Use of <i>lux</i> genes in applied biochemistry

Hill, Philip J.; Stewart, Gordon S. A. B.

doi:10.1002/bio.1170090315

Cited by 20 publications

(13 citation statements)

References 28 publications

(16 reference statements)

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“…Bioluminescence should be infrequent enough among typical feed microorganisms that background microbial growth would not interfere with the bioassay. Bioluminescent E. coli strains cells can be constructed by genetically transferring genes responsible for phenotypic lumination from marine-dwelling bacteria to terrestrial species that naturally do not contain these genes (Baker et al 1992;Hill and Stewart 1994). Consequently a bioluminescent signal can not only be coupled to bacterial growth response to accurately measure levels of the respective nutrient limiting bacterial growth but is 10-fold more sensitive than OD .…”

Section: Introductionmentioning

confidence: 99%

CONSTRUCTION AND GROWTH KINETICS OF A BIOLUMINESCENT METHIONINE AUXOTROPH ESCHERICHIA COLI STRAIN FOR POTENTIAL USE IN A METHIONINE BIOASSAY

Froelich

Díaz

Ricke

2002

Rapid Methods Automation Mic

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Methionine is one of the essential and first limiting amino acids in animal nutrition. In this study, an Escherichia coli methionine auxotroph bacterial strain that exhibits a linear growth response to methionine concentrations was transformed with a plasmid containing genes encoding ampicillin resistance and bioluminescence in order to develop a microbiological technique for methionine quantitation. Transformants were selected based on antibiotic resistance and plasmid containing candidates were confirmed by restriction enzyme digestion and gel electrophoresis. To confirm the bioluminescent phenotype, video imaging of the strain using long exposure photography yielded colonies exhibiting bioluminescence. The strain was also tested in the presence of ampicillin supplemented media with increasing methionine concentrations and growth response (measured as optical density, OD), growth rates and methionine affinities were compared before and after transformation. Although the transformed E. coli methionine auxotroph exhibited somewhat different growth kinetic responses than the nontransformed strain, the standard curves used for estimating methionine concentrations were not different. Based on the results in this study the transformed bioluminescent strain could be used as an OD‐based assay if bioluminescence equipment and materials are not available.

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Section: Introductionmentioning

confidence: 99%

CONSTRUCTION AND GROWTH KINETICS OF A BIOLUMINESCENT METHIONINE AUXOTROPH ESCHERICHIA COLI STRAIN FOR POTENTIAL USE IN A METHIONINE BIOASSAY

Froelich

Díaz

Ricke

2002

Rapid Methods Automation Mic

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“…The cloning of structural genes encoding bacterial luciferase from Vibrio harveyi [45] and from Vibrio fischerii [46], firefly luciferase [47], and Renilla luciferase from Renilla reniformis [48] allows luciferase-mediated luminescence imaging [49,50] to be widely used in biomedical research. All luciferases have a substrate requirement, e.g.…”

Section: Localization Of Light Emission In Animals Injected With Labementioning

confidence: 99%

Optical imaging: bacteria, viruses, and mammalian cells encoding light-emitting proteins reveal the locations of primary tumors and metastases in animals

Timiryasova

Zhang

et al. 2003

Anal Bioanal Chem

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Early detection of tumors and their metastases is crucial for the prognosis of cancer treatment. Traditionally, tumor detection is achieved by various methods, including magnetic resonance imaging and computerized tomography. With the recent cloning, cellular expression, and real-time imaging of light-emitting proteins, such as Renilla luciferase (Ruc), bacterial luciferase (Lux), firefly luciferase (Luc), green fluorescent protein (GFP), or Ruc-GFP fusion protein, significant efforts have been focused on using these marker proteins for tumor detection. It has also been demonstrated that certain bacteria, viruses, and mammalian cells (BVMC), when administered systemically, are able to gain entry and replicate selectively in tumors. In addition, many tissue/tumor specific promoters have been cloned which allow transgene expression specifically in tumor tissues. Therefore, when light-emitting protein encoded BVMC are injected systemically into rodents, tumor-specific marker gene expression is achieved and is detected in real time based on light emission. Consequently, the locations of primary tumors and previously unknown metastases in animals are revealed in vivo. In the future it will likely be feasible to use engineered light-emitting BVMC as probes for tumor detection and as gene-delivery vehicles in vivo for cancer therapy.

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“…In E. coli , bioluminescence does not occur and must be acquired via genetic modifications [118,119]. Luminescence is accomplished by the introduction of the luxAB genes via plasmid or chromosomal insertion.…”

Section: Detection Modes For Methionine Microbial Biosensorsmentioning

confidence: 99%

Potential for Development of an Escherichia coli—Based Biosensor for Assessing Bioavailable Methionine: A Review

Chalova

Froelich

Ricke

2010

Sensors

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Methionine is an essential amino acid for animals and is typically considered one of the first limiting amino acids in animal feed formulations. Methionine deficiency or excess in animal diets can lead to sub-optimal animal performance and increased environmental pollution, which necessitates its accurate quantification and proper dosage in animal rations. Animal bioassays are the current industry standard to quantify methionine bioavailability. However, animal-based assays are not only time consuming, but expensive and are becoming more scrutinized by governmental regulations. In addition, a variety of artifacts can hinder the variability and time efficacy of these assays. Microbiological assays, which are based on a microbial response to external supplementation of a particular nutrient such as methionine, appear to be attractive potential alternatives to the already established standards. They are rapid and inexpensive in vitro assays which are characterized with relatively accurate and consistent estimation of digestible methionine in feeds and feed ingredients. The current review discusses the potential to develop Escherichia coli-based microbial biosensors for methionine bioavailability quantification. Methionine biosynthesis and regulation pathways are overviewed in relation to genetic manipulation required for the generation of a respective methionine auxotroph that could be practical for a routine bioassay. A prospective utilization of Escherichia coli methionine biosensor would allow for inexpensive and rapid methionine quantification and ultimately enable timely assessment of nutritional profiles of feedstuffs.

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Use of lux genes in applied biochemistry

Cited by 20 publications

References 28 publications

CONSTRUCTION AND GROWTH KINETICS OF A BIOLUMINESCENT METHIONINE AUXOTROPH ESCHERICHIA COLI STRAIN FOR POTENTIAL USE IN A METHIONINE BIOASSAY

CONSTRUCTION AND GROWTH KINETICS OF A BIOLUMINESCENT METHIONINE AUXOTROPH ESCHERICHIA COLI STRAIN FOR POTENTIAL USE IN A METHIONINE BIOASSAY

Optical imaging: bacteria, viruses, and mammalian cells encoding light-emitting proteins reveal the locations of primary tumors and metastases in animals

Potential for Development of an Escherichia coli—Based Biosensor for Assessing Bioavailable Methionine: A Review

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