Initial Identification of Adhesion-related Proteins of Mycoplasma bovis of Guizhou Strains
Mycoplasma bovis(M.bovis)is an important pathogen that causes a variety of bovine diseases including bovine mastitis and calf pneumonia.Adhesion proteins are often thought to play key roles in the pathogenic mechanisms of M.bovis.Therefore,this study aimed to investi-gate four putative proteins of M.bovis Guizhou strain(GZ-2),namely M27,M32,M498,and M663,through prokaryotic expression in Escherichia coli BL21(DE3)and purification,subcel-lular localization and adhesion studies.The results showed that recombinant M27,M32,M498 and M663 proteins were successfully expressed and purified;M27,M32 and M498 proteins all showed specific blotting bands in total proteins,cytosolic and cytoplasmic proteins of M.bovis,whereas M663 was undetectable;Immunostaining results showed that M27,M32,M498 and M663 proteins as well as M.bovis adhered to embryonic bovine lung(EBL)cells under confocal laser scanning microscopy,and the rabbit anti-M27,M32,M498 and M663 antibodies were able to inhibit the adherence of these proteins and M.bovis to the EBL cells.This was further con-firmed by flow cytometry analysis with identical results.In conclusion,M27,M32 and M498 proteins are adhesion-associated proteins of M.bovis;of note,although M663 proteins were not detected in the Western blot analysis,it still is able to adhere to EBL cells and simultaneously in-hibits the adhesion of M.bovis to EBL cells.This finding provides a basis of reference for the se-lection of target proteins for the diagnosis of M.bovis-associated diseases and the development of preventive vaccines.
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