【摘要】 利用BL21/pET-32a-Nanog表达菌株诱导表达Nanog融合蛋白,经破碎、高速离心、洗涤,获得Nanog蛋白粗制品,用8 moL/L尿素变性溶解Nanog融合蛋白,离心取上清,进行Histrap HP亲和层析纯化,复性,经SDS-PAGE和Western Blot鉴定得到了纯度为97%以上的重组小鼠Nanog蛋白。建立了一种可行的从包涵体中纯化重组小鼠Nanog蛋白的方法,为Nanog蛋白的活性检测及其基因表达调控机理的功能研究奠定基础。更多还原

【Abstract】 In this experiment,Nanog fusion protein was expressed by BL21/pET-32a-Nanog,and the crude Nanog protein was obtained from inclusion body by crushing,high speed centrifuging and washing.Then the inclusion body was dissolved by 8M urea and centrifuging.The supernatant,which be used to Histrap HP column gradual dialysis,renatured and concentrated,was got by centrifuging.the purity of Nanog fusion protein was analyzed by SDS-PAGE and Western blot,and it is up to 97%.The way of preparing mouse Nanog fusion protein from the inclusion body was established,which was prepared for activation detection and mechanism of gene expression and regulation.更多还原