【摘要】：目的:尋求一種核受體家族蛋白NR4A1 DNA結(jié)合域(NR4A1 DNA binding domain,NR4A1-DBD)表達(dá)純化的方法。方法:構(gòu)建融合蛋白PET28a-NR4A1-DBD表達(dá)載體,分別通過鎳親和層析、陽離子交換層析及凝膠過濾的方法對(duì)目的蛋白進(jìn)行純化。結(jié)果:PET28a-NR4A1-DBD蛋白在低溫誘導(dǎo)(24℃)時(shí)多以可溶性形式存在,通過此純化,可達(dá)到每升培養(yǎng)基產(chǎn)生2~3 mg蛋白,純度高達(dá)95%以上。結(jié)論:鎳親和層析是一種可靠、實(shí)用的蛋白表達(dá)純化方法,后續(xù)使用陽離子交換層析及凝膠過濾可以進(jìn)一步提高蛋白純度。
[Abstract]:Aim: to explore a method for the expression and purification of nuclear receptor family protein NR4A1 DNA binding domain (NR4A1 DNA binding domain NR4A1-DBD). Methods: the fusion protein PET28a-NR4A1-DBD expression vector was constructed and purified by nickel affinity chromatography, cation exchange chromatography and gel filtration. Results the protein of 1: PET28a-NR4A1-DBD existed in soluble form when induced at low temperature (24 鈩，

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