualistic relationship.In conclusion,the BiFC system for C.jejuni was established in this study,which provides an efficient and convenient method for the protein interactions study of this bacterium.关键词
空肠弯曲菌/BiFC/荧光值/蛋白质互作Key words
Campylobacter jejuni/BiFC/fluorescence values/protein interactions分类
农业科技引用本文复制引用
郭嘉乐,任方哲,李晓飞,郭长彬,焦新安,黄金林..空肠弯曲菌双分子荧光互补系统构建及其在蛋白质互作研究中的应用[J].扬州大学学报（农业与生命科学版）,2024,45(4):1-10,10.基金项目
国家自然科学基金资助项目(32000093、32172939) （32000093、32172939）
扬州市重点研发计划(社会发展)项目(YZ2022059) （社会发展）
扬州大学兽医学学科特区学科交叉课题支持项目(yzuxk202003) （yzuxk202003）
江苏省研究生科研与实践创新计划项目(SJCX21-1632) （SJCX21-1632）
