Abstract:To produce recombinant human MBD4 protein, the ORF of MBD4 cDNA was inserted into pGEX-6P1 expression vector fused in frame with an upstream GST gene. The expression plasmid was transformed into E.coli BL21(DE3) strain and protein expression was induced by IPTG. The GST-MBD4 fusion protein was purified from cell lysates using glutathione Sepharose 4B affinity chromatography. The GST tag was removed from fusion protein by site-specific Prescision protease cleavage. MBD4 protein was purified through Mono Q anion exchange to more than 94% homogeneity. The purified protein has both methylated DNA binding and DNA glycosylase activity.
李伟. 重组人MBD4蛋白在大肠杆菌中的表达、纯化及活性分析[J]. 中国生物化学与分子生物学报, 2006, 22(12): 979-983.
LIWei. Purification and Characterization of Recombinant Human MBD4 Protein Expressed in E.coli . Chinese Journal of Biochemistry and Molecular Biol, 2006, 22(12): 979-983.