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. 2014 Jun 30;15(9):918–926. doi: 10.1111/mpp.12154

Figure 1.

figure

Confirmation of four independent ToxA knockout strains. (a) Reverse transcription‐polymerase chain reaction (RT‐PCR) detection of ToxA transcript (PCR product size of 393 bp) in the four knockout strains and wild‐type (WT), as visualized by agarose gel electrophoresis. Act1 was included as a positive control (PCR product size of 150 bp). (b) Correct integration of the ToxA gene deletion cassette at 5′ and 3′ ends in four replicates of the knockout strains. PCR product sizes for the 5′ and 3′ amplicons are 1.7 kb and 1.6 kb, respectively. (c) Phleomycin resistance cassette (PhleoR) copy number as detected by quantitative PCR (qPCR). Error bars depict standard deviation. (d) Sodium dodecylsulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) of culture filtrate proteins. Open arrows indicate ToxA (13.2 kDa). Two independent wild‐type (WT) culture filtrate samples were analysed.