CAJ | 학술논문

为进一步研究延长因子－1（elongation factor －1α，EF －1α）在调控心脏发育和果蝇心脏功能中发挥的作用，制备延长因子－1的抗体.利用生物信息学选择果蝇 EF －1α基因抗原亲水区，并设计出特异性引物，PCR 扩增出的片段克隆到原核表达 pET －28a 载体中，转入 E ．coli 中后通过 IPTG 诱导融合蛋白表达，将 His － EF －1α融合蛋白免疫新西兰大白兔制备多克隆抗体，再用 Western Blot 检测抗体的效价和特异性.研究结果表明，获得了 EF －1α原核表达重组融合蛋白以及高效价的、特异性兔抗 EF －1α多克隆抗体. EF －1α多克隆抗体的效价可以达到1比1500，并且有很强的特异性，为后续研究 EF －1α基因在心脏发育和心脏功能中的作用奠定了基础.
위진일보연구연장인자－1（elongation factor －1α，EF －1α）재조공심장발육화과승심장공능중발휘적작용，제비연장인자－1적항체.이용생물신식학선택과승 EF －1α기인항원친수구，병설계출특이성인물，PCR 확증출적편단극륭도원핵표체 pET －28a 재체중，전입 E ．coli 중후통과 IPTG 유도융합단백표체，장 His － EF －1α융합단백면역신서란대백토제비다극륭항체，재용 Western Blot 검측항체적효개화특이성.연구결과표명，획득료 EF －1α원핵표체중조융합단백이급고효개적、특이성토항 EF －1α다극륭항체. EF －1α다극륭항체적효개가이체도1비1500，병차유흔강적특이성，위후속연구 EF －1α기인재심장발육화심장공능중적작용전정료기출.
In order to study the role that elongation factor -1α (elongation factor - 1 α ,EF - 1 α)played in the regulation of heart development and Drosophila heart function ,the EF - 1α antibody must be prepared .The Drosophila EF - 1 alpha gene antigen hydrophilic region was chose by bioinformatics , and designed specific primers . The PCR fragments were cloned into prokaryotic expression vector ,pET - 28a , then transferred into E .coli .The fusion protein was induced by IPTG (Isopropyl β -D- thiogalactoside) ,which can be used to immunize the New Zealand white rabbits to produce polyclonal antibodies .The titer and specificity of the antibody was analysed by Western blot . It was gained the EF - 1 alpha original expression recombinant fusion protein ,and the high titer and specificity rabbit polyclonal antibody . The titer of EF - 1 alpha polyclonal antibody can be achieved to 1 to 1 500 , still with a strong specificity , which laid the foundation for our further study of its function heart development and cardiac function .