Construction of adeno-associated virus vector containing ANG-1 gene and its expression in pig mesenchymal stem cells.
- Author:
Cheng-chu ZHU
1
;
Shi-lin CHEN
;
Yu-qing LIU
;
Li-jiang TANG
;
Wei-guang BAO
Author Information
- Publication Type:Journal Article
- MeSH: Angiopoietin-1; biosynthesis; genetics; Animals; DNA, Complementary; genetics; Dependovirus; genetics; metabolism; Genetic Vectors; genetics; Humans; Mesenchymal Stromal Cells; metabolism; Recombinant Proteins; biosynthesis; genetics; Swine; Transfection
- From: Journal of Zhejiang University. Medical sciences 2009;38(4):370-376
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct recombinant adeno-associated virus (rAAV) vector containing angiopoietin-1 (ANG-1) gene and to express the ANG-1 in targeting cells.
METHODSANG-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.
RESULTSThe 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.
CONCLUSIONThe constructed rAAV ANG-1 vector has successfully transfered and expressed in pig mesenchymal stem cells.
