Cloning, Expression, and Purification of Recombinant Mouse Interferon-γ

Background: Interferon-gamma [IFN-γ) is the most important cytokine in immune system. This protein has been expressed bacterial cells. However, cloning not an easy task. We aimed to clone, express, and purify recombinant mouse IFN-γ overcome problems favor of commercial purposes. Materials Methods: To amplify gene product for cloning, we primarily designed two specific primers target gene. Following PCR amplification, amplicon was inserted into pET-21b[+) vector. The E. coli BL21 [DE3) CodonPlus strain chosen expression Finally, assessed through western blotting method. Results: performed a process produced optimal condition. also noticed that monomeric could be transformed homodimeric structure which can observed using SDS PAGE [SDS-polyacrylamide gel electrophoresis) blotting. Conclusion: Experimental conditions strongly affect large-scale procedures required optimized each laboratory. described here appropriate