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Figure 4 | BMC Biotechnology

Figure 4

From: Regulated expression of a transgene introduced on an oriP/EBNA-1 PAC shuttle vector into human cells

Figure 4

Transfection efficiencies measured in 293T and SK-BR-3 cells. A) DNA from each of the constructs pPAC7, pPAC7-39 and pPAC7-CDH3 was digested with NotI and separated on a BioRad CHEF-mapper. During construction of pPAC7-39 one of the NotI restriction sites got deleted and thus giving a linearised fragment when digested with NotI. Selected fragments in the Low Range PFG Marker are indicated. B) Diagram showing the fraction of viable, EGFP-expressing 293T and SK-BR-3 cells measured by flow cytometry the day after the transfection with the following constructs: pPAC7, pPAC7-39 and pPAC7-CDH3. One μg of DNA from each construct was used for transfection, and the figure shows the transfection frequencies normalized with respect to the molar concentrations of plasmid DNA used in each transfection. For the 293T cell line, each bar represents three independent experiments all performed in triplicate. For the SK-BR-3 cell line each bar represents two separate experiments, both performed in triplicates. Standard deviations are indicated. C) Representative flow diagrams showing a 10-fold decrease in EGFP intensities in the 293T cell line with increasing plasmid size. Constructs used are: pPAC7, pPAC7-39 and pPAC7-CDH3.

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