Expression and purification of DNA binding domain of NR4A1 / 中南大学学报(医学版)

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.