Figure 1
Operons and genomic regions deleted in P . putida KT2440 to construct a cell factory strain. (A) Position of the eleven gene(s)/regions deleted in wild-type P. putida KT2440 indicated in the physical map of the chromosome. (B) Roadmap for the construction of strains EM42 and EM383. Relevant genes are depicted in the order in which they were eliminated (see also Additional file 1: Table S1). (C) Electrophoresis of the diagnostic PCR amplifications to confirm the deletions. The flanking lanes (M) correspond to a DNA ladder [500-bp Molecular Ruler EZ Loadâ„¢ (Bio-Rad Corp., Berkeley, CA, USA)], and lanes identified as Ï• are negative controls, i.e., samples without DNA template. The photograph shows the products resulting from PCR amplifications of [i] an internal gene within prophage 1, KT2440 (lane 1) and EM383 (lane 2); [ii] an internal gene of prophage 2, KT2440 (lane 3) and EM383 (lane 4); [iii] an internal gene of prophage 3, KT2440 (lane 5) and EM383 (lane 6); [iv] an internal gene of prophage 4, KT2440 (lane 7) and EM383 (lane 8); [v] an internal gene of the hsdRMS operon, KT2440 (lane 9) and EM383 (lane 10); [vi] the TS1-TS2 region of recA, KT2440 (lane 11) and EM383 (lane 12); [vii] an internal gene of the Tn7-like operon, KT2440 (lane 13) and EM383 (lane 14); [viii] the TS1-TS2 region of endA-1, KT2440 (lane 15) and EM383 (lane 16); [ix] the TS1-TS2 region of endA-2, KT2440 (lane 17) and EM383 (lane 18); [x] an internal gene of the flagellar operon, KT2440 (lane 19) and EM383 (lane 20); and [xi] an internal gene of the Tn4652 operon, KT2440 (lane 21) and EM383 (lane 22). The details of primers sequence used in these amplifications are given in Additional file 1: Table S2.