Robert J Mitchell

Ulsan National Institute of Science and Technology | UNIST · School of Life Sciences

One beguiling alternative to antibiotics for treating multi-drug resistant infections are Bdellovibrio-and-like-organisms (BALOs), predatory bacteria known to attack human pathogens. Consequently, in this study, the responses from four cell lines (three human and one mouse) were characterized during an exposure to different predatory bacteria, Bdel...

Bdellovibrio bacteriovorus HD100 is a predatory bacterium that attacks many Gram-negative human pathogens. A serious drawback of this strain, however, is its ineffectiveness against Gram-positive strains, such as the human pathogen Staphylococcus aureus. Here we demonstrate that the extracellular proteases produced by a host-independent B. bacterio...

Micropatterning of bacteria using aqueous two phase system (ATPS) enables the localized culture and formation of physically separated bacterial communities on human epithelial cell sheets. This method was used to compare the effects of Escherichia coli strain MG1655 and an isogenic invasive counterpart that expresses the invasin (inv) gene from Yer...

Plant-based cellulose presents the best source of renewable sugars for biofuel production. However, the lignin associated with plant cellulose presents a hurdle as hydrolysis of this component leads to the production of inhibitory compounds, such as ferulic acid. The impacts of ferulic acid, a phenolic compound commonly found in lignin hydrolysates...

Identified as a newly described species from a biocrust in Svalbard, Norway (78° 54′ 8.27″ N 12° 01′ 20.34″ E), isolate PAP01 T has different characteristics from any known predatory bacteria. The isolate was vibrio-shaped strain that employed flagellar motility. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the isolate clust...

In a survey of the International Space Station (ISS), the most common pathogenic bacterium identified in samples from the air, water and surfaces was Staphylococcus aureus. While growth under microgravity is known to cause physiological changes in microbial pathogens, including shifts in antibacterial sensitivity, its impact on S. aureus is not wel...

Biofilms are a major concern within the food industry since they have the potential to reduce productivity in situ (within the field), impact food stability and storage, and cause downstream food poisoning. Within this review, predatory bacteria as potential biofilm control and eradication agents are discussed, with a particular emphasis on the int...

ABSTRACT While diverse antibacterials are available in nature, each possesses their own strengths and limitations. One such antibacterial is colicins, proteinaceous toxins that are produced by strains of E. coli to subvert the growth or viability of other E. coli strains. Similarly, predatory bacteria, of which Bdellovibrio bacteriovorus is well-kn...

With the growing threat of antibiotic resistance, researchers around the globe are seeking alternatives to stem bacterial pathogenesis. One such alternative is bacteriocins, proteins produced by bacterial species to inhibit the growth and viability of related bacterial species. With their diverse mechanisms, which include pore formation and nucleas...

Biosensors are utilized in several different fields, including medicine, food, and the environment; in this review, we examine recent developments in biosensors for healthcare. These involve three distinct types of biosensor: biosensors for in vitro diagnosis with blood, saliva, or urine samples; continuous monitoring biosensors (CMBs); and wearabl...

Bdellovibrio and like organisms (BALOs) are a unique bacterial group that live by predating on other bacteria, consuming them from within to grow and replicate before the progeny come out to complete the life cycle. The mechanisms by which these predators recognize their prey and differentiate them from nonprey bacteria, however, are still not clea...

Bdellovibrio and like organisms are predatory bacteria that are capable of attacking, killing, and consuming many bacterial pathogens, including multidrug-resistant strains. These qualities have led to them being labeled as “living antibiotics.” Research work with these remarkable strains, however, has been hampered by long growth times needed to q...

As interest in natural biogenic silica nanoparticles (NPs) from rice husks grows, it is important to know how their preparation (i.e., combustion, acid leaching, or alkali extraction) impacts their biocompatibility. Acid leaching and alkali extraction both generated highly pure silica NPs (>99.1% SiO2vs 93.1% with combustion). However, toxicity tes...

