Figure 2

Identification of acsA as a down-expressed gene in mutant devoid of the LuxR-type regulator. A - Construction of V. vulnificus mutant defective in the LuxR-type transcription factor by using two sets of primers (indicated by horizontal arrows with the primer names listed in Additional file 4: Table S2) to delete VVMO6_00196. A bar represents the length of DNA equivalent to 500 bp; B - Deletion of the corresponding gene was examined by PCR using a pair of primers, luxRupF and luxRdownR; C - Confirmation of the deletion mutant. No production of the LuxR-type transcription factor in the mutant was confirmed by western blot analysis using polyclonal antibodies raised against recombinant protein of the LuxR-type transcription factor; D - Real-time PCR assay measuring the relative transcript level of acsA in mutant defective in the LuxR-type transcription factor. Relative transcript levels of acsA in wild-type and mutant strains were estimated as described in Figure 1. Data are presented as the mean ± standard deviation from three independent experiments. Statistical analyses were performed using Student t-test to evaluate the statistical significance of the results. A datum with P <0.01 is indicated with two asterisks. SM indicates DNA or protein size markers.