绵羊肺炎支原体p60蛋白N端与C端的表达及其免疫学特性研究

doi:10.16431/j.cnki.1671-7236.2017.02.010

Abstract

Abstract:

In order to analyze the immunological characteristics of lipid-associated membrane protein p60 of Mycoplasma ovipneumoniae, we cloned the N-terminal part and C-terminal part of p60 gene by PCR separately. The overlap-extension PCR was used to mutagenate nucleotides TGA (1 459－1 461 bp) to TGG. Then the PCR product was inserted to the pET-32a(+) plasmid for expression in E.coli BL21(DE3). The purified recombinant protein was analyzed by SDS-PAGE, and the recombinant N-terminal domain of p60 protein was successfully expressed with a mass of molecule of 57 ku, and a molecular of 30.5 ku of the N-terminal domain of p60 protein. Western blotting revealed that the two kinds of proteins could specifically react with positive serum, which confirmed that the recombinant N-terminal domain of p60 protein and the recombinant C-terminal domain of p60 protein both had a good reactionogenicity. This study could provide an effective reference data for screening important immunogenicity protein and establishment available diagnosis methods.

Key words: Mycoplasma ovipneumoniae; p60 protein; expression

CLC Number:

S852.62

MA Chang-jiao, ZHENG Jia-qi, HUANG Hai-bi, WANG Xiao-hui, BAI Fan, HAO Yong-qing. Expression of N-terminal Domain and C-terminal Domain of Mycoplasma ovipneumoniae p60 Protein and Initial Identification on Their Immunologic Activity[J]. , 2017, 44(2): 371-376.

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Expression of N-terminal Domain and C-terminal Domain of Mycoplasma ovipneumoniae p60 Protein and Initial Identification on Their Immunologic Activity

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