翼螺旋转录因子FOXO1的DNA结合域的表达、纯化及结合特性

发布时间：2018-03-13 18:31

  本文选题：FOXO　切入点：DNA结合域　出处：《中南大学学报(医学版)》2017年01期 　论文类型：期刊论文

【摘要】：目的:探讨翼螺旋转录因子FOXO1的DNA结合域(FOXO1 DNA binding domain,FOXO1-DBD)的表达、纯化及与DNA的结合特性。方法:采用优化FOXO1-DNA的基因序列和低温诱导的方式实现FOXO1-DBD蛋白的可溶性表达,通过镍亲和层析及阳离子交换层析进行纯化,并经凝胶迁移实验(electrophoretic mobility shift assay,EMSA)验证FOXO1-DBD的DNA结合特性。结果:优化后的FOXO1基因在21℃时编码的蛋白大多以可溶性方式表达,通过两步纯化即可获得95%以上纯度的FOXO1-DBD蛋白,纯化的蛋白与含FOX家族DNA结合基序(G/ATAAACA)的DNA序列显示良好的结合特性。结论:建立了FOXO1-DBD蛋白高效表达、纯化的方法,验证了FOXO1蛋白在识别DNA上的复杂性。
[Abstract]:Aim: to investigate the expression, purification and binding characteristics of the DNA binding domain FOXO1 DNA binding domain FOXO1-DBD of pterygoid transcription factor FOXO1. Methods: the soluble expression of FOXO1-DBD protein was achieved by optimizing the gene sequence of FOXO1-DNA and low temperature induction. Nickel affinity chromatography and cationic exchange chromatography were used to purify FOXO1-DBD. The results showed that the DNA binding properties of FOXO1-DBD were verified by gel migration assay. Results: most of the proteins encoded by the optimized FOXO1 gene were expressed in a soluble manner at 21 鈩，

本文编号：1607664