Objective       To construct an adenovirus vector of rat deltex-1 gene and then investigate its expression in rat bone marrow mesenchymal stem cells (bMSCs).      Methods      After deltex-1 gene was amplified by high-fidelity PCR

the product was subcloned to the pAdTrack-CMV adenovirus vector after sequencing

and recombined in the BJ5183 bacterium. The correct recombinant were picked and then packaged to virus in FT293 cells. Then the virus was purified with CsCl

2

and its titer was determined. At last

the

expression of deltex-1 gene was determined by detecting the contents of total RNA and protein of the bMSCs with the target gene by PCR and Western blotting respectively.      Results      Rat deltex-1 gene fragment of 1 883 bp was successfully amplify

and constructed in the reconstituted adenovirus shuttle vector of 4.5 kb

and the titer of the vector was 1.23×10

11

IU/ml. Flow cytometric analysis showed cell-surface antigens for rat bMSCs were CD90 99.57%

CD44 85.66% and CD45 0.35%. The efficiency of transfection was 60% to 70% under fluorescent microscope. RT-PCR and Western blotting suggested that the expression of deltex-1 of transfected MSCs was obviously higher than controls. All of these reveal that the transfection of deltex-1 into rat bMSCs was successful.      Conclusion      The adenovirus vector of deltex-1 is successfully constructed and the bMSCs infected by the virus express the target gene at a high level.