中国畜牧兽医2017，Vol.44Issue(2)：371-376,6.DOI:10.16431/j.cnki.1671-7236.2017.02.010
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
Expression of N-terminal Domain and C-terminal Domain of Mycoplasma ovipneumoniae p60 Protein and Initial Identification on Their Immunologic Activity
摘要
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 SDSPAGE,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.关键词
绵羊肺炎支原体/p60蛋白/表达Key words
Mycoplasma ovipneumoniae/p60 protein/expression分类
农业科技引用本文复制引用
麻昌姣,郑佳琪,黄海碧,王晓晖,白帆,郝永清..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[J].中国畜牧兽医,2017,44(2):371-376,6.基金项目
内蒙古自治区科技计划“舍饲肉羊疫病动态防控关键技术研究”(201502070) （201502070）
国家科技支撑计划“重点牧区”生产生态生活“保障技术集成与示范”(2012BAD13B00) （2012BAD13B00）