The antibacterial activities of prodigiosin are well-established but, as a hydrophobic molecule, the mechanisms used to introduce it to susceptible microbes has never been studied. We found here, in contrast to violacein, another hydrophobic antibiotic that can be transferred using membrane vesicles (MVs), prodigiosin is also carried from Serratia...

As mankind evaluates moving toward permanently inhabiting outer space and other planetary bodies, alternatives to antibiotic that can effectively control drug-resistant pathogens are needed. The activity of one such alternative, Bdellovibrio bacteriovorus HD100, was explored here, and was found to be as active or better in simulated microgravity (S...

In this review, we discuss violacein and prodigiosin, two chromogenic bacterial secondary metabolites that have diverse biological activities. Although both compounds were “discovered” more than seven decades ago, interest into their biological applications has grown in the last two decades, particularly driven by their antimicrobial and anticancer...

Bdellovibrio bacteriovorus 109J is a predatory bacterium which lives by predating on other Gram-negative bacteria to obtain the nutrients it needs for replication and survival. Here, we evaluated the effects two classes of bacterial signaling molecules (acyl homoserine lactones (AHLs) and diffusible signaling factor (DSF)) have on B. bacteriovorus...

This study demonstrates the impact outer membrane permeability has on the power densities generated by E. coli-based microbial fuel cells with neutral red as the mediator, and how increasing the permeability improves the current generation. Experiments performed with several lipopolysaccharide (LPS) mutants (ΔwaaC, ΔwaaF and ΔwaaG) of E. coli BW251...

Recent years have witnessed increased interest in systems that are capable of supporting multistep chemical processes without the need for manual handling of intermediates. These systems have been based either on collections of batch reactors¹ or on flow-chemistry designs2–4, both of which require considerable engineering effort to set up and contr...

Mutations that shorten the lipopolysaccharide (LPS) in Escherichia coli were found to significantly increase the number of transformants after electroporation. The loss of the LPS outer core increased the number of transformants with plasmid pAmCyan (3.3 kb) from 5.0 × 10⁵ colony-forming units (CFU)/μg in the wild-type E. coli BW25113 to 3.3 × 10⁷...

‘Everything is everywhere, but the environment selects’

Maternal behaviors benefit the survival of young, contributing directly to the mother’s reproductive fitness. An extreme form of this is seen in matriphagy, when a mother performs the ultimate sacrifice and offers her body as a meal for her young. Whether matriphagy offers only a single energy-rich meal or another possible benefit to the young is u...

Bdellovibrio-and-like organisms (BALOs) are a small group of bacteria that actively predate on other Gram-negative bacterial species. Although viewed mostly in a positive light, such as their potential use as living antibiotics to reduce pathogenic strain populations, several studies have also highlighted the need to control their activities, such...

This work is a collection of articles that discuss microbial predation from a variety of perspectives. It provides the readers a concise resource describing factors that are critical for several different predatory microbes, including Myxobacterium spp. and Bdellovibrio­-and-like organisms (BALOs), including the mechanisms involved, ecological cond...

This study describes Chromobacterium violaceum’s use of extracellular membrane vesicles (MVs) to both solubilize and transport violacein to other microorganisms. Violacein is a hydrophobic bisindole with known antibiotic activities against other microorganisms. Characterization of the MVs found they carried more violacein than protein (1.37±0.19‐fo...

We evaluated the toxicity of surfactants against different predatory bacteria. Tests with Bdellovibrio bacteriovorus HD100 and SDS, an anionic surfactant, showed the predator was very sensitive; 0.02% SDS completely killed the predatory population (7-log loss; < 10 PFU/ml remaining) both when free-swimming or within the bdelloplast, i.e., intraperi...

B. bacteriovorus HD100 is a highly motile predatory bacterium that consumes other Gram‐negative bacteria for its sustenance. Here we describe the impacts the media viscosity has both on the motility of predator and its attack rates. Experiments performed in polyethylene glycol (PEG) solutions, a linear polymer, found a viscosity of 10 mPa‐s (5% PEG...

