Production and purification of firefly luciferase inEscherichia coli
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A genetic recombinant stain ofE.coli were induced to express and secrete firefly luciferase. Cells, when broken by freeze/thawing, gave about 2% of the total soluble protein as luciferase. The luciferase was purified with ammonium sulphate fractionation and gel filtration chromatography giving a luciferase product with high specific activity (106 light units/mg protein). SDS-PAGE of this product showed two active bands at 54 and 50 kDa, which corresponded to the luciferases with and without a signal peptide on their N-terminals. The yield was more than one mg purified enzyme per 100 ml of fermentative liquid.
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