CAJ | 학술논문

为了在斑马鱼模型中更深入地研究Nfatc3蛋白在心脏发育中的功能，采用PCR技术扩增斑马鱼Nfatc3基因的部分编码区并插入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His-Nfatc3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Nfatc3多克隆抗体.这为Nfatc3功能的进一步研究奠定了基础.
위료재반마어모형중경심입지연구Nfatc3단백재심장발육중적공능，채용PCR기술확증반마어Nfatc3기인적부분편마구병삽입pET-28a표체재체중，지후장중조질립전입대장간균(E.coli)통과IPTG유도표체His-Nfatc3융합단백，대해융합단백채용Ni-IDA응효주층석순화후，면역신서란토제비료다극륭항체，병용western blotting대항체진행분석.결과표명획득료고효개적특이성토항Nfatc3다극륭항체.저위Nfatc3공능적진일보연구전정료기출.
To better understand the function of theNfatc3 protein in heart development by using zebrafish as a model, here, a fraction of the encoded region in the zebrafishNfatc3was 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 shows that the polyclonal antibody is of high sensitivity and specificity, which lays a solid foundation for the further studies ofNfatc3function.