[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.
Liu
S
S
Molecular Medicine Center, Cardiovascular Institute, CAMS and PUMC, Beijing 100037.
Liu
D
D
Ding
J
J
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004622:Embryo, Mammalian; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D005263:Femoral Artery / Q000378:metabolism*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D007668:Kidney / Q000166:cytology; D008297:Male; D010957:Plasmids / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics*; D014947:Wounds and Injuries / Q000378:metabolism
D018076:DNA, Complementary; D004792:Enzyme Precursors; D011994:Recombinant Proteins; D014568:Urokinase-Type Plasminogen Activator; C067551:saruplase