| Summary: | 碩士 === 國立中興大學 === 食品暨應用生物科技學系所 === 107 === Myostatin (MSTN), also known as growth and differentiation factor 8 (GDF-8), is a member of the transforming growth factor β (TGF-β) superfamily. It plays a key role in not only negative regulation of development and differentiation of skeletal muscle but also adipogenesis. Blocking myostatin activity might be useful for feed, dietary supplement or muscle disease treatment.
In this study, we constructed some expression systems of recombinant Myo epitope fusion polypeptide, including L. lactis NZ9000 system, B. subtilis WB800 system and E. coli BL21(DE3) system. We successfully expressed recombinant Myo epitope fusion polypeptide in E. coli BL21(DE3) system and GRAS B. subtilis WB800 system after many tries of various plasmids construction.
In animal mode, we vaccinated mice by orally fed with purified Myo epitope fusion polypeptide from E. coli BL21(DE3) or cell pellet from B. subtilis as immuno-blocking antigen (IMBA). In addition, we also examined recombinant ganoderma lucidum immunomodulatory protein Ling Zhi-8 (LZ-8) with purified Myo epitope fusion polypeptide as IMBA. Results showed that the compound Myo epitope fusion polypeptide with rLZ8 exhibited best effect to stimulate total IgA, myostatin-specific IgG1, IgG2a and promotion of TH1 immune response. Strikingly, the combined compound could lower the body weight of obese mice to normal mice level. Based on this fact, we confirmed recombinant Myostatin immuno-blocking antigen bioactivity and immune-modulation of rLZ-8. Based on our results, future studies can be performed to improve myostatin immuno-blocking antigen design, and develop more efficiency GRAS/Food Grade myostatin immuno-blocking antigen to apply in feed, healthy food or medical industry.
|
|---|
