University of Tennessee Research Foundation

Ut-battelle, Inc.

Other

Eastman Chemical Company

United States of America As Represented by the Secretary of the Air Force

International Technology Corporation

Gary S Sayler

Steven Ripp

Michael Leon Simpson

Bruce Michael Applegate

Michael J Paulus

Alice C Layton

Curtis A Lajoie

James T Fleming

Christine Jo Kelly

Michael P Conway

Mounir Laroussi

Daniel M Close

Stacy S Patterson

Battle B Glascock

Rakesh K Gupta

Raymond Stapleton

James W Blackburn

Cells, including stem cells, comprising an autobioluminescent phenotype, wherein the cells emit a luminescent signal in the absence of an exogenous luminescent stimulator, are provided. The luminescent signal may be constitutive, inducible, repressible, or tissue-specific. The cells express a synthetically engineered bacterial luciferase (lux) cassette, i.e., the luxCDABEfrp gene cassette. The cells may comprise luxA, luxB, luxC, luxD, luxE, and flavin reductase. The cells may each express a combined expression level of luxC, luxD, luxE, and flavin reductase that is from ten to forty times greater than a combined expression level of luxA and luxB. Further, methods of making and using the cells comprising an autobioluminescent phenotype are disclosed herein.

Lux expression in cells and methods of use

An integrated microluminometer includes an integrated circuit chip having at least one n-well/p-substrate junction photodetector for converting light received into a photocurrent, and a detector on the chip for processing the photocurrent. A distributed electrode configuration including a plurality of spaced apart electrodes disposed on an active region of the photodetector is preferably used to raise efficiency.

Microluminometer chip and method to measure bioluminescence

The luxA, B, C, D, and E genes fromhave been introduced intobioluminescent yeast cells.

Lux expression in eukaryotic cells

Monolithic bioelectronic devices for the detection of ammonia includes a microorganism that metabolizes ammonia and which harbors a lux gene fused with a heterologous promoter gene stably incorporated into the chromosome of the microorganism and an Optical Application Specific Integrated Circuit (OASIC). The microorganism is generally a bacterium.

Bioluminescent bioreporter integrated circuit devices and methods for detecting ammonia

Bioelectronic devices for the detection of estrogen include a collection of eukaryotic cells which harbor a recombinant lux gene from a high temperature microorganism wherein the gene is operably linked with a heterologous promoter gene. A detectable light-emitting lux gene product is expressed in the presence of the estrogen and detected by the device.

Bioluminescent bioreporter integrated circuit devices and methods for detecting estrogen

A method for cell-based combinatorial logic includes the steps of providing at least one genetically engineered cell, the genetically engineered cell having at least one transcriptional unit. The transcriptional unit includes a gene and a promoter, wherein application of a stimulus to the promoter results in the expression of a gene product. An energetic or chemical stimulus is applied to activate the promoter, wherein the detection of an output signal corresponds to the presence of a gene product. The cell can include a plurality of transcriptional units configured to form logic gates. The logic gates of a plurality of cells can be operably interconnected by release of output signals, such as chemical stimuli.

Methods for cell-based combinatorial logic

Disclosed are monolithic bioelectronic devices comprising a bioreporter and an OASIC. These bioluminescent bioreporter integrated circuit are useful in detecting substances such as pollutants, explosives, and heavy-metals residing in inhospitable areas such as groundwater, industrial process vessels, and battlefields. Also disclosed are methods and apparatus for detection of particular analytes, including ammonia and estrogen compounds.

Bioluminescent bioreporter integrated circuit detection methods

Disclosed are bioluminescent bioreporter integrated circuit devices that detect selected analytes in fluids when implanted in the body of an animal. The device comprises a bioreporter that has been genetically engineered to contain a nucleic acid segment that comprises a cis-activating response element that is responsive to the selected substance operably linked to a gene encoding a bioluminescent reporter polypeptide. In preferred embodiments, the target analyte is glucose, glucagons, or insulin. Exposure of the bioreporter to the target substance causes the response element to up-regulate the nucleic acid sequence encoding the reporter polypeptide to produce a luminescent response that is detected and quantitated. In illustrative embodiments, the bioreporter device is encapsulated on an integrated circuit that is capable of detecting the emitted light, processing the resultant signal, and then remotely reporting the results. Also disclosed are controlled drug delivery systems capable of being directly or indirectly controlled by the detection device that provide drugs such as insulin to the animal in reponse to the amount of target analyte present in the body fluids.

In vivo biosensor apparatus and method of use

Disclosed are methods and devices for detection of bacteria based on recognition and infection of one or more selected strains of bacteria with bacteriophage genetically modified to cause production of an inducer molecule in the bacterium following phage infection. The inducer molecule is released from the infected bacterium and is detected by genetically modified bacterial bioreporter cells designed to emit bioluminescence upon stimulation by the inducer. Autoamplification of the bioluminescent signal permits detection of low levels of bacteria without sample enrichment. Also disclosed are methods of detection for select bacteria, and kits for detection of select bacteria based on the described technology.

