大黄鱼肌肉生长抑制素-1前肽原核表达、多克隆抗体制备和功能验证

Large Yellow Croaker MSTN-1 Prodomain Prokaryotic Expression, Polyclonal Antibody Preparation and Antibody Function Identification

肌肉生长抑制素(myostatin,MSTN)在动物肌肉的生长和发育过程中起着重要作用。该研究通过RT-PCR从大黄鱼(Larimichthys crocea)肌肉组织中克隆肌肉生长抑制素-1前肽基因(MSTN-1pros),构建了表达大黄鱼生长抑制素前肽的重组质粒,并在大肠杆菌BL21(DE3)中进行表达,获得了大黄鱼MSTN-1pros融合蛋白。用分离的重组蛋白免疫大白兔,制备了抗大黄鱼MSTN-1pros的多克隆抗体。经蛋白免疫印迹法检测,确定获得的抗体具有较高的特异性。通过对大黄鱼幼鱼的注射实验,发现注射过MSTN-1pros抗体的大黄鱼增重率比对照组降低了20.71%(P<0.05),证明MSTN前肽抗体对大黄鱼的生长具有抑制作用。实验结果表明,MSTN前肽蛋白可以用于养殖鱼类的生长调控。

Myostatin（MSTN） plays an important role in the muscle growth and development of animals. MSTN-1 prodomain（MSTN-1pros） gene was cloned from the muscle tissue of large yellow croaker（Larimichthys crocea） by RT-PCR. Recombinant expression vectors containing large yellow croaker MSTN-1pros were constructed and transformed into E. coli BL21（DE3）. The recombinant fusion protein was induced with IPTG and isolated by SDS-PAGE. The isolated fusion protein was used to raise the polyclonal antibody of MSTN-1pros by immunizing rabbit. The result of Western blot showed that the polyclonal antibody was highly specific. The weight gain rate of the juvenile large yellow croaker injected with MSTN-1pros antibody was 20.71% lower than that of the control group（P〈0.05）. The results indicated that MSTN-1pros antibody had a biological function of inhibiting the muscle growth of large yellow croaker. It implied that recombinant MSTN-1pros could be employed in aquaculture to improve fish growth.