Improved Soluble Expression in Escherichia coli and Easily Purified Recombinant Human Bone Morphogenetic 7-2 Fusion Protein

Background: Bone morphogenetic protein (BMP) is a cysteine-rich growth factor and plays a key role in early bone tissue development and bone defect repair. However, the low yield, high cost and complicated process in BMP significantly limit its clinical application. Objective: In this study, we developed an efficient method for soluble expression and preparation of recombinant human bone morphogenetic 7-2 fusion protein (rhBMP7-2) and determined its molecular weight and biological activity. Methods: The fusion gene for rhBMP-2 and rhBMP-7 was inserted into the pET-ELP expression vector. Correct DNA sequence was confirmed, the rhBMP7-2-ELP was transformed into Escherichia coli strain BL21 (DE3), and the rhBMP7-2 was produced in the recombinant E. coli. Recombinant BMP7-2 purify was identified using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). The cell proliferation and biological activity of rhBMP7-2 were measured by Cell Counting Kit-8 and Alkaline Phosphatase assay using C2C12 cells, respectively. Results: The result of digestion of NdeI, BamHI and XhoI enzymes showed that the rhBMP7-2-ELP was correctly constructed. The recombinant BMP7-2 was successfully expressed in soluble form; the purified rhBMP7-2 showed biological activity and significantly promoted cell proliferation and differentiation in a dose-dependent manner. Conclusion: The rhBMP7-2 fusion protein with osteogenic activity was prepared through a low-cost and time-efficient method. Our preparation method presents the potential to be applied to the large-scale production of rhBMP7-2 and is expected to play a significant role in clinical treatment.