摘要
为了研究绵羊Follistatin基因的功能及与MSTN的结合能力,克隆了FS结构域1及结构域2,构建了405 bp插入序列的原核表达重组质粒pET-FS N+D1,以及630 bp插入序列的pET-FSN+D1+D2。通过SDS-PAGE及Western blot检验,在IPTG诱导表达。结果显示:分别获得了带有6×His的45 kDa和51 kDa融合蛋白,融合蛋白主要以包涵体形式存在。经His亲和层析分别纯化了pET-FSN+D1,pET-FSN+D1+D2,目的蛋白含量分别达2.58μg/mL和5.98μg/mL,为进一步研究Follistatin基因的功能及与MSTN的结合能力提供了基础。
Summary：In order to study the structure and function of Follistatin in sheep,a fragment of about 405 bp consisted of the coding sequences of N-terminal peptide and domain 1, as well as another of 630 bp long consisted of the coding sequences of N- terminal peptide , domain 1 and domain 2 of the target protein were subcloned into the prokaryotic expression vector pET41a, named pET-FSN＋D1 and pET-FSN＋D 1 ＋D2, respectively. The SDS-PAGE and Western blot analysis identified the 45 kDa and 51 kDa fusion protein of different follistatin domains, and the recombinant protein were mainly occurred in the form of inclusion body. After the renaturation and purification employing the His afinity chromatography purification system, two protein solutions contanning the target protein at a level of 2.58 Ixg/mL and 5.98 Ixg/mL respectively were harvested. The study provide useful basis for the further insight into the function of Follistatin as well as the combining capacity of the different domains with MSTN in the future~
出处
《中国草食动物科学》
CAS
2012年第4期5-8,共4页
China Herbivore Science
基金
新疆维吾尔自治区青年基金项目(No:2010211B31)
作者简介
阿米乃·亚日（1986-），女，硕士研究生。
通讯作者：张宁，女，助理研究员。Email：xjzhangning@126．com
