猪鼻支原体表面可变脂蛋白vlpA黏附宿主细胞功能研究

doi:10.11843/j.issn.0366-6964.2016.09.019

Abstract

Abstract:

Mycoplasma hyorhinis (M.hyorhinis) is a swine pathogen that can cause multiple chronic inflammation including polyserositis，pneumonia，arthritis and otitis media.It is also a potential pathogen for human beings，associated with various human cancers.In previous study，vlp family has been revealed to play a role in mediating cytoadhesion of M.hyorhinis.Herein，we furtherly studied the cytoadhesion function of vlpA，one of the vlp family member，particularly how the repeat time of repetitive unit impact the function.According to the sequence published on GenBank，the vlpA gene was amplified from bacterial genome.Furthermore，vlpA genes containing 0，3，6，9 and 12 times of repetitive unit in region Ⅲ were artificially synthesized.The genes were inserted into pET-32a(+) and expressed in Escherichia coli.At the same time，a peptide containing 2 times of repetitive unit in region Ⅲ was artificially synthesized.All the recombinant proteins，as well as the peptide were detected for their adherence to PK-15 cell by using the indirect immunofluorescence assay and microtiter plate adherence assay.All the recombinant proteins have been successfully induced for expression and purified by affinity chromatography.Adherence of vlpA to PK-15 cell was observed by the indirect immunofluorescence assay.vlpA0 could bind PK-15 cell in the microtiter plate adherence assay，but not the peptide containing 2 times of repetitive unit in region Ⅲ.Furthermore，we detected the adherence of recombinant proteins vlpA3，vlpA6，vlpA9，vlpA12 containing 3，6，9 and 12 times of repetitive unit in region Ⅲ by using the microtiter plate adherence assay，the results showed that all of the four recombinant proteins could adhere to the cell，but the adherence was lower than the recombinant protein vlpA0.Additionally，as the repeat times increased，the adherence significantly decreased.The results of this study suggest that vlpA is one of the adhesion factors of M.hyorhinis.The region Ⅱof VlpA contains cytoadhesion site.The adhesion ability of the whole molecule significantly decreased as the times of repetitive unit in region Ⅲ increased.The results of this study laid the foundation for further study on the pathogenic mechanism of M.hyorhinis.

Key words: M.hyorhinis, variable lipoprotein vlpA, cytoadhesion, the number of repeat times

CLC Number:

S852.62

ZHANG Bi-xiong，XIONG Qi-yan，WANG Jia，JI Yan，NI Bo，WEI Yan-na，FENG Zhi-xin，LIU Mao-jun，SHAO Guo-qing. The Function of the Variable Lipoprotein A of Mycoplasma hyorhinis in Adherence to Host Cell[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(9): 1897-1904.

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The Function of the Variable Lipoprotein A of Mycoplasma hyorhinis in Adherence to Host Cell

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