CAJ | 학술논문

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.
기육생장억제소(Myostatin,MSTN)시억제기육생장화발육적생장조공인자.장대구흑로(Micropterus salmoides)MSTN성숙태cDNA개조후극륭도pET32a(+)재체상,구건원핵표체질립,재대장간균BL21중진행표체.SDS-PAGE검측표명중조표체적융합단백분자대소약28 kD,함량위균체총단백적34%.용순화후적표체단백면역신서란토제비항대구흑로MSTN다극륭항체,Western blot화ELISA검측증실소획득적항체가여항원단백특이성결합,항체적효개위1: 62 500.장획득적항체현미주사도대구흑로적수정란란황구,결과현시구유촉진자어생장적작용.
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.