Reconstruction and expression of recombinant adenovirus co-expressing BMP2 and BMP9

Objective To reconstruct and identify the recombinant adenovirus co-expressing BMP2 and BMP9.Method The genes of BMP2 and BMP9 were amplified from AdEasy vector by PCR and sub-cloned into pASG2 vector.The co-expression shuttle plasmid pASG2-BMP2,9 was confirmed by restriction endonuclease digestion,PCR and gene sequencing,then pASG2-BMP2,9 was electro-transducted into competent AdEasier cells to acquire recombinant adenovirus plasmid.Then,the recombinant vector was transfected into HEK293 cells and high-titer recombinant adenovirus(AdBMP2,9) was gained after rounds of amplification.The expression and bone induction capacity of AdBMP2,9 was observed in C3H10 cells.Result AdBMP2,9 was constructed successfully and the virus titer was 1010 IU/mL after amplification.AdBMP2,9 could express and induce alkaline phosphatase activity in C3H10 cells.Conclusion The recombinant adenovirus co-expressing BMP2 and BMP9 was constructed successfully,which provides a useful tool for bone tissue engineering.