scholarly journals Stable RK2-Derived Cloning Vectors for the Analysis of Gene Expression and Gene Function in Gram-Negative Bacteria

2001 ◽  
Vol 14 (3) ◽  
pp. 426-430 ◽  
Author(s):  
Bruno Dombrecht ◽  
Jos Vanderleyden ◽  
Jan Michiels
Keyword(s):  
Gene Expression ◽  
Gene Function ◽  
Plasmid Stability ◽  
Gram Negative Bacteria ◽  
Cloning Vectors ◽  
Gram Negative ◽  
Gusa Gene ◽  
Vector Sequences ◽  
And Function ◽  
Multiple Cloning Sites

The construction of several stable RK2-derived cloning vectors for the analysis of gene expression and function in gram-negative bacteria is reported. Plasmid stability is conferred by the RK2 par locus or by insertion of the spsAB or spsCD symbiotic plasmid stability loci from pNGR234a of Rhizobium sp. NGR234. The vectors carry multiple cloning sites with protection against read-through transcriptional activity of vector sequences. Vector derivatives with the constitutive nptII promoter or a promoter-less gusA gene are suitable for the study of gene function or regulation in bacteria.

scholarly journals Construction of a Broad-Host-Range Tn7-Based Vector for Single-Copy PBAD-Controlled Gene Expression in Gram-Negative Bacteria

2012 ◽  
Vol 79 (2) ◽  
pp. 718-721 ◽  
Author(s):  
F. Heath Damron ◽  
Elizabeth S. McKenney ◽  
Herbert P. Schweizer ◽  
Joanna B. Goldberg
Keyword(s):  
Gene Expression ◽  
Pseudomonas Aeruginosa ◽  
Host Range ◽  
Single Copy ◽  
Broad Host Range ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Content Type ◽  
Delivery Vector ◽  
Inducible Gene

ABSTRACTWe describe a mini-Tn7-based broad-host-range expression cassette for arabinose-inducible gene expression from the PBADpromoter. This delivery vector, pTJ1, can integrate a single copy of a gene into the chromosome of Gram-negative bacteria for diverse genetic applications, of which several are discussed, usingPseudomonas aeruginosaas the model host.

scholarly journals Central Role of Toll-Like Receptor 4 Signaling and Host Defense in Experimental Pneumonia Caused by Gram-Negative Bacteria

2005 ◽  
Vol 73 (1) ◽  
pp. 532-545 ◽  
Author(s):  
Jill R. Schurr ◽  
Erana Young ◽  
Pat Byrne ◽  
Chad Steele ◽  
Judd E. Shellito ◽  
...  
Keyword(s):  
Gene Expression ◽  
Adaptor Protein ◽  
Mouse Strains ◽  
Gram Negative Bacteria ◽  
Toll Like Receptor ◽  
Toll Like Receptor 4 ◽  
Tlr4 Signaling ◽  
Gram Negative ◽  
Experimental Pneumonia

ABSTRACT Toll-like receptor 4 (TLR4) has been identified as a receptor for lipopolysaccharide. However, the precise role of TLR4 in regulating gene expression in response to an infection caused by gram-negative bacteria has not been fully elucidated. The role of TLR4 signaling in coordinating gene expression was assessed by gene expression profiling in lung tissue in a mouse model of experimental pneumonia with a low-dose infection of Klebsiella pneumoniae. We analyzed four mouse strains: C57BL/6 mice, which are resistant to bacterial dissemination; 129/SvJ mice, which are susceptible; C3H/HeJ mice, which are susceptible and have defective TLR4 signaling; and their respective control strain, C3H/HeN (intermediate resistance). At 4 h after infection, C57BL/6 and C3H/HeN mice demonstrated the greatest number of genes, with 67 shared induced genes which were TLR4 dependent and highly associated with the resistance phenotype. These genes included cytokine and chemokine genes required for neutrophil activation or recruitment, growth factor receptors, MyD88 (a critical adaptor protein for TLR signaling), and adhesion molecules. TLR4 signaling accounted for over 74% of the gene expression in the C3H background. These data suggest that early TLR4 signaling controls the vast majority of gene expression in the lung in response to an infection caused by gram-negative bacteria and that this subsequent gene expression determines survival of the host.

Direct selection cloning vectors adapted to the genetic analysis of Gram-negative bacteria and their plasmids

Gene ◽  
1998 ◽  
Vol 207 (1) ◽  
pp. 87-92 ◽  
Author(s):  
Philippe Gabant ◽  
Cédric Y Szpirer ◽  
Martine Couturier ◽  
Michel Faelen
Keyword(s):  
Genetic Analysis ◽  
Direct Selection ◽  
Gram Negative Bacteria ◽  
Cloning Vectors ◽  
Gram Negative

scholarly journals Plasposons: Modular Self-Cloning Minitransposon Derivatives for Rapid Genetic Analysis of Gram-Negative Bacterial Genomes

1998 ◽  
Vol 64 (7) ◽  
pp. 2710-2715 ◽  
Author(s):  
Jonathan J. Dennis ◽  
Gerben J. Zylstra
Keyword(s):  
Broad Host Range ◽  
Gram Negative Bacteria ◽  
Sequence Specificity ◽  
Origin Of Replication ◽  
Bacterial Genomes ◽  
Gram Negative ◽  
Dna Sequence Specificity ◽  
Target Dna ◽  
Versatile Tool ◽  
Multiple Cloning Sites

A series of modular mini-transposon derivatives which permit the rapid cloning and mapping of the DNA flanking the minitransposon’s site of insertion has been developed. The basic plasposon, named TnMod, consists of the Tn5 inverted repeats, a conditional origin of replication, rare restriction endonuclease multiple cloning sites, and exchangeable antibiotic resistance cassettes. The broad host range and low target DNA sequence specificity of the Tn5 transposase, in combination with the flexibility afforded by the modular arrangement of TnMod, result in a versatile tool for the mapping of insertional mutations and the rapid recovery of clones from gram-negative bacteria.

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