[Construction and in vivo expression of the recombinant adenovirus containing human pro-UK cDNA].

OBJECTIVE

To construct the recombinant adenovirus which contained human pro-UK cDNA and to provide experimental basis for clinical gene therapy.

METHODS

The recombinant plasmid pCA14-pro-UK and the plasmid pJM17 were cotransfected into cultured 293 cells and the replication-deficient adenovirus were harvested, identified, propagated and titered. The recombinant virus were then transferred into balloon-injured femoral arteries of rabbits and the pro-UK expression was detected by immunohistochemistry and Western Blot.

RESULTS

PCR amplification and in vitro transfection of A549 cells confirmed that pro-UK had been inserted into adenovirus genome correctly, and immunohistochemistry and Western blot analysis showed that pro-UK expressed evidently in the balloon-injured sites of the rabbit femoral arteries.

CONCLUSIONS

The recombinant adenovirus that constructed could express pro-UK cDNA in vivo successfully.