将扩增的甘蔗花叶病毒P1基因克隆到原核表达载体pET-32a上,转化BL21(DE3),获得重组子pET-32a-P1。超声波结果显示,所得的融合蛋白以可溶性的形式存在。SDS-PAGE检测其表达产物与目的蛋白大小一致,割胶免疫注射家兔得到抗体。间接ELISA测定结果表明稀释51200倍仍呈阳性信号,并通过western blot检测,证明抗体特异性良好。

The target gene was subcloned into the prokaryotic expression vector pET-32a,and transformed into E. coli Bl21(DE3),to obtain recombinant pET-32a-P1.Ultrasound results showed that the fusion protein existed in soluble form.Expression product had the same size with the target protein detected by SDS-PAGE.The antibody against the P1 protein was gained by immunized rabbit with target protein cut from SDS-PAGE.Indirect ELISA measurements proved that the antibody specificity was fine,because of that the dilution of 51200-fold not merely kept positive still,but also passed western blot.