autoinducer-deficient Vibrio cholerae strain transformed with library of random Vibrio cholerae genomic fragments fused to a promoterless luciferase cassette. Verification of Quorum sensing (QS) regulation: wild-type Vibrio cholerae El and QS mutants are transformed with each candidate reporter plasmid, each candidate exhibits similar expression pattern in a luxO D47E strain (response regulator luxO) and a hapR strain (QS transcription factor HapR) that both simulate low cell density, indicating expression of each target gene is dependent on LuxO-P as well as HapR. VCA0681 is overexpressed in Vibrio cholerae luxO D47Eel: reduction of reporter expression. Deletion of either VCA0681 from the Vibrio cholerae El genome does not alter biofilm formation (transcriptional activator vps expression). Absolute level of vps expression in the (HapR minus) Vibrio cholerae El strain 100 times higher than that of wild-type (HapR minus) Vibrio cholerae C1 strain
commercial synthetic gene PA4108, DNA and amino acid sequence determination and analysis, recombinant overexpression of C- and N-terminally His-tagged enzyme in an endogenous PDE yhjH-lacking Escherichia coli strain and in strain BL21(DE3)
commercial synthetic gene PA4781, DNA and amino acid sequence determination and analysis, recombinant overexpression of wild-type and mutant C- and N-terminally His-tagged enzymes in an endogenous PDE yhjH-lacking Escherichia coli strain and in strain BL21(DE3), the REC domain of PA4781 cannot be allosterically activated by phosphorylation in the Escherichia coli background
DNA fragments encoding GGDEF (1-172 aa) and GGDEF-EAL domain (10-426 aa) amplified separately and fused to the GST coding region in pGEX-6p-1, transformation of Escherichia coli BL21. Deletion of Xcc1959 in Xcc strain XC1 or 8004 had no significant effect on virulence factor production in vitro
ectopic expression of 31 of the conserved genes from 20 genomes of Clostridium difficile for comparison of their effect on motility and biofilm formation. Most of the PDEs are active in a Vibrio cholerae model
Flagellar promoter analysis in the bpdA, bpdB, and cgsB mutants using a luminescent reporter system. Expression in Vibrio parahaemolyticus strain LM5984
gene pdeB, gene expression profiling and quantitative PCR in wild-type and mutant cells, expression of GFP-tagged mutant enzymes, expression of N-terminally maltose-binding protein tagged and C-terminally His6 tagged truncated wild-type enzyme and mutant E364A in Escherichia coli strain Tuner(DE3)pLysS
mutated DAA domain cloned in expression vector pLAFR3 (lac promoter) for complementation analysis in a Xanthomonas campestris XC1 mutant lacking ravR. Deletion of ravR results in decreased virulence factor production. RavS and RavR constitute a two-component regulatory system (ravS gene deleted inframe in Xanthomonas campestris strains XC1 and 8004): no effect on bacterial growth in YEB and LB medium, similar phenotype changes compared to Xanthomonas campestris XC1 mutant lacking ravR. RavR is a c-di-GMP phosphodiesterase: fusionated domains GGDEF and EAL (cloned separately into the expression vector pLAFR3 for in trans expression in the mutant deltaravR). RavS/RavR 2-component system regulates diverse functions in Xanthomonas campestris XC1
overexpression is sufficient to induce lateral flagellar gene expression in liquid, decrease biofilm formation, decrease cps gene expression, and suppress the DeltascrABC phenotype
VCA0895 is overexpressed in Vibrio cholerae luxO D47Eel: growth inhibition, deletion of vca0895 from the Vibrio cholerae El genome does not alter biofilm formation (transcriptional activator vps expression)
3.1.4.52 cyclic-guanylate-specific phosphodiesterase
results (
results (
top
