马链球菌马亚种FNEB蛋白的截短表达及其免疫保护性研究  

Expression and immunogenicity identification on truncated FNEB protein of Streptococcus equi subsp.equi

作　　者：徐全圆[1] 石明[1] 杨夷平[1] 李娜[1] 刘建华[1] 加尔肯[1] 冉多良[1] 

机构地区：[1]新疆农业大学动物医学学院,新疆乌鲁木齐830052

出　　处：《中国预防兽医学报》2015年第2期132-135,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家星火计划项目(2011GA890008);国家科技支撑计划项目(2012BAD46B00)

摘　　要：为研究马链球菌马亚种FNEB蛋白的免疫保护性,本研究采用PCR方法从马链球菌马亚种新疆分离株中扩增FNEB基因部分片段,克隆于原核表达载体p ET-30a中,转化大肠杆菌BL21(DE3)感受态细胞,以IPTG诱导表达,经SDS-PAGE和western blot分析显示,重组蛋白大小约60 ku,具有良好的抗原性。将纯化的重组蛋白免疫小鼠后,以马链球菌马亚种新疆分离株进行攻毒,结果显示该重组蛋白对免疫组小鼠具有保护力,保护率为65%。本研究克隆了马链球菌马亚种的FNEB截短基因并表达了相应重组蛋白,免疫小鼠后能够提供较好的保护,为重组FNEB蛋白亚单位疫苗的研制奠定基础。For study the immunoprotection of fibronectin-binding （FNEB） against Streptococcus equi subsp, equi in mice, A fragment of FNEB gene was amplified by PCR from S.equi subsp, equi isolated from Xinjiang. The tnmcated FNEB gene was cloned into the pET-30a vector and the resultant recombinant plasmid was transformed into E.coli BL21 （DE3） cells, in which a recombinant FNEB protein （rFNEB） was expressed by inducing with IPTG. SDS-PAGE analysis showed that the expressed rFNEB was about 60 ku which was recognized by the positive serum against S.equi. The mice were immunized intramuscularly three times with purified rFNEB, and challenged with the S.equi subsp, equi. Result showed that the rFNEB provided a 65% protection for immunized mice. The data demonstrated that the FNEB could be used as the antigen for developing the subunit vaccine against S.equi subsp. equi infection in equine.

关 键 词：马链球菌马亚种 FNEB基因 原核表达 

分 类 号：S852.61[农业科学—基础兽医学]