TY - JOUR JF - Res-Mol-Med JO - Res Mol Med (RMM) VL - 9 IS - 1 PY - 2021 Y1 - 2021/2/01 TI - Cloning, Expression, and Purification of Recombinant Mouse Interferon-γ TT - N2 - Background: Interferon-gamma [IFN-γ) is the most important cytokine in the immune system. This protein has been expressed in bacterial cells. However, bacterial cloning is not an easy task. We aimed to clone, express, and purify recombinant mouse IFN-γ and overcome problems in favor of commercial purposes. Materials and Methods: To amplify the gene product for cloning, we primarily designed two specific primers for the target gene. Following PCR amplification, the amplicon was inserted into the pET-21b[+) vector. The E. coli BL21 [DE3) CodonPlus strain was chosen for the expression of the target gene. Finally, the expressed recombinant mouse IFN-γ was assessed through the western blotting method. Results: We performed a cloning process and produced recombinant mouse IFN-γ in an optimal condition. We also noticed that monomeric protein could be transformed to a homodimeric structure which can be observed using the SDS PAGE [SDS-polyacrylamide gel electrophoresis) and western blotting. Conclusion: Experimental conditions strongly affect the large-scale cloning procedures required to be optimized in each laboratory. The expressed recombinant mouse IFN-γ described here is appropriate for commercial purposes. SP - 1 EP - 10 AU - Badiee Kheirabadi, Seyedeh Elham AU - Mashayekhi, Kazem AU - Moghadam, Malihe AU - Mousavi, Mohammad Javad AU - Sankian, Mojtaba AD - Department of Hematology, Faculty of Allied Medicine, Bushehr University of Medical Sciences, Bushehr, Iran. KW - Cloning KW - E. coli BL21 (DE3) Codon-Plus KW - pET21-b (+) vector KW - Interferon gamma KW - Recombinant protein UR - http://rmm.mazums.ac.ir/article-1-398-en.html DO - 10.32598/rmm.9.1.1 ER -