大口黑鲈肌肉生长抑制素多克隆抗体的制备及其对仔鱼生长影响

肌肉生长抑制素(Myostatin,MSTN)是抑制肌肉生长和发育的生长调控因子.将大口黑鲈(Micropterus salmoides)MSTN成熟肽cDNA改造后克隆到pET32a(+)载体上,构建原核表达质粒,在大肠杆菌BL21中进行表达.SDS-PAGE检测表明重组表达的融合蛋白分子大小约28 kD,含量为菌体总蛋白的34%.用纯化后的表达蛋白免疫新西兰兔制备抗大口黑鲈MSTN多克隆抗体,Western blot和ELISA检测证实所获得的抗体可与抗原蛋白特异性结合,抗体的效价为1: 62 500.将获得的抗体显微注射到大口黑鲈的受精卵卵黄区,结果显示具有促进仔鱼生长的作用.

Production of Polyclonal Anti-myostatin Antibody of Large Mouth Bass and Its Effect on Growth of the Large Mouth Bass Fry

Myostatin,as a negative regulator of myogenesis,inhibits myoblast proliferation and differentiation.In this study,largemouth bass(Micropterus salmoides)myostatin mature peptide cDNA was inserted into the expression vector of pET32a(+),and transformed into BL21After induced by IPTG,the recombinant expression product was detected by SDS-PAGE.The molecule weight of myostatin fusion protein was about 28 kD and accounted for 34% in total protein.Then myostatin protein was purified and injected into rabbit to produce polyclonal anti-myostatin antibody.The antibody showed high affinity to the recombinant myostatin detected by Western blot and ELISA analysis.In order to study the biological activity of myostatin,the myostatin antibody was injected into yolk of one or two cells stage large mouth bass embryo by micro-injection technique.Average weight of the treated fish was significantly heavier than that of the corresponding controlThese suggest that immunoneutralization of myostatin during embryonic development could improve the growth performance of large mouth bass.