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牛坏死杆菌43K OMP的截短表达及多克隆抗体的制备

Expression and Polyclonal Antibody Preparation of Truncated 43K OMP Protein from Fusobacterium necrophrum

  • 摘要: 为制备牛坏死杆菌截短43K OMP多克隆抗体,以A25菌株基因组DNA为模板,特异性扩增43K OMP基因的4个相互重叠的截短片段,构建原核表达载体,利用大肠杆菌原核表达系统对截短基因进行表达并纯化蛋白,同时免疫家兔制备多克隆抗体。结果表明:43K OMP基因截短片段经IPTG诱导成功表达,蛋白大小分别为32 kDa、30 kDa、28 kDa和30 kDa,运用蛋白制备的多克隆抗体43K OMP-1、43K OMP-2、43K OMP-3和43K OMP-4的效价分别为1∶25 600、1∶51 200、1∶25 600和1∶51 200,且制备的抗体能识别天然43K OMP。

     

    Abstract: In order to prepare the polyclonal antibody of Fusobacterium necrophrum,so as to promote the research of genetic engineering vaccine from Fusobacterium necrophrum,the study used A25 gene DNA as template to amplify four fragments of 43 K OMP gene overlap each other, and constructed prokaryotic expression vector.The purified protein was immunized to rabbits by prokaryotic expression system of E.coli,and then the polyclonal antibody was prepared and identified.The results showed that the truncated 43 K OMP proteins were successfully expressed, and the length of the truncated protein size was 32 kDa, 30 kDa, 28 kDa and 30 kDa respectively.The polyclonal antibodies 43 K OMP-1,43 K OMP-2,43 K OMP-3 and 43 K OMP-4 prepared by the protein were 1∶25 600,1∶51 200,1∶25 600,1∶51 200.The prepared antibody can recognize natural 43 K OMP,which lays a foundation for the research of genetic engineering vaccine of Fusobacterium necrophrum.

     

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