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Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-547646

RESUMEN

Objective In order to research the best fermentation conditions of recombinant bacteria pThioHisA-B1/DH5? which contains gene of esterase B1.Methods Recombinant bacteria pThioHisA-B1/DH5? was induced in different culture media, induction temperatures, different inductor concentrations, different induction time in order to obtain the best expression effect.The expression products of different conditions were detected by SDS-PAGE and thin-layer gel scanning analysis.Results Based on the experiments, the optimized fermentation condition were that the culture medium was TB, inducing temperature was 28 ℃ and inducing time was 6 h,the concentration of IPTG was 0.4 mmol/L.The expression quantity of target protein attained 40.3% in total bacterial protein, and the soluble fusion protein accounted for 66.2% in total target proteins at the optimum conditions.Conclusion The optimized fermentation conditions of expressing fusion protein in recombinant bacteria have been found and in the conditions, soluble esterase B1 fusion protein of high yield can be obtained.

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