The expression and purification of human histone demethylase JARID1B and in vitro demethylation activity analysis

Objective To obtain high-quality truncated human histone demethylase JARID1B(jumonji AT-rich interactive domain 1B) in large amount.Methods We used Bac-to-Bac baculovirus expression system to express proteins followed by Ni-NTA affinity chromatography,ion exchange and gel filtration.Matrix-assisted laser desorption ionization time of flight mass spectrometer(MALDI-TOF) was employed to analyse the demethylation activity.Results JARID1B1-825,a construct lack of the C-terminal domain,was able to be expressed in Hi5 insect cells in great quantity.The yield was 10 mg protein per liter insect cell culture.In vitro demethylase activity analysis of JARID1B1-825 showed that this truncated JARID1B1-825 possessed the demethylation activity.Conclusions Large amount of JARID1B with hight-quality was obtained.This research will be useful for future structural and functional studies of JARID1B and related drug screening.