Abstract In this study we show Yersinia pseudotuberculosis secretes membrane vesicles (MVs) that contain different proteins and virulence factors depending on the strain. Although MVs from Y. pseudotuberculosis YPIII and ATCC 29833 had many proteins in common (68.8% of all the proteins identified), those located in the outer membrane fraction diffe...

Bdellovibrio bacteriovorus HD100 is a predatory bacterium which lives by invading the periplasm of Gram-negative bacteria and consuming them from within. Although B. bacteriovorus HD100 attacks only Gram-negative bacterial strains, our work here shows attack-phase predatory cells also benefit from interacting with Gram-positive biofilms. Using Stap...

Predation of Chromobacterium piscinae by Bdellovibrio bacteriovorus HD100 was inhibited in dilute nutrient broth (DNB) but not in HEPES. Experiments showed that the effector responsible was present in the medium, as cell-free supernatants retained the ability to inhibit predation, and that the effector was not toxic to B. bacteriovorus. Violacein,...

We evaluated the bactericidal activity of Bdellovibrio bacteriovorus, strain HD100, within blood sera against bacterial strains commonly associated with bacteremic infections, including E. coli, Klebsiella pneumoniae and Salmonella enterica. Tests show that B. bacteriovorus HD100 is not susceptible to serum complement or its bactericidal activity....

The prevalence of Staphylococcus aureus worldwide as a nosocomial infectious agent is recognized but the reason behind the spread of this bacterium has remained elusive. Here, we hypothesized that the communication of S. aureus might benefit from it blocking other bacteria from establishing themselves on the surface. This was found to be the case f...

Bdellovibrio bacteriovorus is a predatory bacterium which lives by invading the periplasm of gram-negative bacteria and consuming them from within. This predator was thought to be dependent upon prey for nutrients since it lacks genes encoding for critical enzymes involved in amino acid biosynthesis. This study, however, found that planktonic attac...

Violacein is a bisindole antibiotic that is effective against Gram-positive bacteria while the bacterial predator, Bdellovibrio bacteriovorus HD100, predates on Gram-negative strains. In this study, we evaluated the use of both together against multidrug resistant pathogens. The two antibacterial agents did not antagonize the activity of the other....

Aims: Evaluate the use of polyethyleneimines (PEIs) as membrane permeabilizers to improve the responses and sensitivity of a bacterial bioreporter strain to viologens. Methods and results: The responses from E. coli str. EBS, i.e., E. coli BW25113 carrying plasmid pSDS, when exposed to five different viologens was characterized, as were the toxi...

Electronic-cigarettes (e-cigarette) are widely used as an alternative to traditional cigarettes but their safety is not well established. Herein, we demonstrate and validate an analytical method to discriminate the deleterious effects of e-cigarette refills (e-juice) and soluble e-juice aerosol (SEA) by employing stress-specific bioluminescent reco...

Forty-nine pigments were extracted from the collections of 106 pigment producing bacteria from the plant rhizosphere soil. Antibacterial activity test was performed in the subjects of the extracted pigments with plant pathogenic bacteria including Xanthomonas axonopodis and Xanthomonas campestris, and with plant pathogenic fungi including Botrytis...

As a field within the energy research sector, bioenergy is continuously expanding. Although much has been achieved and the yields of both ethanol and butanol have been improved, many avenues of research to further increase these yields still remain. This review covers current research related with transcriptomics and the application of this high-th...

A violacein-producing bacterial strain was isolated and identified as a relative of Duganella violaceinigra YIM 31327 based upon phylogenetic analyses using the 16S rRNA, gyrB and vioA gene sequences and a fatty acid methyl ester (FAME) analysis. This new strain was designated D. violaceinigra str. NI28. Although these two strains appear related ba...

