ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2010, Vol. 41 ›› Issue (3): 279-285.doi:

• 遗传繁育 • Previous Articles     Next Articles

Transient Transfection Factors for Expression of Recombinant Somatostatin Plasmid pEGS/2SS in Cultured HeLa Cells

HE Xiaohong 1,4, CAO Shaoxian 2, MAO Dagan 1, YANG Liguo 3*
  

  1. 1.College of Animal Science and Technology, Nanjing Agricultural University,NanJing 210095, China; 2.Institute of Animal Science, Jiangsu Academy of Agricultural Sciences,Nanjing 210014, China; 3.College of Animal Science and Technology,Huazhong Agricultural University, Wuhan 430070, China； 4.Institute of Animal Science,Chinese Academy of Agricultural Sciences, Beijing 100193,China
  • Received:2008-11-25 Revised:1900-01-01 Online:2010-03-24 Published:2010-03-24
  • Contact: YANG Liguo

PDF (PC)

649

Abstract: We aimed to investigate the factors infecting transfection efficiency of somatostatin recombinant plasmid pEGS/2SS introducting into HeLa cells, make preparations for advanced study on somatostatin gene vaccine mechanism and effect of action. Four parameters including transfection reagent (lipofectamineTM) concentration, DNA concentration, cells cultured time after transfection and DNA extracting techniques were analyzed and optimized, and transfection efficiency was evaluated by the percentage of transfected cells with green fluorescence under fluorescence microscope. Based on the optimized lipofectamineTM and DNA concentration, 34.02% transfected HeLa cells could express green fluorescence 72 h after transfection when the concentration of lipofectamineTM and DNA was individually 6 μg and 1 μg; Based on the optimized cells cultured time after transfection and DNA extracting techniques, 17.88% transfected HeLa cells could express green fluorescence protein with Plasmid Maxi Kit extracting DNA 48 h after trasfection. Optimal transfection conditions were determined with four parameters, 17.88% HeLa cells could express green fluorescence protein with Plasmid Maxi Kit extracting DNA, 1 μg DNA and 6 μg lipofectamineTM 48 h after transfection.