E. coliGrowth Inhibition by a High Copy Number Derivative of Plasmid pBR322

Valenzuela, Manuel S.; Ikpeazu, Emeka V.; Siddiqui, K. A. I.

doi:10.1006/bbrc.1996.0339

Cited by 20 publications

(14 citation statements)

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“…Interestingly, transformation of vector (pWH1226, a derivative of pBR322 containing the Acinetobacter origin of replication [15]) or vector containing pld (pACJ01) into both wild-type and ACJ2 cells caused a moderate bacterial growth defect when bacteria were grown in plasmid selection medium containing tetracycline. This same phenotype has been observed for other organisms harboring pBR322 derivatives (24,25,40,41). Nonetheless, pACJ01 appeared to fully restore the invasion properties of ACJ2 cells to the levels of the wild type containing vector alone (Fig.…”

Section: Resultssupporting

confidence: 61%

Inactivation of Phospholipase D Diminishes Acinetobacter baumannii Pathogenesis

Hood²,

et al. 2010

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Acinetobacter baumannii is an emerging bacterial pathogen of considerable health care concern. Nonetheless, relatively little is known about the organism's virulence factors or their regulatory networks. Septicemia and ventilator-associated pneumonia are two of the more severe forms of A. baumannii disease. To identify virulence factors that may contribute to these disease processes, genetically diverse A. baumannii clinical isolates were evaluated for the ability to proliferate in human serum. A transposon mutant library was created in a strain background that propagated well in serum and screened for members with decreased serum growth. The results revealed that disruption of A. baumannii phospholipase D (PLD) caused a reduction in the organism's ability to thrive in serum, a deficiency in epithelial cell invasion, and diminished pathogenesis in a murine model of pneumonia. Collectively, these results suggest that PLD is an A. baumannii virulence factor.

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Section: Resultssupporting

confidence: 61%

Inactivation of Phospholipase D Diminishes Acinetobacter baumannii Pathogenesis

Hood²,

et al. 2010

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“…Large plasmids are known burdens for bacterial cells, retarding the lag phase and reducing the overall fitness of cells (47). In addition, a high copy number of a plasmid and the presence of tetracycline resistance gene in a plasmid can inhibit cell growth (48). Thus, the hampered growth of the rpoE475 mutant carrying the complementation plasmid at stressful conditions is not surprising.…”

Section: Discussionmentioning

confidence: 99%

Alternative Sigma Factor σ ^E Has an Important Role in Stress Tolerance of Yersinia pseudotuberculosis IP32953

Palonen

Lindström

Somervuo

et al. 2013

Appl Environ Microbiol

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Yersinia pseudotuberculosis is an important pathogen that probably survives well in the modern food chain. However, little is known about the mechanisms that allow the growth of this pathogen in foods under stress conditions. The expression of rpoE encoding E was defined by quantitative real-time reverse transcription-PCR. Expression of rpoE was induced at 3°C, 37°C, and 42°C, under exposure to 3% NaCl, 3% ethanol, or high and low pH, in relation to its expression at the optimum growth temperature of 28°C of Y. pseudotuberculosis. Mutation of rpoE either impaired or abolished growth under stresses caused by low or high temperature, low pH, and ethanol. In addition, the growth temperature range of the mutant was significantly diminished compared to that of the wild-type strain IP32953. The results were confirmed with complementation of the mutant. Thus, E plays a significant role in the stress tolerance of Y. pseudotuberculosis IP32953 and probably contributes to the survival of this pathogen in the food chain.

