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目的表达ｉＮＯＳＮ端片段并制备ｉＮＯＳ特异性抗体。方法将ｉＮＯＳＮ端片段装入原核表达载体ｐＥＴ－２８ａ（＋），在大肠杆菌ＢＬ２１中表达，Ｈｉｓ．ＢｉｎｄＴＭ柱亲和层析分离纯化及凝胶蛋白回收的两步纯化法纯化目的蛋白，使用纯化蛋白制备ｉＮＯＳ多克隆抗体。结果得到具有较高表达量的融合蛋白，经两步纯化获得ｉＮＯＳＮ端融合蛋白纯品，将此蛋白免疫家兔，得到ｉＮＯＳ多克隆抗体，Ｗｅｓｔｅｒｎｂｌｏｔ证实该抗体具有较好的反应性及特异性。结论原核表达ｉＮＯＳＮ端片段制备ｉＮＯＳ抗体具有很好的特异性。
Objective To express iNOSN end-fragments and prepare iNOS-specific antibodies. Methods The iNOSN fragment was inserted into the prokaryotic expression vector pET-28a (+) and expressed in E. coli BL21. Purification of the target protein by BindTM affinity chromatography and gel protein recovery purification using two-step purification method, the purified protein was used to prepare iNOS polyclonal antibody. The result showed that the fusion protein with high expression level was purified by two steps to obtain purified iNOSN fusion protein. Rabbit was immunized with iNOS to obtain the polyclonal antibody against iNOS. Western blot showed that the fusion protein had good reactivity and specificity. Conclusion Prokaryotic expression of iNOSN end-fragment preparation of iNOS antibody has good specificity.