[Expression and purification of DNA binding domain of NR4A1].
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.


CONCLUSIONS
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.
Laboratory of Structural Biology, Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.
Ningning
N
Yan
Jun
J
Li
Xiaojuan
X
Chen
Yongheng
Y
Chen
Lin
L
Chen
Zhuchu
Z
Chen
D004591:Electrophoresis, Polyacrylamide Gel; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1 / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry
D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; D011993:Recombinant Fusion Proteins