NR4A1 DNA结合域的表达纯化
发布时间：2018-07-25 20:51
【摘要】：目的:寻求一种核受体家族蛋白NR4A1 DNA结合域(NR4A1 DNA binding domain,NR4A1-DBD)表达纯化的方法。方法:构建融合蛋白PET28a-NR4A1-DBD表达载体,分别通过镍亲和层析、阳离子交换层析及凝胶过滤的方法对目的蛋白进行纯化。结果:PET28a-NR4A1-DBD蛋白在低温诱导(24℃)时多以可溶性形式存在,通过此纯化,可达到每升培养基产生2~3 mg蛋白,纯度高达95%以上。结论:镍亲和层析是一种可靠、实用的蛋白表达纯化方法,后续使用阳离子交换层析及凝胶过滤可以进一步提高蛋白纯度。
[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，
本文编号：2145050
[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，
本文编号：2145050
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