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›› 2007, Vol. 27 ›› Issue (3): 289-292.doi: 10.7525/j.issn.1673-5102.2007.03.008

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Cloning and Expression of Gene which Encode SOD of Nostoc flagelliforme in E. coli

WANG Ying;CHEN Li-Ping;CHEN Xiao;ZHANG Xiu;YU Jing;WANG Quan-Xi*   

  1. College of Life and Environment Sciences,Shanghai Normal University,Shanghai 200234
  • Received:1900-01-01 Revised:1900-01-01 Online:2007-05-20 Published:2007-05-20
  • Contact: WANG Quan-Xi

Abstract: SOD gene was cloned from Nostoc flagelliforme and the amino acid sequence is 97% identical to that of Nostoc commune published. The gene was inserted into a constructed E.coli expression plasmid pET-sod and the plasmid was transferred into expressing host BL21. Induced by 1 mmol·L-1 IPTG,the recombinant SOD of Nostoc flagelliforme was accumulated to a very high percent and the protein is exist as soluble protein. SDS-PAGE analysis revealed that the molecular weight of expression SOD of Nostoc flagelliforme was approximate 22 kd. Purified by Ni2+-resin column,the specific enzymatic activity was measured using NBT method and the calculated result of this purified enzyme is 2 550 U·mg-1. It was also found that after high temperature stress at 60℃ for 90 min,the specific enzymatic activity still retains 85%.

Key words: Nostoc flagelliforme, gene cloning of SOD, induced expression, activity measurement of SOD