發(fā)布時(shí)間：2018-03-13 18:31

  本文選題：FOXO　切入點(diǎn)：DNA結(jié)合域　出處：《中南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年01期 　論文類(lèi)型：期刊論文

【摘要】：目的:探討翼螺旋轉(zhuǎn)錄因子FOXO1的DNA結(jié)合域(FOXO1 DNA binding domain,FOXO1-DBD)的表達(dá)、純化及與DNA的結(jié)合特性。方法:采用優(yōu)化FOXO1-DNA的基因序列和低溫誘導(dǎo)的方式實(shí)現(xiàn)FOXO1-DBD蛋白的可溶性表達(dá),通過(guò)鎳親和層析及陽(yáng)離子交換層析進(jìn)行純化,并經(jīng)凝膠遷移實(shí)驗(yàn)(electrophoretic mobility shift assay,EMSA)驗(yàn)證FOXO1-DBD的DNA結(jié)合特性。結(jié)果:優(yōu)化后的FOXO1基因在21℃時(shí)編碼的蛋白大多以可溶性方式表達(dá),通過(guò)兩步純化即可獲得95%以上純度的FOXO1-DBD蛋白,純化的蛋白與含F(xiàn)OX家族DNA結(jié)合基序(G/ATAAACA)的DNA序列顯示良好的結(jié)合特性。結(jié)論:建立了FOXO1-DBD蛋白高效表達(dá)、純化的方法,驗(yàn)證了FOXO1蛋白在識(shí)別DNA上的復(fù)雜性。
[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，

本文編號(hào)：1607664