Pseudomonas fluorescens HK44 is a lux-based bioluminescent bioreporter capable of emitting light upon exposure to naphthalene, salicylate, and other substituted analogs. The bacterium was inoculated ...into intermediate-scale field lysimeters and population dynamics were monitored with time. Two methods were used to enumerate cell numbers in soil: a standard selective plating technique with colony hybridization verification and a modified lux-based most-probable-number (lux-MPN) assay based on the detection of bioluminescence. The lux-MPN assay was developed and evaluated as a possible supplement or replacement for the labor-intensive and time-consuming selective plating assay. Comparisons between selective plate counts and lux-MPN population estimates showed similar trends over the 2-year study, except that lux-MPN estimates were consistently less than selective plate counts. Verification of P. fluorescens HK44 genotype through colony hybridization techniques revealed that selective plating was actually overestimating HK44 populations and that lux-MPN values were more closely approximating true HK44 cell densities, except within the first few weeks after inoculation, when lux-MPN estimates underrepresented population densities. Thus, utilizing bioluminescence as a population monitoring tool for lux-based microorganisms was shown to be more effective and precise than standard selective plating techniques, and provided an accurate ecological analysis of P. fluorescens HK44 population dynamics over an extended period.
A bioluminescent reporter strain, Acinetobacter sp. DF4-8, was constructed for the detection of phenol by inserting a mopR-like promoter upstream of the Vibrio fischeri bioluminescent luxCDABE gene ...cassette in a modified mini-Tn5 construct. When introduced into the chromosome of Acinetobacter sp. DF4, the bioreporter produced a sensitive bioluminescent response to phenol at concentrations ranging from 2.5 to 100 ppm. This response was linear (R(2)=0.986) in the range from 20 to 90 ppm. A significant bioluminescent response was also recorded when strain DF4-8 was incubated with slurries from aged, phenol-contaminated soil.
We report an integrated CMOS microluminometer for the detection of low-level bioluminescence in whole cell biosensing applications. This microluminometer is the microelectronic portion of the ...bioluminescent bioreporter integrated circuit (BBIC). This device uses the n-well/p-substrate junction of a standard bulk CMOS IC process to form the integrated photodetector. This photodetector uses a distributed electrode configuration that minimizes detector noise. Signal processing is accomplished with a current-to-frequency converter circuit that forms the causal portion of the matched filter for dc luminescence in wide-band white noise. Measurements show that luminescence can be detected from as few as 4×10
5 cells/ml.
Pseudomonasfluorescens HK44 is a lux-based bioluminescent bioreporter capable of selective luminescence in the presence of naphthalene and/or salicylic acid intermediate of its metabolism. We ...attempted to induce bioluminescence (BL) in this strain with 72 compounds, viz. substituted naphthalenes, naphthalene-like compounds (e.g., quinoline), substituted salicylic acids, salicylic acid-like compounds (e.g., 2-anthranilic acid), oligocyclic aromates, and intermediates of naphthalene metabolism to better discriminate response specificity. From them, 42 induced BL significantly lower as compared to naphthalene, three (viz. isoquinoline, o-cresol, and salicylamide) induced BL significantly greater than naphthalene, and 27 yielded no bioluminescent response whatsoever. Strain HK44 is therefore not prone to extensive false-positive signaling and can serve as a fairly specific indicator organism for naphthalene bioavailability. At elevated concentrations, 41 compounds inhibited BL. Thus, the inclusion of constitutive bioreporter controls as indicators of sample toxicity is vital to successful biosensing application.
