摘要
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： To investigate the function of mP24, a specific rabbit polycional antibody of mP24 was obtained from a prokaryotic expression construct of mouse mP24. Methods： mP24, a novel mouse gene, was amplified by PCR and inserted into an inducible prokaryotic expressive plasmid pET-28a （＋）. The recombinant plasmid was transformed into the E.coli BL21 （DE3）. mP24-HIS fusion protein was induced by IPTG, purified and renatured by Ni＋ affinity column. The purified mP24 protein was used to immunize New Zealand rabbits for preparing polyclonal antibody. The specificity and potency of polyclonal antibody was evaluated by Western blot and ELISA. Results： The mP24 fusion protein was inducible expressed and purified. The polyclonal antibody against mP24 was obtained successfully with the titer 〉1：9000, the antibody is specific against mp24 with Western blot. Conclusions： A specific polyelonal antibody was obtained; it will be valuable for the further study on the biological function of mP24.
出处
《现代生物医学进展》
CAS
2009年第13期2428-2431,共4页
Progress in Modern Biomedicine
基金
哈尔滨工业大学校创新项目(GFQQ18400017
2008年)
