Expression and characterization of recombinant Müllerian Inhibiting Substance (MIS) using the Bombyx Mori Baculovirus – insect cell expression system

Abstract The Müllerian inhibiting substance (MIS) is a glycoprotein that belongs to the transforming growth factor-β family and is known for its crucial roles in embryonic development and cellular differentiation. Attempts to produce recombinant MIS using bacterial and yeast expression systems have faced challenges, including insufficient post-translational modifications and low expression efficiency. In this study, the MIS gene was successfully cloned into the pBacPAK8 transfer vector and expressed using the Bombyx mori baculovirus - insect cell expression system. Co-transfection of BMN1 insect cells with the recombinant plasmid and baculoviral DNA led to the successful generation of recombinant baculoviruses. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis and Western blot analyses of lysates from infected cells confirmed the expression of a ~70 kDa MIS protein with significant antigenicity. To the best of our knowledge, this is the first report describing the recombinant production of human MIS protein using the B. mori baculovirus system. This expression platform provides a promising approach for synthesizing structurally complex human proteins and lays the groundwork for future research and biomedical applications.

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