This paper describes an optimization of planetary mill pretreatment and saccharification processes for improving biosugar production. Pitch pine ( Pinus rigida ) wood sawdust waste was used as biomass feedstock and the process parameters optimized in this study were the buffering media, the milling time, the enzyme quantity, and the incubation time...

The effects of three single variable parameters on the glucose yields were investigated before measuring the combinational effects of experimental conditions. Figure S1 shows the effects each of the individual variables had on the glucose yields. As increasing the value of each variable, the productions of glucose were increased and reached to satu...

A periodic jet of carbon dioxide (CO 2) aerosols is a very quick and effective mechanical technique to remove biofilms from various substrate surfaces. However, the impact of the aerosols on the viability of bacteria during treatment has never been evaluated. In this study, the effects of high-speed CO 2 aerosols, a mixture of solid and gaseous CO...

Violacein-producing bacteria, with their striking purple hues, have undoubtedly piqued the curiosity of scientists since their first discovery. The bisindole violacein is formed by the condensation of two tryptophan molecules through the action of five proteins. The genes required for its production, vioABCDE , and the regulatory mechanisms employe...

This study evaluated the co-application of bacterial predation by Bdellovibrio bacteriovorus and either alum coagulation or powdered activated carbon adsorption to reduce fouling caused by Escherichia coli rich feed solutions in dead-end microfiltration tests. The flux increased when the samples were predated upon or treated with 100 ppm alum or PA...

We describe a novel method which makes use of polymer-based aqueous two-phase systems to pattern bacterial communities inside Petri dishes. This method allows us to culture submillimeter-sized bacterial communities in spatially distinct spots while maintaining a degree of chemical connectedness to each other through the aqueous phase. Given suffici...

Bdellovibrio bacteriovorus is a predatory bacterium which attacks a wide range of Gram-negative bacterial pathogens and is proposed to be a potential living antibiotic. In this study, we evaluated the effects of indole, a bacterial signaling molecule commonly produced within the gut, on the predatory ability of B. bacteriovorus HD100. Indole signif...

This study assessed the dynamics of predation by Bdellovibrio bacteriovorus HD 100. Predation tests with two different bioluminescent strains of Escherichia coli, one expressing a heat-labile bacterial luciferase and the other a heat-stable form, showed near identical losses from both, indicating that protein expression and stability are not respon...

RT-qPCR data demonstrated that the yhcN gene was highly expressed when Escherichia coli was exposed to 500 ppm of ferulic acid (9.8-fold), vanillin (7.3-fold) or furfural (2.2-fold). Consequently, an E. coli yhcN::lux bioreporter strain (DMY1) was constructed. This strain is very responsive to furfural and 5-hydroxymethylfurfural, showing a maximum...

Membrane biofouling is one of the major obstacles limiting membrane applications in water treatment. In this study, Bdellovibrio bacteriovorus HD 100, a Gram-negative predatory bacterium, was evaluated as a novel way to mitigate membrane biofouling and its subsequent performance decline. Dead-end microfiltration (MF) tests were carried out on Esche...

This study evaluated the use of planetary and attrition milling as pretreatment processes for lignocellulosic biomass using rice straw. Planetary milling reduced the rice straw crystallinity from 0.48 to 0.11. Since the samples could be milled and enzymatically treated using the same media, loss of the biomass due to washing was effectively elimina...

Effect of initial rOD of P. sp on the damage caused to the underlying MCF 10a cells layer. The central photo shows the P. sp DSM 50906 communities made using four different initial rOD values (5, 0.5, 0.05, and 0.005) all were spotted as 0.3 µl droplets on the MCF 10a monolayer in the same plate. The analysis was done after 24 h of incubation at 37...

Comparing ATPS culturing with the ordinary culturing of bacteria on polystyrene surface. E. coli MG1655 was spotted in plain 35 mm Petri dishes (with no epithelial cells cultured), at different rOD values using both ordinary culturing and the ATPS technique. For the former, the bacteria were resuspended into ordinary DMEM/F12 medium at the required...

