Construction of BMP7 Recombinant Adenovirus and Expression of BMP7 in Transfected Rabbit Nucleus Pulposus Cells

Objective To construct a recombinant adenovirus carrying human BMP7 gene which is transfected into rabbit nucleus pulposus cells and observe its expression.Methods ⑴ BMP7 adenovirus was recombined with a series of restrictive enzyme treatment of the pcDNA3.1(+)-BMP7,large-scale virus was prepared by using HEK 293 cells and viral titer was determined.⑵ Twenty New Zealand white rabbits were chosen and 15 rabbits were in experimental group while 5 rabbits in the control group.Ad-BMP7,Ad-GFP carrying adenovirus 6×10~6 pfu was injected directly into rabbit nucleus pulposus of lumbar discs in experimental group and the control group respectively.Self control group remained for reference.The expression of BMP7 was detected by RT-PCR and immunohistochemistry(at 1st week,2nd week and 4th week).Results ⑴ BMP7 was able to be cut by the restrictive enzyme of PI-SceⅠand I-CeuⅠfrom adenovirus BMP7 recombinant and BMP7 mRNA expression was observed by PCR,so it came true that we had constructed real adenovirus BMP7.⑵ BMP7 gene was expressed effectively in the experimental group.There was no significant diffrence among the groups at the 1st week,the 2nd week and the 4th week(P0.05).Conclusion Construction of BMP7 adenovirus and its expression lays the basis for the further research in the bioactivity of the nucleus pulposus cells.