Expression and Characterization of Recombinant Human Anti-thrombin in Saccharomyces Cerevisiae
نویسندگان
چکیده
Objective: The purpose of this study was to express and purify recombinant anti-thrombin in Saccharomyces cerevisiae BY4741 expression system. Methods: cDNA of human anti-thrombin gene was amplified with specific primers and cloned into pYES2/CT vector with His-tag for easier purification using Ni2+ Results: Protein was purified to homogeneity and was confirmed as anti-thrombin, using SDS-PAGE and western blotting. We characterized recombinant anti-thrombin and compared it with human plasma-derived anti-thrombin. -NTA chromatography. Conclusion: Anti-thrombin III isolated from the culture media of Saccharomyces cerevisiae BY4741 was biologically active, as could be shown by progressive anti-thrombin activity and heparin cofactor activity.
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