The microscopical appearance of the epithelial layer underneath E. coli and S. boydii ATPS derived spots. E. coli MG1655 and S. boydii KACC 10792 ATPS derived communities were not harmful to the underlying MCF 10a cells. This is clear from the integrity and well being of the epithelial cell monolayer underneath each of these two bacterial spots. Im...

The macroscopical appearance of ATPS derived E. coli, S. boydii, and P. sp bacterial communities on MCF 10a monolayer. The three strains were spotted as 0.3 µl droplets on a single MCF 10a monolayer at an initial rOD of 0.5 in DMEM/F12 medium. E. coli MG1655 and S. boydii KACC 10792 formed clearly visible bacterial communities after 24 h of incubat...

Comparing three different ATPS formulations for possible use for patterning bacteria on epithelial cells. MCF 10a cells were stained with calcein AM and EthD-1 (Live/Dead) stain and observed under epifluorescence microscope after 24 h of incubation with the PEG rich phase of each of the three formulations and the ordinary DMEM/F12 medium alone. A)...

Background Currently, the most promising microorganism used for the bio-production of butyric acid is Clostridium tyrobutyricum ATCC 25755T; however, it is unable to use sucrose as a sole carbon source. Consequently, a newly isolated strain, Bacillus sp. SGP1, that was found to produce a levansucrase enzyme, which hydrolyzes sucrose into fructose a...

Bacterial bioreporters are limited in their abilities to detect large polar molecules due to their membrane selectivity. In this study, the activity of serum complement was used to bypass this undesired selectivity. Initially, the serum complement activity was assessed using the responses of a bacterial bioreporter harboring a recA::luxCDABE transc...

Growth of B. sp. SGP1 in RCM adjusted to different pH values. This experiment was done in serum bottles without previous purging with argon and the OD600 was measured after 6 h of growth at 37°C in a shaking incubator.

Effect of temperature on sucrase activity of the supernatant of B. sp. SGP1. B. sp. SGP1 was grown for 22 h in RCM supplemented with sucrose, then 50 ul of the supernatant was taken and added to 950 μl of sucrose solution (0.5 M in phosphate buffer, at pH 6.6, and different temperature) and then the liberated glucose was measured as described in th...

Effect of temperature on butyric acid production by the co-culture. This experiment was done in serum bottles using RCM supplemented with sucrose.

Neighbor-joining tree based on nearly complete 16S rRNA gene sequences showing the relationships between the strain SGP1 and its related strains. The percentage numbers at the nodes are the levels of bootstrap support based on neighbor-joining analyses of 1000 resampled data sets. Marinococcus halophilus DSM 20408T (X90835) was used as the out grou...

This paper describes a system to study how small physical perturbations can affect bacterial community behavior in unexpected ways through modulation of diffusion and convective transport of chemical communication molecules and resources. A culture environment that mimics the chemically open characteristic of natural bacterial habitats but with use...

A bioluminescent Escherichia coli bioreporter strain to detect hydrolysate related phenolics was developed by cloning the aaeXAB promoter from E. coli upstream of the luxCDABE genes. E. coli str. DH5α carrying this plasmid (pDMA3) was responsive to sub-inhibitory concentrations of plant hydrolysate-related phenolics, such as ferulic and vanillic ac...

This study evaluated predation with Bdellovibrio bacteriovorous and CO(2) aerosol spraying to remove fluorescent Escherichia coli biofilms from silicon chips. Initial tests found that 7.5×10(5) viable E. coli cells were dispersed into the surrounding environment during aerosol treatment. The total number dispersed per test decreased to only 16 for...

We describe patterning of bacterial biofilms using polymer-based aqueous two-phase system (ATPS) microprinting protocols. The fully aqueous but selectively bacteria-partitioning nature of the ATPS allows spatially distinct localization of suspensions of bacteria such as Pseudomonas aeruginosa and Escherichia coli with high precision. The ATPS patte...

