滑液支原体WVU1853株热不稳定延伸因子的表达及黏附特性分析

doi:10.11843/j.issn.0366-6964.2022.02.018

Abstract

Abstract: The aim of this study was to investigate the adhesive function of elongation factor thermo unstable (EF-Tu) of Mycoplasma synoviae (MS). The primer pairs Tu-F/Tu-R were designed according to the sequence of EF-Tu gene of MS WVU1853 strain in GenBank. The MS EF-Tu gene was amplified by PCR and cloned into pET-28a (+). The recombinant plasmid pET-EF-Tu was constructed and transformed into Escherichia coli (E. coli) BL21(DE3). Then the recombinant proteins (rMSEF-Tu) were expressed after induction by IPTG, and the expression product was purified and used to immunize New Zealand rabbits to prepare anti-serum. Subsequently, the immunogenicity of the rMS EF-Tu and the distribution of EF-Tu in MS were respectively analyzed by Western blot, ELISA and immunofluorescence tests. The complement mediated mycoplasmacidal activity of recombinant protein antiserum was assessed by complement mediated bactericidal assay, and the adhesion function of MS EF-Tu was evaluated by adhesion and inhibition assay. The results showed that MS EF-Tu was expressed mainly in soluble form in E. coli. The relative molecular mass of rMS EF-Tu was about 43 ku, and had good immunogenicity. The anti-rMS EF-Tu serum has complement-mediated mycoplasmacidal activity. In addition, the MS EF-Tu was distributed both in MS membrane and cytoplasm, and was an adhesion-related protein of MS. This study showed that EF-Tu was an immunogenic protein related to MS membrane surface adhesion. The verification of its biological function laid a foundation for further study of MS EF-Tu.

Key words: Mycoplasma synoviae, elongation factor thermo unstable (EF-Tu), immunogenicity, membrane protein, adhesion

CLC Number:

S852.62

YUE Yahui, XING Xiaoyong, WU Xiaochun, WEN Fengqin, ZHANG Hongyan, LONG Cuiqin, ZHANG Li, MA Haiyun, BAO Shijun. Expression of Elongation Factor Thermo Unstable of Mycoplasma synoviae WVU1853 Strain and Analysis of Its Adherence Characterization[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 520-528.

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Expression of Elongation Factor Thermo Unstable of Mycoplasma synoviae WVU1853 Strain and Analysis of Its Adherence Characterization

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