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Construction of adeno-associated virus vector containing ANG-1 gene and its expression in pig mesenchymal stem cells

Bao Yu-guang

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Abstract

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 9×1011v.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.Conclusion: The constructed rAAV ANG-1 vector has successfully transfered and expressed in pig mesenchymal stem cells.

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What this paper is about

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 9×1011v.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.Conclusion: The constructed rAAV ANG-1 vector has successfully transfered and expressed in pig mesenchymal stem cells.

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Available abstract

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 9×1011v.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.Conclusion: The constructed rAAV ANG-1 vector has successfully transfered and expressed in pig mesenchymal stem cells.

Key concepts: Adeno-associated virus, Molecular biology, Mesenchymal stem cell, Complementary DNA, Transfection, Biology, Virology, Viral vector

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