Lignin hydrolysates contain many different chemical species, including ferulic acid, coumaric acid, vanillic acid, vanillin, syringaldehyde and furfural. From the perspective of biofuels, these compounds are problematic and can cause downstream loss of product if not removed prior to beginning the fermentative process. In contrast, a search for the...

In this paper, the different applications of butyric acid and its current and future production status are highlighted, with a particular emphasis on the biofuels industry. As such, this paper discusses different issues regarding butyric acid fermentations and provides suggestions for future improvements and their approaches.

Aberrations in the growth and transcriptome of Escherichia coli str. BL21(DE3) were determined when exposed to varying concentrations of ferulic acid (0.25-1 g/L), an aromatic carboxylic acid identified within lignin-cellulose hydrolysate samples. The expression of several individual genes (aaeA, aaeB, inaA and marA) was significantly induced, i.e....

Bdellovibrio bacteriovorus is a predatory bacterium which attacks and consumes other bacterial strains, including the well known pathogens E. coli O157 : H7, Salmonella typhimurium and Helicobacter pylori. This remarkable activity has been the focus of research for nearly five decades, with exciting practical applications to medical, agriculture an...

This study presents a simple protocol to measure 2 promoter activities within a single culture when using both Lux and firefly luciferase (FF-Luc) reporters. To demonstrate this, 2 E. coli strains were constructed using 2 compatible plasmids, one harboring a katG::luc fusion gene and the other either a fabA::lux or grpE::lux fusion gene. To differe...

We present a microfabricated concentrator array device that makes it possible to quantify the predation rate of Bdellovibrio bacteriovorus, a predatory microbe, toward its prey, Escherichia coli str. MG1655. The device can accumulate both prey and predator microbes sequentially within a series of concentrator arrays using the motility of the microb...

Real-time quantitative PCR analyses of Escherichia coli str. BL21(DE3) exposed to 0.5 g/L ferulic and coumaric acid showed that the inaA gene was significantly induced (7.7- and 3.6-fold higher, respectively). Consequently, a transcriptional fusion of the inaA promoter with the luxCDABE operon was constructed and characterized with several compound...

Inter-cellular communication via diffusible small molecules is a defining character not only of multicellular forms of life but also of single-celled organisms. A large number of bacterial genes are regulated by the change of chemical milieu mediated by the local population density of its own species or others. The cell density-dependent "autoinduc...

Using an aqueous two-phase system comprised of dextran and polyethylene glycol, this article describes the stable spatial patterning of sub-microlitre droplets of bacterial suspensions. Microdroplets of different types of bacterial populations are positioned and maintained adjacent to each other without significant dispersion even though the bacter...

Starting with the first publication of lacZ gene fusion in 1980, reporter genes have just entered their fourth decade. Initial studies relied on the simple fusion of a promoter or gene with a particular reporter gene of interest. Such constructs were then used to determine the promoter activity under specific conditions or within a given cell or or...

To answer the need for a non-food based source of fermentable sugar, researchers have focused on woody plants, which can consist of 70% or more sugar, in the form of hemicellulose and cellulose. During hydrolysis of the wood, however, the lignin fraction is also hydrolyzed, producing aromatic acids and aldehydes, such as ferulic acid, coumaric acid...

The ultimate goal in the production of biofuels is to produce fuels identical or similar to petroleum-derived transportation fuels more efficiently and in commercial quantities. Synthetic biologists have been engineering microbes to synthesize biofuels, such as butanol and fatty acid- or isoprenoid-based fuels, which are nearly identical to gasolin...

The effects of the hydraulic retention time (HRT=8, 10, 12 or 16.7 h) and glucose concentration (30, 40 or 50 g/L) on the production of hydrogen and butyrate by an immobilized Clostridium tyrobutyricum culture, grown under continuous culturing conditions, were evaluated. With 30 g/L glucose, the higher HRTs tested led to greater butyrate concentrat...

