Synechocystis Fe superoxide dismutase gene confers oxidative stress tolerance to Escherichia coli |
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Authors: | Bhattacharya Jyotirmoy GhoshDastidar Krishnarup Chatterjee Anirban Majee Manoj Majumder Arun Lahiri |
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Affiliation: | Plant Molecular and Cellular Genetics, Bose Institute (Centenary Building), P-1/12, CIT Scheme VII M, Kolkata 700 054, India. |
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Abstract: | The superoxide dismutase (SOD) gene (slr 1516) from the cyanobacterium Synechocystis sp. PCC 6803 was cloned and overexpressed in Escherichia coli BL 21 (DE3) using the pET-20b(+) expression vector. E. coli cells transformed with pET-SOD overexpressed the protein in cytosol, upon induction by isopropyl beta-D-thiogalactopyranoside (IPTG). The recombinant protein was purified to near homogeneity by gel filtration and ion-exchange chromatography. The SOD activity of the recombinant protein was sensitive to hydrogen peroxide and sodium azide, confirming it to be FeSOD. The pET-FeSOD transformed E. coli showed significantly higher SOD activity and tolerance to paraquat-mediated growth inhibition compared to the empty vector transformed cells. Based on these results it is suggested that overexpression of FeSOD gene from a heterologous source like Synechocystis sp. PCC 6803 may provide protection to E. coli against superoxide radical-mediated oxidative stress mediated by paraquat. |
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Keywords: | Synechocystis sp. PCC 6803 Iron superoxide dismutase FeSOD Paraquat Oxidative stress Escherichia coli |
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