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-FSN+D1,以及630 bp插入序列的pET-FSN+D1+D2.通过SDSPAGE及Western blot检验,在IPTG诱导表达.结果显示:分别获得了带有6xHis的45 kDa和51kDa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2.58 μg/mL和5.98 μg/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.%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+Dl and pET-FSN+Dl+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 μg/mL and 5.98 μg/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 0
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