【摘要】 利用PCR方法扩增乳源致病性金黄色葡萄球菌nEBPS全基因,将其定向克隆至原核表达载体pET30a(+)中,鉴定正确后,转化BL21表达菌,经IPTG诱导获得了以可溶性表达的重组蛋白。通过Ni NTA Purification System纯化重组蛋白,纯化蛋白质量浓度为2.16g/L。纯化蛋白经免疫印迹检测显示重组蛋白能够被牛源金黄色葡萄球菌阳性血清识别,具有良好的反应性。将纯化的重组蛋白免疫新西兰白兔制备多克隆抗体,间接ELISA测定抗体效价为1∶25 600,凝集试验测定抗体效价为1∶128。更多还原

【Abstract】 To expression of N-terminal elastin binding proteins of Staphylococcus aureus(nEBPS)and prepare polyclonal antibody,the nEBPS was amplified by PCR and directionally cloned into the pET30avector.After characterization,the recombinant plasmid was transformed into E.coli BL21(DE3),the recombinant nEBPS protein was expressed in soluble body form after induction with IPTG.The concentration of the purified recombinant protein was 2.16g/L.Western blotting showed that purified protein could react specifically to Staphylococcus aureus positive serum.The polyclonal antibodies to the nEBPS protein was prepared by inoculating rabbits with purified recombinant protein.The polyclonal antibodies was generated with antibody titres of 1∶25 600in ELISA and 1∶128in agglutination test.The recombinant protein and the polyclonal antibody obtained in this study could be used for detection Staphylococcus aureus,and could be used to further study the structure,function and epitope mapping of nEBPS.更多还原