The toxicity of several compounds commonly found in lignin hydrolysates were evaluated individually by monitoring the production of acetone, butanol and ethanol (ABE) from Clostridium beijerinckii. The compounds selected for analysis included coumaric acid, syringaldehyde, furfural, ferulic acid and glucuronic acid. Assays with C. beijerinckii were...

The recombinant bioluminescent bacterium, DNT5, containing a nagR-nagAa::luxCDABE fusion, was tested in a multi-channel continuous monitoring system to evaluate its ability to detect benzoic acid derivatives. Seven chemicals, benzoic acid, salicylic acid, 2,5-dihydroxy benzoic acid, 3,5-dihydroxy benzoic acid, benzene, naphthalene and phenol, were...

Twelve recombinant bioluminescent bacteria have been immobilized within the wells of a 96-well plate using two different matrices--agar and sol-gel. All 12 strains were immobilized within individual wells of the plates and the sensitivity of the strains and the stability of the responses were determined for select chemicals. Each strain was exposed...

A kan'::luxCDABE fusion strain that was both highly bioluminescent and responsive to benzoic acid was constructed by transforming E. coli strain W3110 with the plasmid pUCDK, which was constructed by digesting and removing the 7-kb KpnI fragment from the promoterless luxCDABE plasmid pUCD615. Experiments using buffered media showed that this induct...

A kan’::luxCDABE fusion strain that was both highly bioluminescent and responsive to benzoic acid was constructed by transforming E. coli strain W3110 with the plasmid pUCDK, which was constructed by digesting and removing the 7-kb KpnI fragment from the promoterless luxCDABE plasmid pUCD615. Experiments using buffered media showed that this induct...

In this study, a cell-based array technology that uses recombinant bioluminescent bacteria to detect and classify environmental toxicity has been implemented to develop two biosensor arrays, i.e., a chip and a plate array. Twenty recombinant bioluminescent bacteria, having different promoters fused with the bacterial lux genes, were immobilized wit...

The NagR protein is a response regulatory protein found in the bacterium Ralstonia sp. U2 that is involved in sensing for salicylic acid and the subsequent induction of the operon just upstream of its gene. The genes encoded for in this operon are involved in the degradation of salicylic acid. Escherichia coli strain RFM443 carrying a fusion of the...

The enhancement of the multi-channel continuous toxicity monitoring system developed previously was studied. To achieve better and more stable results from the system, the use of thermo-lux fusion strains that express the luxCDABE genes from Xenorhabdus luminescens was evaluated. A total of six recombinant Escherichia coli strains with the promoter...

A recombinant bioluminescent Escherichia coli DH5 strain, EBJM1 (pqi-5::luxCDABE), was constructed in this study by fusing the promoter for the pqi-5 gene, which is a member of the soxRS regulon but with an unknown function, to the Vibrio fisheri luxCDABE operon within plasmid pUCD615. The soxRS-dependent activation of the pqi-5 promoter was seen w...

A comparison of promoter fusions with the luxCDABE genes from Vibrio fischeri and Photorhabdus luminescens was made using promoters from several genes (katG, sodA, and pqi-5) of E. coli that are responsive to oxidative damage. The respective characteristics, such as the basal and maximum bioluminescence and the relative bioluminescence, were compar...

Using the genes for the green fluorescence protein and Xenorhabdus luminescens luciferase operon and the promoters for the recA and katG genes, two stress-responsive Escherichia coli biosensor strains have been constructed that can individually or concurrently respond to oxidative and genotoxic conditions. Strain DUO-1 carries the pRGDK1 plasmid, w...

A two-plasmid dual reporter Escherichia coli biosensor was developed using the genes for bacterial bioluminescence and a mutant of the green fluorescent protein, GFPuv4. To achieve this, the two plasmids, which were derivatives of pBR322 and pACYC184, had compatible origins of replication and different antibiotic selection markers: ampicillin and t...