Unknown,Transcriptomics,Genomics,Proteomics

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Vibrio campbellii bioluminescence and its effect on gene expression and virulence

ABSTRACT: The selective advantage of bioluminescence in bacterial cells that do not form symbiotic relationships with aquatic animals is still not known. Some evidence suggests that bioluminescence plays a role in DNA repair by a photoreactivation process (Czyz 2000) and that non-bioluminescent strains are less virulent than their bioluminescent isogenic counterparts (Ruwandeepika 2010). All hypotheses to date suggest bioluminescence associated or mediated changes in gene expression, yet the evidence for this does not exist. In this study, we generated an in-frame luxAB deletion mutant (the two contiguous genes that encode for bacterial luciferase) and compared its mid-log phase gene expression profile with that of the wild type spontaneous streptomycin resistant (STR) V. campbellii BAA-1116 parental strain from which it was derived. Both mid-log phase transcriptomes were elucidated using custom designed whole genome microarrays (520694F, Affymetrix) to determine the effect luciferase has on V. campbellii gene expression. The virulence phenotypes of both strains were also subsequently tested in Artemia franciscana challenge experiments. Three biological replicates of the wild type (STR) and luxAB deletion mutant of V. campbellii BAA-1116 were grown to mid-log phase (15 h, 200 rpm, 30M-BM-0C, in 25 mL autoinducer bioassay medium) and total RNA was extracted from 1.0E+9 cells. Messenger RNA was isolated from the total RNA extracts treated with DNase, labeled with biotin, fragmented and hybridized to V. campbellii BAA-1116 whole genome microarrays (520694F, Affymetrix).

ORGANISM(S): Vibrio campbellii

SUBMITTER: Baochuan Lin

PROVIDER: E-GEOD-34741 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Introduction: Tuberculosis (TB) is a serious human disease caused by the bacterium Mycobacterium tuberculosis (Mtb). One third of the worldâs population is estimated to be latently infected with Mtb. Exact mechanisms and properties of latent TB infection (LTBI) are not fully elucidated. In essence, the pathogen can enter a dormant or latent state characterized by limited growth and metabolism, resulting in absence of clinical symptoms in the host, and most importantly by increased phenotypic tolerance to the commonly used medications, thereby allowing indefinite persistence in the human body. This persistence is the main reason why the current treatment of new-onset pulmonary TB is very long, consisting of a six months therapy of four antibiotics (rifampicin, isoniazid, pyrazinamide, and ethambutol for the first two months, and only rifampicin and isoniazid for the last four months). Current state of the art: To fight TB globally and more efficiently, it is essential to shorten the treatment for TB with new drugs that are efficient also against LTBI. To facilitate discovery of such drugs, in vitro models for LTBI can be used for high throughput screening of chemical libraries. Current in vitro models such as nutrient starvation (Betts et al. 2002), nutrient depletion (Hampshire et al. 2004; Rifat, Bishai, and Karakousis 2009), progressive hypoxia (Wayne and Hayes 1996), nitric oxide treatment (Martin I. Voskuil et al. 2003) and multiple stress (Deb et al. 2009) mimic the dormant state of Mtb, but the technical settings and equipment required for these models obstruct high throughput applications. Proposed model: The streptomycin (STR)-starved 18b model (SS18b) is a simple and drug discovery efficient Mtb latency model successfully applicable to both in vitro and in vivo settings (Zhang et al. 2012). It is based on the Mtb strain 18b, which is a STR-dependent mutant that enters a viable but nonreplicating state in the absence of STR (Hashimoto 1955). Despite the successful model, we know very little about 18b. How well does SS18b mimic the bacterial response to the complex host-pathogen interactions underlying LTBI, and how does the SS18b response compare to that of other dormancy models are still open questions. In this work we analyse the complete genome of 18b and describe the transcriptomic response of 18b to STR depletion. 12 samples were analyzed. Four biological replicates were sampled during the exponential growth phase, in presence of streptomycin. Four biological replicates derived after 2 weeks of streptomycin depletion, and two biological replicates derive after 4 weeks of streptomycin depletion. Two replicates derive from the resubmission of streptomycin after four weeks of depletion.

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Vibrio campbellii bioluminescence and its effect on gene expression and virulence

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