摘要
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28kD,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∶62500。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。
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 trans- formed into BL21. After 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 control. These suggest that immunoneutralization of myostatin during embryonic development could improve the growth performance of large mouth bass.
出处
《生物技术通报》
CAS
CSCD
北大核心
2009年第12期129-133,共5页
Biotechnology Bulletin
基金
国家科技支撑项目(2006BAD01A1209)
国家科技基础条件平台工作项目(2005DKA21103)
广东省自然科学基金项目(7301732)
中国水产科学研究院水产种质资源与养殖技术重点开放实验室开放基金项目
关键词
大口黑鲈
MSTN
原核表达
多克隆抗体
仔鱼
生长
Large mouth bass Myostatin Prokaryotie expression Polyelonal anti-myostatin antibody Fry Growth
