Figure 5.

Quantitation of recombined locus frequency. A, shows PCR analyses of the parental (WT) and Δppdk/Δpepck/^RNAiGPDH cell lines using primers designed for DNA amplification of the chimeric FRDg-m2 gene (upper panel) or the FBPase gene for normalisation (lower panel). The fraction of recombined loci was determined by calculating the relative difference between the two linear regressions of FRDg-m2 PCR signal (gel scan) as a function of the amount of input genomic DNA (B) (mean of three independent experiments). The value obtained for the FRDg-m2 PCR signal with 10 μg ml⁻¹ of Δppdk/Δpepck/^RNAiGPDH genomic DNA, highlighted by an asterisk, was excluded from the trend line.