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Fig. 2 | Biotechnology for Biofuels

Fig. 2

From: Mutant generation by allelic exchange and genome resequencing of the biobutanol organism Clostridium acetobutylicum ATCC 824

Fig. 2

Isolation of in-frame deletion mutants of spo0A and cac824I. Schematic representations of the wild-type and mutant genomes in the targeted regions are shown above each electrophoretogram, together with the relative position of the two primers used and the extent of the Left Homology Arm (LHA) and Right Homology Arm (RHA) employed to mediate recombination. In both: MW, 2-log DNA marker (NEB) molecular weight marker; ‘W’, is a PCR with no DNA; ‘P’, is a PCR using the plasmid only, and; ‘WT’ is the wild-type strain CRG1268. a PCR screening of four FCR colonies (labelled lanes 14) using primers Cac-spo0A-sF2 (1) and Cac-spo0A-sR2 (2). Expected fragment sizes are 1679 bp in the deletion mutant and a 2114 bp fragment in the wild-type, respectively. Accordingly, labelled lane WT is wild-type, lanes 1, 3 (very weak band) and 4 are mutants. b PCR screening of seven FCR colonies (labelled lanes 1–7) using primers Cac-1501-sF2 (1) and Cac-1504-sR1 (2). Expected fragment sizes are 1611 bp in the deletion mutant and a 2319 bp fragment in the wild-type, respectively. Accordingly, labelled lanes 4–7 are wild-type, lane 2 is a mutant, and lanes 1 and 3 are a mixture of mutant and wild-type

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