Expression and purification of DNA binding domain of NR4A1.
10.11817/j.issn.1672-7347.2015.04.001
- Author:
Ningning YAN
1
;
Jun LI
;
Xiaojuan CHEN
;
Yongheng CHEN
;
Lin CHEN
;
Zhuchu CHEN
Author Information
1. Laboratory of Structural Biology, Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.
- Publication Type:Journal Article
- MeSH:
Electrophoresis, Polyacrylamide Gel;
Nuclear Receptor Subfamily 4, Group A, Member 1;
chemistry;
Recombinant Fusion Proteins;
chemistry
- From:
Journal of Central South University(Medical Sciences)
2015;40(4):345-350
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To express and purify NR4A1-DNA binding domain (DBD) protein of nuclear receptors.
METHODS:The fusion protein PET28a-NR4A1-DBD was constructed and purified with the nickel affinity chromatography, cation-exchange chromatography and gel filtration chromatography.
RESULTS:The protein PET28a-NR4A1-DBD was mostly soluable at 24 °C. A total of 2-3 mg/L pure NR4A1 proteins were yielded in bacterial culture and the purity for final fractions of NR4A1-DBD protein were great than 95% by SDS-PAGE analysis.
CONCLUSION:Nickel affinity chromatography is effective to purify protein. The protein purity can be further improved by the following methods including cation-exchange chromatography and gel filtration chromatography.
