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Phosphorylation-independent activation of the atypical response regulator NblR, by D. Ruiz, P. Salinas, M. L. Lopez-Redondo, M. L. Cayuela, A. Marina and A. Contreras

Microbiology vol. 154, part 10, pp. 3002 - 3015

Fig S1. Strategy for generation and analysis of strains WT-RCS3 and NblRD57A-RCS3. (A) The integration of the CS3 cassette (hatched bars) in the intergenic region (solid bar) between nblR and the downstream ORFs Synpcc7942_2306 (open arrows) by homologous recombination is shown schematically. Flanking chromosome regions are represented by dotted lines and plasmid sequences by a continuous line. Depending on specific crossover sites two alternative nblR alleles can be generated. Mutant (nblR_D57A) and wild-type strains differ at the indicated PvuI site. (B) Schematic representation of the allele structure in strains WTRCS3 and NblRD57A-RCS3. Relevant PvuI sites are shown. Positions of primers used to verify allele structure are indicated by black arrows. (C) PCR analysis of WT-RCS3 (lane 1), NblR_D57A-RCS3 (lane 2) and Synechococcus sp. PCC7942 (lane 3) using primers NblR-1F (1F) and CS3-2R (2R). (D) PvuI digestion of the PCR fragment generated with primers 1F and NblR-1R (1R). Lane numbers as in (C). M: size marker λ HindIII+EcoRI. L: DNA 100 bp ladder (Fermentas). [PDF] (312 kb)

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