Expression, Purification and Activity Analysis of Arabidopsis-dioxygenase 1

doi:10.7668/hbnxb.2008.03.003

Abstract

Abstract: Recombinant DOXI was cloned into pMAL-c4x expression vector and expressed in E. coli T7 ExpressCompetent E. cola and BL21( DE3) RLPL codon⁺.Purified using amylose resin column, a fusion protein about 114 kD wasdetected by SDS-PAGE in the IPTG induced recombinant BL21(DE3) RIPL codon⁺ strain，and it's yield acounts for3. 7% of the total bacterial proteins, the purity of recombinant proteinwas about 45%.The soluble fusion protein had un-detectable peroxidase activity by the guaiacol methol;The alpha dioxygenase activity of the purified protein was tested us-ing 2, 4 DNP，result showed that the soluble fusion protein had detectable alpha-dioxygenase activity.

Key words: Alpha-dioxygenase 1, Recombinant expression, 2,4-DNP, Affinity purification, Activity

CLC Number:

Q786

ZHANG Xiao-mei, HAN Nian-fa, ZHANG Mei-xiang, LI Guang-lu, FAN San-hong, GUO Ai-guang,. Expression, Purification and Activity Analysis of Arabidopsis-dioxygenase 1[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(3): 9-12. doi: 10.7668/hbnxb.2008.03.003.

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