Improved gfp and inaZ broad-host-range promoter-probe vectors.
TLDR
A new set of broad-host-range promoter-probe vectors has been constructed that contains the pVS1 and p15a replicons and confers resistance to either gentamicin or kanamycin and is bound by transcriptional terminators to permit the insertion of strong promoters and to insulate the cassette from external transcription enabling the detection of weak or moderate promoters.Abstract:
A new set of broad-host-range promoter-probe vectors has been constructed. One subset contains the pVS1 and p15a replicons and confers resistance to either gentamicin or kanamycin. The other set contains the broad-host-range replicon from pBBR1 and confers resistance to kanamycin, tetracycline, ampicillin, or spectinomycin/streptomycin. Both plasmid sets are highly stable and are maintained without selection for more than 30 generations in several bacterial taxa. Each plasmid contains a promoter-probe cassette that consists of a multicloning site, containing several unique restriction sites, and gfp or inaZ as a reporter gene. The cassette is bound by transcriptional terminators to permit the insertion of strong promoters and to insulate the cassette from external transcription enabling the detection of weak or moderate promoters. The vector suite was augmented with derivatives of the kanamycin-resistant gfp promoter-probe plasmids that encode Gfp variants with different half-life times.read more
Citations
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Environmentally Controlled Invasion of Cancer Cells by Engineered Bacteria
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The Standard European Vector Architecture (SEVA): a coherent platform for the analysis and deployment of complex prokaryotic phenotypes
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TL;DR: Using in vivo fluorescence measurements, immunolabeling, and quantitative gene expression analysis, it is demonstrated that S. oneidensis MR-1 nanowires are extensions of the outer membrane and periplasm that include the multiheme cytochromes responsible for EET, rather than pilin-based structures as previously thought.
References
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Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans.
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Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes.
Michael E. Kovach,Philip H. Elzer,D. Steven Hill,Gregory T. Robertson,Michael A. Farris,R. Martin Roop,Kenneth M. Peterson +6 more
TL;DR: In this paper, four new antibiotic-resistant derivatives of the broad-host-range (bhr) cloning vector pBBR1MCS have been constructed, which are relatively small (< 5.3 kb), possess an extended multiple cloning site (MCS), and allow direct selection of recombinant plasmid molecules in Escherichia coli via disruption of the LacZ alpha peptide.
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Analysis of gene control signals by DNA fusion and cloning in Escherichia coli
TL;DR: Plasmid cloning vectors that enable insertion of DNA fragments between the inducible ara (arabinose) promoter and the lac (lactose) structural genes have been constructed and used for the detection and analysis of signals that control gene transcription.
Journal ArticleDOI
Molecular basis of bacterial outer membrane permeability.
Hiroshi Nikaido,Martti Vaara +1 more
TL;DR: It is becoming increasingly clear that the outer membrane is very important in the physiology of gram-negative bacteria in making them resistant to host defense factors such as lysozyme, P-lysin, and various leukocyte proteins.
Journal ArticleDOI
In vitro insertional mutagenesis with a selectable DNA fragment
Pierre Prentki,Henry M. Krisch +1 more
TL;DR: A new method for in vitro insertional mutagenesis of genes cloned in Escherichia coli that makes use of the omega fragment, a 2.0-kb DNA segment consisting of an antibiotic resistance gene flanked by short inverted repeats carrying transcription and translation termination signals and synthetic polylinkers.