The composition and nature of high-Cr minerals in lithic clasts from the carbonatites of the Veseloe occurrence, northern Transbaikalia, were considered. In order to determine their source, the ...Cr-bearing phases were compared with chromite, magnetite, and rutile from ultrabasic rocks, mantle xenoliths, and eclogites. It was suggested that the xenoclasts studied were formed at great depths, whereas the carbonatites were directly derived from the mantle rather than formed by the crustal differentiation of a silicate-carbonate melt.PUBLICATION ABSTRACT
We report an integrated CMOS microluminometer optimized for the detection of low-level bioluminescence as part of the bioluminescent bioreporter integrated circuit (BBIC). This microluminometer ...improves on previous devices through careful management of the sub-femtoampere currents, both signal and leakage, that flow in the front-end processing circuitry. In particular, the photodiode is operated with a reverse bias of only a few mV, requiring special attention to the reset circuitry of the current-to-frequency converter (CFC) that forms the front-end circuit. We report a sub-femtoampere leakage current and a minimum detectable signal (MDS) of 0.15
fA (1510
s integration time) using a room temperature 1.47
mm
2 CMOS photodiode. This microluminometer can detect luminescence from as few as 5000 fully induced
Pseudomonas fluorescens 5RL bacterial cells.
This paper presents the results of a study of the Vesely carbonatite occurrence in a new carbonatite-bearing area of northern Transbaikalia. The REE patterns, oxygen and carbon isotope compositions ...of carbonates and magnetites, and strontium isotope composition of dolomite allow us to classify the rocks as carbonatites. This study focused on minerals from xenoliths that were identified as mantle-derived on the basis of structure and mineral composition. The compositions of Cr-bearing chlorite, phengite, magnetite, rutile, ilmenite, and titanite are reported. Aggregates of closely intergrown magnetite and rutile occurring in the xenoliths resemble exsolution structures. A possible origin of these aggregates is discussed.PUBLICATION ABSTRACT
Metamorphism of carbonatite is exemplified in the Vesely occurrence. According to available data, the age of the carbonatite is 596 ± 3.5Ma, whereas metamorphism is dated at 550 ± 14 Ma. The rocks at ...the Vesely occurrence were metamorphosed under conditions of greenschist facies (epidote-muscovite-chlorite subfacies) under elevated pressure. Microthermometry of fluid inclusions in minerals indicates that the temperature of metamorphism is 377−450°C and the pressure estimated from phengite geobarometer is 6−8 kbar. The low-grade metamorphism led to the partial recrystallization of carbonates and apatite with removal of trace elements. This process resulted in a change of the oxygen isotopic composition of the studied minerals. Metamorphism was accompanied by formation of talc, phengite, chlorite, quartz, tremolite-actinolite, and anthophyllite, which are not typical of carbonatite. The data obtained show that the metamorphism exerted an effect on the mineralogical, isotopic, geochemical, and technological properties of the carbonatite. The effect of metamorphism should be taken into account in determination of the nature of ore mineralization and estimation of ore quality and perspective of the occurrence.
Tocotrienols are the primary form of vitamin E in seeds of most monocot plants, including cereals such as rice and wheat. As potent antioxidants, tocotrienols contribute to the nutritive value of ...cereal grains in human and livestock diets. cDNAs encoding homogentisic acid geranylgeranyl transferase (HGGT), which catalyzes the committed step of tocotrienol biosynthesis, were isolated from barley, wheat and rice seeds. Transgenic expression of the barley HGGT in Arabidopsis thaliana leaves resulted in accumulation of tocotrienols, which were absent from leaves of nontransformed plants, and a 10- to 15-fold increase in total vitamin E antioxidants (tocotrienols plus tocopherols). Overexpression of the barley HGGT in corn seeds resulted in an increase in tocotrienol and tocopherol content of as much as six-fold. These results provide insight into the genetic basis for tocotrienol biosynthesis in plants and demonstrate the ability to enhance the antioxidant content of crops by introduction of an enzyme that redirects metabolic flux.
The vertical movement of a genetically-engineered bacterium released into a sub-surface soil contained within a lysimeter was investigated. The strain used was
Pseudomonas fluorescens HK44, a
...lux-based microbe capable of bioremediating specific polyaromatic hydrocarbons (PAHs). Strain HK44 was inoculated into a 4
m deep by 2.5
m diameter soil lysimeter outfitted with a hydraulic assembly to control groundwater levels. After 230 days, the lysimeter was subjected to a water table manipulation that resulted in significant transport of HK44 cells to depths 60
cm below the original inoculation zone within 20 days. A lysimeter not exposed to water level fluctuations exhibited no significant vertical transport. These results demonstrate the potential distribution of genetically engineered microorganisms in subsurface soil due to groundwater effects.
