[Construction of adeno-associated virus vector containing ANG-1 gene and its expression in pig mesenchymal stem cells].
OBJECTIVE
To construct recombinant adeno-associated virus (rAAV) vector containing angiopoietin-1 (ANG-1) gene and to express the ANG-1 in targeting cells.


METHODS
ANG-1 cDNA was obtained from human spleen by RT-PCR and was inserted into AAV vectors to form rAAV ANG-1, the virus stocks in high titer were harvested. The rAAVANG-1 and rAAV GFP were transferred into pig mesenchymal stem cells and the expression of ANG-1 was detected by Western blot.


RESULTS
The cloned ANG-1 cDNA was 1515bp in length which was in accordance with that reported previously. Titration of rAAVANG-1 stock was 9 X 10(11)v.g/ml. The expression of ANG-1 gene was detected in transfected cells. Forty-eight hours after rAAV GFP was transfected into mesenchymal stem cells, 55% cells expressed GFP.


CONCLUSIONS
The constructed rAAV ANG-1 vector has successfully transfered and expressed in pig mesenchymal stem cells.
ZHU
Cheng-chu
CC
Department of Cardiothoracic Surgery, Taizhou Hospital, Linhai 317000, China.
CHEN
Shi-lin
SL
LIU
Yu-qing
YQ
TANG
Li-jiang
LJ
BAO
Wei-guang
WG
D042683:Angiopoietin-1 / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D013552:Swine; D014162:Transfection
D042683:Angiopoietin-1; D018076:DNA, Complementary; D011994:Recombinant Proteins