title: [Expression and purification of FOXO1 DNA binding domain and its DNA properties].
abstract: To express and purify FOXO1 DNA binding domain (DBD) and to evaluate its DNA binding properties.  Methods: Gene sequence of FOXO1-DBD was optimized and the recombinant strains were induced at low temperature to obtain soluble protein of FOXO1-DBD. Then, FOXO1-DBD protein was purified through Ni affinity chromatography and cation exchange. Finally, the DNA binding properties of FOXO1-DBD was evaluated by electrophoretic mobility shift assay (EMSA).  Results: Most of soluble proteins were obtained by optimizing its genes sequence and induction at 21 ℃. Over 95% purity of FOXO1-DBD protein was obtained through two-steps purification. Purified protein exhibited good binding property to the DNA binding motif (G/ATAAACA) of FOX family.  Conclusion: An efficient expression and purification method for FOXO1-DBD is established, and the complexity of FOXO1 protein in recognition of DNA is verified.
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Yinuer
authors.initials: Y
authors.lastname: Ha
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Yongheng
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Zhuchu
authors.initials: Z
authors.lastname: Chen
mesh_terms: D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins; D024202:Electrophoretic Mobility Shift Assay; D000071161:Forkhead Box Protein O1 / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D000072417:Protein Domains; D011994:Recombinant Proteins
chemical_list: D004268:DNA-Binding Proteins; D000071161:Forkhead Box Protein O1; D011994:Recombinant Proteins; D004247:DNA