Optimization of transfection conditions for tranfecting pAcGFP-bFADD recombinant plasmid to Hela cell mediated by lipofectamine 2000.

pAcGFP-bFADD plasmid with AcGFP report gene was transfected to Hela cell mediated by lipofectamine 2000, and the transfection condition was optimized by adjusting transfection time and concentrations of plasmid DNA and liposome.Twelve hours after transfected, positive cells count was made under fluorescence microscope, and the cell viability was detected by MTT.Results indicated that perfect cell transfection efficiency(62.3% ) was obtained on the condition of the complex of 2.4 μg pAcGFP-bFADD and 3.0 μL lipofectamine 2000 transfecting Hela cell after 6 h, meanwhile, its cell viability could reach 83%.When the concentration of plasmid DNA and liposome increased to 3.2 μg and 6.0 μL respectively, the complex would bring higher toxicity, the inhibition rate reached 42.98%, the difference was extremely significant(P 0.01) compared with control group.After condition optimization, AcGFP-bFADD fusion gene got the expression peak 72 h after transfection.The green fluorescent positive cells ratio could reach 64%.