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[目的]克隆CbxR基因,进行原核表达,纯化CbxR基因蛋白并制备其多克隆抗体。[方法]CbxR基因克隆至原核表达载体p ET-28a(+),转化大肠杆菌BL21(DE3)诱导表达并纯化,Western blot鉴定分析。制备CbxR蛋白的兔源多克隆抗体,运用间接ELISA方法检测多抗效价,利用Western blot印记检测抗体特异性。[结果]成功获得CbxR基因,在大肠杆菌BL21(DE3)中诱导可大量表达,成功制备CbxR蛋白兔源多克隆抗体,效价为1∶64 000,经Western blot检测表明多克隆抗体特异性良好。[结论]多克隆抗体为CbxR检测及进一步研究CbxR基因功能奠定了基础。
[Objective] The aim of the study was to clone CbxR gene, prokaryotic it, purify CbxR gene protein and prepare its polyclonal antibody. [Method] CbxR gene was cloned into the prokaryotic expression vector p ET-28a (+) and transformed into E. coli BL21 (DE3) for expression and purification. Western blot analysis was used to analyze the expression of CbxR gene. The rabbit polyclonal antibody of CbxR protein was prepared, the multi-titers were detected by indirect ELISA and the antibody specificity was detected by Western blot. [Result] The CbxR gene was successfully obtained and induced in E. coli BL21 (DE3). The polyclonal antibody against CbxR protein was successfully prepared and its titer was 1:64 000. The result of Western blot showed that the polyclonal antibody was highly specific . [Conclusion] The polyclonal antibodies laid the foundation for CbxR detection and further study of CbxR gene function.