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“…The ease of using antibiotic resistance markers for selection has made them widely used in gene manipulations. However, in some situations resistance genes (9, 13) as well as high-copy-number plasmids (49) can interfere with the analysis of studied phenomena. In addition, growth of cells in antibiotic-containing media results in activation of the chromosome-coded multiple antibiotic resistance system (12).…”

Section: Resultsmentioning

confidence: 99%

The IncP plasmid-encoded cell envelope-associated DNA transfer complex increases cell permeability

et al. 1997

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IncP-type plasmids are broad-host-range conjugative plasmids. DNA translocation requires DNA transferreplication functions and additional factors required for mating pair formation (Mpf). The Mpf system is located in the cell membranes and is responsible for DNA transport from the donor to the recipient. The Mpf complex acts as a receptor for IncP-specific phages such as PRD1. In this investigation, we quantify the Mpf complexes on the cell surface by a phage receptor saturation technique. Electrochemical measurements are used to show that the Mpf complex increases cell envelope permeability to lipophilic compounds and ATP. In addition it reduces the ability of the cells to accumulate K ؉ . However, the Mpf complex does not dissipate the membrane voltage. The Mpf complex is rapidly disassembled when intracellular ATP concentration is decreased, as measured by a PRD1 adsorption assay.Plasmids of incompatibility group P (IncP) are large selftransmissible broad-host-range plasmids carrying multiple antibiotic resistance determinants. The prototype IncP plasmid, RP4 (60,099 bp [40]), can replicate in diverse gram-negative organisms. However, the host range identified by conjugative transfer ability is considerably wider than the range of vegetative replication. Suitable shuttle vector constructions carrying the IncP transfer system allow transfer to occur from gramnegative organisms to gram-positive bacteria and to yeast (for recent review see reference 36). Evolutionary and functional relationships of the IncP DNA transfer system to other systems include Ti/Ri plasmids that direct T-DNA transfer from agrobacteria to plant cells (45). These properties make RP4 a suitable model for studying bacterial conjugation (16,17,24,25,38).Functions responsible for the conjugative transfer of RP4 are encoded in two distinct regions on the plasmid known as Tra1 and Tra2 (Fig. 1). Essential transfer functions and the Tra1 and Tra2 core regions were recently identified and evaluated by genetic analyses (5, 29, 51). Transfer functions are divided into two groups: the DNA transfer and replication (Dtr) system and the mating pair formation (Mpf) apparatus, involved in bringing the donor and the recipient cells into intimate contact during conjugation. Dtr genes map exclusively in Tra1. This region consists of three operons and includes the origin of transfer (oriT), which is an intergenic region between the leader and the relaxase operons. The Dtr functions participate in relaxosome formation, the initiation complex of the transfer-replication process (37,39,50). Transfer of the DNA is thought to occur via TraI-piloted single-stranded intermediates that may be coated by the TraC protein (42, 52). The DNA-TraC complex is thought to be transported through a channel or pore at the mating bridge between the donor and the recipient cells. Formation of this channel or pore depends on gene products of the Mpf system. Mpf genes

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E. coliGrowth Inhibition by a High Copy Number Derivative of Plasmid pBR322

Cited by 20 publications

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Inactivation of Phospholipase D Diminishes Acinetobacter baumannii Pathogenesis

Inactivation of Phospholipase D Diminishes Acinetobacter baumannii Pathogenesis

Alternative Sigma Factor σ ^E Has an Important Role in Stress Tolerance of Yersinia pseudotuberculosis IP32953

The IncP plasmid-encoded cell envelope-associated DNA transfer complex increases cell permeability

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E. coliGrowth Inhibition by a High Copy Number Derivative of Plasmid pBR322

Cited by 20 publications

References 0 publications

Inactivation of Phospholipase D Diminishes Acinetobacter baumannii Pathogenesis

Inactivation of Phospholipase D Diminishes Acinetobacter baumannii Pathogenesis

Alternative Sigma Factor σ E Has an Important Role in Stress Tolerance of Yersinia pseudotuberculosis IP32953

The IncP plasmid-encoded cell envelope-associated DNA transfer complex increases cell permeability

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Alternative Sigma Factor σ ^E Has an Important Role in Stress Tolerance of Yersinia pseudotuberculosis IP32953