Generation of Vibrio anguillarum Ghost by Coexpression of PhiX 174 Lysis E gene and Staphylococcal Nuclease A Gene

Kwon, Se Ryun; Kang, Yue Jai; Lee, Dong Jin; Lee, Eun Hye; Nam, Yoon Kwon; Kim, Sung Koo; Kim, Ki Hong

doi:10.1007/s12033-009-9147-y

Cited by 32 publications

(13 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Plasmid pRKkPR-cI-E-SNA bearing the bacteriophage PhiX174 lysis E gene was introduced into normal E. coli S17-1 cells to produce E. coli ghosts (42). Although these E. coli cells are preyed upon by B. bacteriovorus 109J, their ghosts were not recognized as prey when added to a fresh culture of AP B. bacteriovorus 109J.…”

Section: Resultsmentioning

confidence: 99%

Cell-cycle progress in obligate predatory bacteria is dependent upon sequential sensing of prey recognition and prey quality cues

Rotem

Pasternak

Shimoni

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Predators feed on prey to acquire the nutrients necessary to sustain their survival, growth, and replication. In Bdellovibrio bacteriovorus, an obligate predator of Gram-negative bacteria, cell growth and replication are tied to a shift from a motile, free-living phase of search and attack to a sessile, intracellular phase of growth and replication during which a single prey cell is consumed. Engagement and sustenance of growth are achieved through the sensing of two unidentified prey-derived cues. We developed a novel ex vivo cultivation system for B. bacteriovorus composed of prey ghost cells that are recognized and invaded by the predator. By manipulating their content, we demonstrated that an early cue is located in the prey envelope and a late cue is found within the prey soluble fraction. These spatially and temporally separated cues elicit discrete and combinatory regulatory effects on gene transcription. Together, they delimit a poorly characterized transitory phase between the attack phase and the growth phase, during which the bdelloplast (the invaded prey cell) is constructed. This transitory phase constitutes a checkpoint in which the late cue presumably acts as a determinant of the prey's nutritional value before the predator commits. These regulatory adaptations to a unique bacterial lifestyle have not been reported previously.microbial ecology | bacterial physiology | bacterial cell cycle | predatory bacteria | Bdellovibrio bacteriovorus

show abstract

Section: Resultsmentioning

confidence: 99%

Cell-cycle progress in obligate predatory bacteria is dependent upon sequential sensing of prey recognition and prey quality cues

Rotem

Pasternak

Shimoni

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…More than 80 serotypes of V. parahaemolyticus have been described worldwide, based on the O and K antigenic properties [22]. To date, some BGs have been produced from other Vibrio species ( V. cholera , V. anguillarum , V. vulnificus ) by using the lysis E -gene system [4,5,23,24], but no BGs have been reported from V. parahaemolyticus . In the present study, we determined the MIC of various chemicals (six acids and one alkali) on V. parahaemolyticus cells and characterized their efficiencies to produce V. parahaemolyticus ghosts (VPGs).…”

Section: Introductionmentioning

confidence: 99%

Characterization of Chemically-Induced Bacterial Ghosts (BGs) Using Sodium Hydroxide-Induced Vibrio parahaemolyticus Ghosts (VPGs)

Park

Vinod

et al. 2016

IJMS

View full text Add to dashboard Cite

Acellular bacterial ghosts (BGs) are empty non-living bacterial cell envelopes, commonly generated by controlled expression of the cloned lysis gene E of bacteriophage PhiX174. In this study, Vibrio parahaemolyticus ghosts (VPGs) were generated by chemically-induced lysis and the method is based on minimum inhibitory concentration (MIC) of sodium hydroxide (NaOH), acetic acid, boric acid, citric acid, maleic acid, hydrochloric acid, and sulfuric acid. The MIC values of the respective chemicals were 3.125, 6.25, <50.0, 25.0, 6.25, 1.56, and 0.781 mg/mL. Except for boric acid, the lysis efficiency reached more than 99.99% at 5 min after treatment of all chemicals. Among those chemicals, NaOH-induced VPGs appeared completely DNA-free, which was confirmed by quantitative real-time PCR. Besides, lipopolysaccharides (LPS) extracted from the NaOH-induced VPGs showed no distinctive band on SDS-PAGE gel after silver staining. On the other hand, LPS extracted from wild-type bacterial cells, as well as the organic acids-induced VPGs showed triple major bands and LPS extracted from the inorganic acids-induced VPGs showed double bands. It suggests that some surface structures in LPS of the NaOH-induced VPGs may be lost, weakened, or modified by the MIC of NaOH. Nevertheless, Limulus amoebocyte lysate assay revealed that there is no significant difference in endotoxic activity between the NaOH-induced VPGs and wild-type bacterial cells. Macrophages exposed to the NaOH-induced VPGs at 0.5 × 106 CFU/mL showed cell viability of 97.9%, however, the MIC of NaOH did not reduce the cytotoxic effect of wild-type bacterial cells. Like Escherichia coli LPS, the NaOH-induced VPGs are an excellent activator of pro-inflammatory cytokines (IL-1β and iNOS), anti-inflammatory cytokine (IL-10), and dual activities (IL-6) in the stimulated macrophage cells. On the other hand, the induction of TNF-α mRNA was remarkable in the macrophages exposed with wild-type cells. Scanning electron microscopy showed the formation of trans-membrane lysis tunnel structures in the NaOH-induced VPGs. SDS-PAGE and agarose gel electrophoresis also confirmed that cytoplasmic proteins and genomic DNA released from the VPGs to culture medium through the lysis tunnel structures. Taken together, all these data indicate that the NaOH-induced VPGs show the potency of a safe, economical, and effective inactivated bacterial vaccine candidate.

show abstract

“…Even highly sensitive and fragile structures such as pili can be well protected by this technology (Eko et al, 2000). The potential usefulness of bacterial ghosts as vaccine candidates against a wide variety of pathogenic Gram-negative bacteria (Ra et al, 2009), such as EHEC O157 Cai et al, 2010), EPEC E2348/69 (Liu et al, 2012), Vibrio anguillarum (Kwon et al, 2009), Aeromonas hydrophila and Edwardsiella tarda (Lee et al, 2008;Wang and Lu, 2009), has recently been widely reported. However, production and characterization of bacterial ghosts for a vaccine candidate against an intracellular bacterial pathogen, such as Brucella, have not been reported.…”

Section: Introductionmentioning

confidence: 99%

Immune responses and protection induced by Brucella suis S2 bacterial ghosts in mice

Liu

Sun

et al. 2015

Veterinary Immunology and Immunopathology

View full text Add to dashboard Cite

Generation of Vibrio anguillarum Ghost by Coexpression of PhiX 174 Lysis E gene and Staphylococcal Nuclease A Gene

Cited by 32 publications

References 22 publications

Cell-cycle progress in obligate predatory bacteria is dependent upon sequential sensing of prey recognition and prey quality cues

Cell-cycle progress in obligate predatory bacteria is dependent upon sequential sensing of prey recognition and prey quality cues

Characterization of Chemically-Induced Bacterial Ghosts (BGs) Using Sodium Hydroxide-Induced Vibrio parahaemolyticus Ghosts (VPGs)

Immune responses and protection induced by Brucella suis S2 bacterial ghosts in mice

Contact Info

Product

Resources

About