重组猪鼻支原体P37蛋白的原核表达及免疫原性分析  

Prokaryotic expression and immunogenicity identification of Mycoplasma hyorhinis recombinant P37 protein

作　　者：熊祺琰[1] 刘占军[1] 马庆红[1] 冯志新[1] 刘茂军[1] 白方方[1] 邵国青[1] 

机构地区：[1]江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室/国家兽用生物制品工程技术研究中心,江苏南京210014

出　　处：《中国预防兽医学报》2013年第10期840-844,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：江苏省自然科学基金(BK20130702);江苏省农业科技自主创新资金[cx(12)1001-05]

摘　　要：P37是猪鼻支原体(Mhr)的主要免疫原之一,也是Mhr中目前唯一已知的与肿瘤直接相关的抗原。为原核表达Mhr P37蛋白并分析其免疫原性,本研究通过人工合成能够在E.coli中优势表达P37蛋白的编码序列,以pET-28a(+)为载体对P37重组蛋白进行原核表达。SDS-PAGE及western blot鉴定结果表明,表达的P37重组蛋白约为45.97 ku并能够被Mhr高免血清识别。利用纯化的重组蛋白为包被抗原初步建立间接ELISA检测方法,检测结果显示与Mhr高免血清呈强阳性反应。此外,将P37重组蛋白免疫小鼠,能够诱导高滴度特异性抗体,表明该蛋白具有良好的免疫原性。本研究结果为Mhr检测方法的建立、P37蛋白功能研究以及P37相关重组肿瘤疫苗和Mhr疫苗的研制奠定基础。Mycoplasma hyorhinis is the pathogen to induce multiple inflammatory diseases inculding polyserositis and arthritis in pigs, and has also been proved to be highly related to several kinds of human cancers. P37 is a major antigen of M.hyorhinis, which is the only identified tumor-associated protein in M.hyorhinis yet. In the present study, a gene fi＇agment encoding P37 was synthesized and inserted into the pET-28a（＋） for expression in E.coli. SDS-PAGE and western blot assays showed that the recombinant P37 was 45.97 ku and reacted with M.hyorhinis positive serum. An indirect-ELISA method using the purified P37 as coating antigen was established and the detection result of which demonstrated a strong positive reaction against M.hyorhinis positive serum. Furthermore, a high level of antibody was induced in mice immunized with the recombinant P37 protein. These results would facilitate future study of P37 and development of subunit vaccine against M.hyorhinis infection.

关 键 词：猪鼻支原体 P37蛋白 原核表达 免疫 

分 类 号：S852.62[农业科学—基础兽医学]