Bioluminescent biosensor device

Voltage is applied to conducting loops wrapped around the outside of a non-conducting chamber (e.g., a glass tube) to generate a capacitively coupled discharge plasma inside the chamber. In one embodiment, a seed gas is injected into the chamber through an inlet in an otherwise closed end of the chamber, while the other end is open to the ambient atmosphere. In such an embodiment, the seed gas is used to ignite the plasma in air at essentially atmospheric pressure. The present invention has different applications, including, but not limited to, (a) passivating toxic or polluting gases that are injected into the chamber along with the seed gas and (b) treating materials placed within a second chamber that is connected to the open end of the plasma-generating chamber such that active species migrate into the second chamber to interact with the materials placed therein.

Electrodeless discharge at atmospheric pressure

The invention provides two new zoogloeal strains, mz1t and mz2t. The invention provides an isolated nucleic acid consisting of the nucleic acid of SEQ ID NO:1. Examples of nucleic acids of mz1t include SEQ ID NO:2 and SEQ ID NO:3. An example of a nucleic acid of mz2t is SEQ ID NO:4. The invention also provides an isolated nucleic acid consisting of the nucleic acid of SEQ ID NO:5. A method of detecting the presence of zoogloeal clusters in a wastewater sample is provided, comprising: a) contacting RNA from a sample of the wastewater with a nucleic acid comprising the nucleic acid of SEQ ID Nos:1,2,3,4 or 5 under conditions that permit specific hybridization; and b) detecting the presence of hybridization, the presence of hybridization indicating the presence of zoogloeal clusters. A new Hyphomicrobium spp. strain, designated M3, is provided herein. The invention provides novel nucleic acids of a Hyphomicrobium sp. M3. For example SEQ ID NO:6 and SEQ ID NO:7 rDNAs of M3. SEQ ID Nos:8, 9, 10, 11, 12, 13 and 14 are presumptive Hyphomicrobium spp. sequences. SEQ ID NO:15 is a consensus sequence from 6m3 and 3m3 (Hyphomicrobium sp. m3 isolated from sludge. A method of detecting the presence of a Hyphomicrobium species in a wastewater sample as described above, using a nucleic acid comprising the nucleic acid of SEQ ID NO:17 is also provided. The nucleic acids of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12 SEQ ID NO:13, SEQ ID NO:14 and SEQ ID NO:15 can also be utilized in whole or as fragments in the detection method for Hyphomicrobium spp.

Zoogloeal and hyphomicrobium spp. nucleic acids

Disclosed are monolithic bioelectronic devices comprising a bioreporter and an OASIC. These bioluminescent bioreporter integrated circuit are useful in detecting substances such as pollutants, explosives, and heavy-metals residing in inhospitable areas such as groundwater, industrial process vessels, and battlefields. Also disclosed are methods and apparatus for environmental pollutant detection, oil exploration, drug discovery, industrial process control, and hazardous chemical monitoring.

Bioluminescent bioreporter integrated circuit

The present invention provides a reporter bacterium, comprising a bacterium that occurs naturally in a biological sludge and that contains a nucleic acid that encodes a reporter protein not found in the naturally occurring bacterium. The nucleic acid can encode a bioluminescent reporter protein. A method and apparatus are also provided for detecting the presence of toxicity in a wastewater treatment influent stream, comprising contacting the influent with a reporter bacterium of the present invention; monitoring the expression of the reporter protein by the reporter bacterium; and correlating a reduction in the expression of the reporter protein with the presence of toxicity. In an additional embodiment, the present invention provides a method and apparatus for determining the effect of a known dose of an oxidative phosphorylation decoupling agent on biological wastewater sludge, comprising contacting a sample of the sludge containing a known dose of the decoupling agent and a control sample, with a reporter bacterium of the present invention; comparing the signal output by the reporter bacteria in the sludge sample containing the decoupling agent with the signal output by the reporter bacteria in the sludge sample not containing the decoupling agent; and correlating a decrease in signal output in the decoupled sample as compared to the signal output in the control sample with an action of the decoupling agent, whereby the effect of the known dose of the decoupling agent is determined.

Bioluminescent reporter bacterium

The differential display (DD) technique, widely used previously for eukaryotic gene discovery, is optimized to detect differential mRNA transcription (an expressed gene) from both pure culture and soil derived bacterial RNA. A model system using toluene induction of TodC1 in Pseudomonas putida F1 is used to optimize the procedure. Once optimized, an arbitrary primer for the RT step in conjunction with the same arbitrary primer and a Shine-Dalgarno (SD) primer for the PCR reaction is used to detect the expressed gene. The invention thus provides a method for discovery and acquisition of novel genes from environmental microbial communities that avoids the traditional steps and inherent bias due to the culturing of environmental isolates.

Isolation of expressed genes in microorganisms

New strains of microorganisms which posses the dual capabilities for growth on surfactants and degradation of polychlorinated biphenyls are used in combination with soil washing for PCB bioremediation. Soil to be treated is washed with a surfactant to solubilize the generally hydrophobic contaminants. The surfactant solution is then treated in a bioreactor with the microorganisms. As the surfactant is degraded, the residual desolubilized contaminants are adsorbed onto an inert substrate, which is removed from the effluent and can be recycled to the bioreactor.

Growing community of inventors

Blaine, TN, United States of America

Gary S Sayler