摘要
牛支原体(M.bovis)在感染和致病过程中其粘附蛋白起到关键的作用。为鉴定M.bovis膜表面粘附蛋白,本研究利用生物学软件对M.bovis全基因序列进行分析,预测其中一个891 bp的开放阅读框架编码一个约33 ku的具有粘附特征的假定蛋白,并将其命名为P33蛋白。我们通过PCR扩增该基因并进行原核表达,获得了重组目的蛋白(rP33)。Western blot鉴定结果显示,P33蛋白定位于M.bovis菌体表面;激光共聚焦检测表明,rP33具有吸附胎牛肺细胞(EBL)的能力,其吸附作用可以被抗rP33血清特异性抑制。而且,以rP33为包被抗原的ELISA对EBL细胞膜成份的检测能够被抗rP33血清抑制,并呈抗体浓度依赖关系。本研究结果表明P33是一个新发现的具有粘附作用的M.bovis膜表面相关蛋白。
The surface protein of M.bovis played an important role in the entry and infection to the host cells. In this study, we identified an 891 bp open reading frame encoding a 33 ku protein （P33）. The p33 gene was cloned and expressed in E.coli to express recombinant P33 （rP33）. Western blot analysis using a rabbit antibody against rP33 demonstrated that P33 is a membrane protein. In addition, immunostaining visualized via confocal laser scanning microscropy showed that rP33 was able to adhere to embryonic bovine cells （EBL） and this was also confirmed by a sandwich ELISA. In summary, a new surface-localized adhesion protein of M. bovis was identified which would facilitate future study on the pathogenic mechanism of M. bovis.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2014年第9期679-683,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
国家自然科学基金(31072131)
关键词
牛支原体
原核表达
粘附
Mycoplasma bovis
prokaryotic expression
adhesion
