Construction of a bimolecular fluorescence complementation system for Campylobacter jejuni and its application in protein interaction studies
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.
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