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目的:本研究是为后续实验中深入研究小鼠mP24基因的功能及活性提供mP24多克隆抗体。方法:mP24是小鼠中新发现的新基因,通过构建了pET-28a(+)/mP24原核表达质粒,转化入大肠杆菌BL21(DE3)中,IPTG诱导宿主细菌表达融合蛋白。经过SDS-PAGE分析,结果表明该蛋白能以可溶性蛋白形式存在。利用镍柱纯化融合蛋白,免疫新西兰兔,制备mP24多抗。结果:Western Blot结果显示,制备的抗小鼠mP24多克隆抗体具有较高的特异性。ELISA实验检测,该多抗对免疫抗原的效价高达1:9000,具有较高的效价。结论:本实验成功制备了mP24的多克隆抗体,为进一步开展mP24基因功能的研究奠定了基础。
OBJECTIVE: The aim of this study is to provide mP24 polyclonal antibody to further study the function and activity of mouse mP24 gene in subsequent experiments. Methods: mP24 was a newly discovered new gene in mice. The prokaryotic expression plasmid pET-28a (+) / mP24 was constructed and transformed into E. coli BL21 (DE3). IPTG induced the expression of the fusion protein in host bacteria. After SDS-PAGE analysis, the results showed that the protein can exist as soluble protein. The fusion protein was purified by nickel column and immunized New Zealand rabbits to prepare mP24 polyclonal antibody. Results: The results of Western Blot showed that the prepared anti-mouse mP24 polyclonal antibody has high specificity. ELISA test, the polyclonal anti-immune titers up to 1: 9000, with high potency. Conclusion: The mP24 polyclonal antibody was successfully prepared in this study, which laid the foundation for the further study of mP24 gene function.