【摘要】 为制备双峰驼Toll样受体3(TLR3)的抗体，分析其在肾脏中的表达特征。用生物信息学分析及原核表达方式，分析、表达和纯化了双峰驼TLR3重组蛋白，并制备多克隆抗体，用间接酶联免疫吸附试验和免疫印迹法测定抗体效价及其特异性，利用制备的多克隆抗体采用免疫组织化学染色观察TLR3在双峰驼肾脏组织中的表达。结果显示，得到的目的蛋白大小与预测结果相符，为74 ku, IPTG最佳诱导浓度为0.2 mmol/L,最佳诱导时间为6 h,重组蛋白主要以包涵体形式表达，抗体效价达1∶64 000;免疫印迹结果显示，原核表达的TLR3蛋白和双峰驼肾脏组织中表达的TLR3蛋白均能被该抗体特异性识别，TLR3在双峰驼肾小管上皮细胞和肾小球系膜细胞均有表达。研究结果为进一步探究双峰驼TLR3在肾脏中发挥作用的机制积累了资料。更多还原

【Abstract】 In order to prepare an antibody against Bactrian camel Toll-like receptors 3(TLR3),its expression characteristics in the kidney was analyzed.In this study, bioinformatics analysis and prokaryotic expression were used to analyze, express and purify the Bactrian camel TLR3 recombinant protein, and the polyclonal antibody was prepared.The antibody titer and specificity were detecteded by indirect ELISA and Western blot.The expression of TLR3 in the kidney of Bactrian camel was observed by immunohistochemical staining.The results showed that the size of the obtained target protein was the same as expected size 74 ku, the optimal induction concentration of PTG was 0.2 mmol/L and the induction time was 6 h.The recombinant protein was mainly expressed in the form of inclusion body, and the titer of antibody was 1∶64 000.Western blot results showed that both prokaryotic TLR3 protein and TLR3 protein expressed in Bactrian camel kidney tissue could be specifically recognized by this antibody.TLR3 was expressed in renal tubular epithelial cells and glomerular mesangial cells of Bactrian camel.The results accumulated data for further study on the mechanism of TLR3 in the kidney of Bactrian camel.更多还原