奶牛边缘无浆体MSP2的原核表达及抗原性鉴定

Expression and immunogenicity analysis of membrane surface protein 2 of Anaplasma marginale

为克隆和原核表达牛边缘无浆体(A.marginale)膜表面蛋白2(MSP2),并分析其免疫原性,本研究通过PCR方法扩增A.marginale的MSP2基因,将其连接到pGEX 6p-1载体中,转化感受态细胞BL21(DE3),经IPTG诱导表达,进行SDS-PAGE分析,将纯化的MSP2重组蛋白免疫小鼠。结果表明表达并纯化了A.marginaleMSP2,其免疫原性良好,为制备单克隆抗体及亚单位疫苗的研制奠定了基础。

To express the membrane surface protein 2 （MSP2） of Anaplasma marginale and identify its immunogenicity. The MSP2 gene of A.marginale was amplified by PCR and cloned into pGEX-6P-1 vector for expression in E.coli. SDS-PAGE analysis showed that the recombinant protein was about 68 ku, which reacted positively with anti- A.marginale serum detected by western blot. The expression of A.marginale MSP2 provided a basis for monoclonal antibody and subunit vaccine development against A.marginale.