牛支原体贵州株黏附相关蛋白的初步鉴定

doi:10.11843/j.issn.0366-6964.2024.04.029

Abstract

Abstract: 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 investigate 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, subcellular 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 confirmed 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 inhibits the adhesion of M. bovis to EBL cells. This finding provides a basis of reference for the selection of target proteins for the diagnosis of M. bovis-associated diseases and the development of preventive vaccines.

Key words: Mycoplasma bovis, adhesion protein, prokaryotic expression, screening

CLC Number:

S852.62

LUO Xiaofen, XIE Xiaodong, ZHAO Chao, HU Qian, WANG Yongxuan, RAN Fangfei, HU Pengfei, WEN Ming, ZHU Erpeng, CHENG Zhentao. Initial Identification of Adhesion-related Proteins of Mycoplasma bovis of Guizhou Strains[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1672-1683.

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Initial Identification of Adhesion-related Proteins of Mycoplasma bovis of Guizhou Strains

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