[Expression and purification of DNA binding domain of NR4A1]

Ningning Yan; Jun Li; Xiaojuan Chen; Yongheng Chen; Lin Chen; Zhuchu Chen

doi:10.11817/j.issn.1672-7347.2015.04.001

[Expression and purification of DNA binding domain of NR4A1]

Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2015 Apr;40(4):345-50. doi: 10.11817/j.issn.1672-7347.2015.04.001.

[Article in Chinese]

Authors

Ningning Yan¹, Jun Li, Xiaojuan Chen, Yongheng Chen, Lin Chen, Zhuchu Chen

Affiliation

¹ Laboratory of Structural Biology, Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.

PMID: 25931227
DOI: 10.11817/j.issn.1672-7347.2015.04.001

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Electrophoresis, Polyacrylamide Gel*
Nuclear Receptor Subfamily 4, Group A, Member 1 / chemistry*
Recombinant Fusion Proteins / chemistry

Substances

Nuclear Receptor Subfamily 4, Group A, Member 1
Recombinant Fusion Proteins