抗果蝇MRJ蛋白单克隆抗体的制备和鉴定

摘要

Preparation of anti-Drosophila MRJ protein monoclonal antibody could be used for research of mrj biological function in Drosophila.The recombinant plasmid pET28a-mrj in Rosetta was induced with IPTG to express MRJ recombinant protein.The recombinant protein purified by Ni-IDA gel column was used to immunized BALB/c mice.Then the spleen cells of immunized mice and mouse myeloma cells SP2/0 were used for cell fusion.A hybridoma cell line which secrete anti-mrj monoclonal antibodies was obtained through subcloning and screening progress.The monoclonal antibodies was identified by Elisa and Western Blot.The results show that the anti-mrj monoclonal antibodies can binding to MRJ protein of prokaryotic and eukaryotic cells specificity.It could be used for biology function research of mrj gene.%制备抗果蝇MRJ蛋白单克隆抗体可用于研究果蝇mrj的生物学功能,使用IPTG诱导重组质粒pET28a-mrj在大肠杆菌Rosetta中表达,重组蛋白经过Ni-IDA凝胶柱亲和纯化后免疫BALB/c小鼠.然后取免疫好的小鼠的脾脏细胞与小鼠骨髓瘤细胞SP2/0融合,经克隆和筛选获得了能分泌抗果蝇MRJ蛋白单克隆抗体的杂交瘤细胞株.腹水制备后获得单克隆抗体,通过ELISA和Western Blot对所获得的抗体进行鉴定,结果表明所制备单克隆抗体能够特异性结合于原核及真核细胞表达的MRJ蛋白,可用于研究mrj基因的生物学功能.