【摘要】 将原核表达质粒pGEX-6P-1-CCL28转化至大肠埃希菌BL21中,经IPTG诱导,获得了GST-CCL28可溶性融合表达产物。利用纯化的GST-CCL28蛋白免疫BALB/c小鼠制备了抗GST-CCL28的多克隆抗体。同时将CCL28基因亚克隆进真核表达载体,经测序验证后命名为pcDNA3.1-CCL28。间接免疫荧光及Western blot试验结果进一步证明,所获得的抗GST-CCL28多克隆抗体能特异性的识别转染pcDNA3.1-CCL28质粒的DF-1细胞内高表达的CCL28蛋白。这一研究为制备抗CCL28单克隆抗体和探究与CCL28相关的生物学功能及其机制奠定了良好的基础。更多还原

【Abstract】 In this study, the prokaryotic expression plasmid pGEX-6 P-1-CCL28 was transformed into BL21 and the soluble fusion expression of GST-CCL28 induced by IPTG was obtained. Moreover, the polyclonal antibody against GST-CCL28 was prepared by immunizing BALB/c mice with the purified GST-CCL28 protein. The chemokine CCL28 gene was also subcloned into eukaryotic expression vector, named as pcDNA3.1-CCL28 after confirmed by sequencing. Indirect immunofluorescence and Western blot assay revealed that the polyclonal antibody against GST-CCL28 generated could specifically recognize the CCL28 protein expressed in DF-1 cells transfected with pcDNA3.1-CCL28. All these data laid a solid foundation for the preparation of monoclonal antibodies against CCL28 and for the exploration of biological functions of CCL28.更多还原