小鼠nulp1基因多克隆抗血清的制备及检测

Preparation and Detection of Polyclonal Antiserum of Nulp1 Gene in Mouse

为了在小鼠模型中更深入地研究nulp1蛋白在心脏发育中的功能,采用PCR技术扩增出小鼠nulp1基因的部分编码区并连接入pET-28a表达载体中,之后将重组质粒转入大肠杆菌(E.coli)通过IPTG诱导表达Hisnulp1融合蛋白,对该融合蛋白采用Ni-IDA凝胶柱层析纯化后,免疫新西兰兔制备了多克隆抗体,并用western blotting对抗体进行分析.结果表明,获得了高效价的特异性兔抗nulp1多克隆抗体.这为nulp1功能的进一步研究奠定了基础.

To better understand the function of the nulp1 protein in heart development by using Mouse as a model , here , a fraction of the encoded region in the Mouse nulp1 was obtained by PCR amplification , then the fragment was inserted into pET -28a vector to establish the expressing system . The recombinant plasmid was transformed into E . coli and fusion protein was induced by IPTG . The purified protein was obtained by treating New Zealand white rabbits to prepare antibody . The antibody was assayed by Western blotting . The result show that the polyclonal antibody is of high sensitivity and specificity , which lays a solid foundation for the further studies of nulp1 function .