Development and use of a selectable, broad-host-range reporter transposon for identifying environmentally regulated promoters in bacteria.

  title={Development and use of a selectable, broad-host-range reporter transposon for identifying environmentally regulated promoters in bacteria.},
  author={Jennifer K. Spinler and Sheryl L. W. Zajdowicz and Jon C. Haller and Diana Marra Oram and Ronald E. Gill and Randall K. Holmes},
  journal={FEMS microbiology letters},
  volume={291 2},
This report describes the development and use of TnKnXSp, a selectable broad-host-range reporter transposon with a promoterless aphA gene. Insertion of TnKnXSp into the chromosome of a kanamycin-susceptible bacterium confers resistance to kanamycin only if aphA is transcribed from an active promoter adjacent to the insertion site. We designed TnKnXSp as a tool for identifying environmentally regulated promoters in bacteria and developed general methods for initial characterization of any… 
Phage Conversion and the Role of Bacteriophage and Host Functions in Regulation of Diphtheria Toxin Production by Corynebacterium diphtheriae
Corynebacterium diphtheriae is the etiologic agent of diphtheria. Toxinogenic isolates of C. diphtheriae produce diphtheria toxin, a protein that inhibits protein synthesis in susceptible eukaryotic
Corynebacterium diphtheriae Proteome Adaptation to Cell Culture Medium and Serum
Growth and proteome adaptation of Corynebacterium diphtheriae strain ISS3319 were investigated and hint at an increased bacterial pathogenicity when grown in cell culture medium as well as an influence of the growth medium on the cell envelope.
The Genus Corynebacterium


Identification and characterization of three new promoter/operators from Corynebacterium diphtheriae that are regulated by the diphtheria toxin repressor (DtxR) and iron
Three additional DtxR-regulated promoter/operators from C. diphtheriae designated IRP3, IRP4, and IRP5 are identified and characterized and polypeptide products of the predicted sizes were detected in small amounts by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Cloning, sequence, and footprint analysis of two promoter/operators from Corynebacterium diphtheriae that are regulated by the diphtheria toxin repressor (DtxR) and iron
The cloned ORF1 gene encodes a membrane-associated lipoprotein that may serve as the receptor for a ferric-siderophore complex in C. diphtheriae, and nucleotide sequences of the two promoter/operator regions indicated that they are both homologous with the tox operator.
Genetic and Phenotypic Analysis of Acinetobacter baumannii Insertion Derivatives Generated with a Transposome System
DNA hybridization experiments showed that the presence of strain 19606 genes encoding regulatory and attachment or biofilm functions is widespread among other A. baumannii clinical isolates.
Transposon Mutagenesis of the Obligate Intracellular Pathogen Rickettsia prowazekii
A gene that codes for rifampin resistance for expression in R. prowazekii is adapted by incorporating this selection into a transposon mutagenesis system suitable for generating rickettsial gene knockouts, thereby identifying nonessential ricksettsial genes.
Construction of Rhodococcus random mutagenesis libraries using Tn5 transposition complexes.
The absence of efficient transposon systems in Rhodococcus organisms has here been overcome by the use of Tn5-based DNA-protein transposition complexes which can transpose at high efficiency.
Identification of a DtxR-Regulated Operon That Is Essential for Siderophore-Dependent Iron Uptake in Corynebacterium diphtheriae
These findings demonstrated that the irP6 operon in C. diphtheriae encodes a putative ABC transporter, that specific mutant alleles of irp6A, irp4B, and irp5C are associated with defects in corynebactin-dependent iron uptake, and that complementation of these mutant allels restores repression of corynesbact in production under high-iron growth conditions.
Transcription of the Contiguous sigB, dtxR, and galE Genes in Corynebacterium diphtheriae: Evidence for Multiple Transcripts and Regulation by Environmental Factors
These studies provide the first quantitative transcriptional analysis of a gene encoding a DtxR family regulator and give new insights into transcriptional regulation in C. diphtheriae.
Analysis of the Corynebacterium diphtheriae DtxR Regulon: Identification of a Putative Siderophore Synthesis and Transport System That Is Similar to the Yersinia High-Pathogenicity Island-Encoded Yersiniabactin Synthesis and Uptake System
The findings from this study indicate that C. diphtheriae contains at least 18 DtxR binding sites and that D txR may affect the expression of as many as 40 genes.
Iron-dependent regulation of diphtheria toxin and siderophore expression by the cloned Corynebacterium diphtheriae repressor gene dtxR in C. diphtheriae C7 strains.
A regulatory gene (dtxR) responsible for iron-dependent repression of the toxin and siderophore genes in Corynebacterium diphtheriae was cloned and characterized and Expression of the cloned dtxR determinant did not affect the phenotype of C. diphTheriae C7(beta).
Prediction of DtxR regulon: Identification of binding sites and operons controlled by Diphtheria toxin repressor in Corynebacterium diphtheriae
The analysis shows that DtxR could provide a molecular link between Fe+2-induced Fenton's reaction and protection of DNA from oxidative damage and play an important role in host interaction and virulence by regulating the levels of sortase, a potential vaccine candidate and proteins of secretory system.