E. coli Active Beta Glucuronidase (DAG-P2704)

E. coli Active Beta Glucuronidase (full length), recombinant protein from E. coli

Product Overview
Active E. coli Beta Glucuronidase (GUS) full length protein
Nature
Recombinant
Tag/Conjugate
Unconjugated
Cellular Localization
Cytoplasmic
Bio-activity
Activity: >10,000,000 units/g protein. This enzyme does not hydrolyze alpha-glucuronides or beta-glucosides. This preparation is essentially free of sulfatase activity.
Procedure
1mM EDTA
Format
Lyophilised
Buffer
Preservative: None Constituents: Polyethylene glycol (as stabilizer), 10mM Potassium phosphate, 1mM EDTA, 1mM DTT
Preservative
None
Storage
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze/thaw cycles.
Reconstitution
Reconstitute with 75mM phosphate buffer, pH 6.8, to give a 5mg/ml solution. This solution will be clear to slightly hazy but is still active regardless.
Introduction
Escherichia coli; commonly abbreviated E. coli) is a gram-negative, facultatively anaerobic, rod-shaped bacterium of the genus Escherichia that is commonly found in the lower intestine of warm-blooded organisms (endotherms). Most E. coli strains are harml
Antigen Description
Reporter genes are widely used for studying the expression of foreign genes in transformed plants tissues. Using appropriate promoter-reporter gene constructs, this technique allows an independent verification of the transformed status of tissues growing on media containing selective antibiotics or herbicides. In addition, it serves as a principal means to follow gene transfer and monitor genetic transformation of plant species. Encoded by the E. coli GUS gene (also referred to as uidA), GUS protein is a hydrolase that catalyses the cleavage of a variety of beta-glucuronide derivatives available for colorimetric, fluorimetric and histochemical assays. Several features make the gus gene superior as a reporter gene for plant studies and in the production of genetically engineered crops.
Keywords
Beta D glucuronidase; Beta G1; Beta glucuronidase; FLJ39445; Glucuronidase beta; gurA; GUSB; MPS 7; MPS7; UidA; E. coli Beta Glucuronidase; Escherichia coli Beta Glucuronidase

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References


Protocol for the recovery and detection of Escherichia coli in environmental water samples

ANALYTICA CHIMICA ACTA

Authors: Briciu-Burghina, Ciprian; Heery, Brendan; Regan, Fiona

To achieve active management of bathing areas and to reduce risk associated with the presence of fecal pollution, tests capable of rapid on-site assessment of microbiological water quality are required. A protocol for the recovery and detection of fecal pollution indicator bacteria, E. coli, using beta-glucuronidase (GUS) activity was developed. The developed protocol involves two main steps: sample preparation and GUS activity measurement. In the sample preparation step, syringe filters were used with a dual purpose, for the recovery and pre-concentration of E. coli from the water matrix and as mu L reactors for bacteria lysis and GUS extraction. Subsequently, GUS activity was measured using a continuous fluorometric method developed previously. The optimum GUS recovery conditions for the sample preparation step were found to be 100 mL PELB (supplemented with 1 mg mL(-1) lysozyme and 20 mM DTT) at 37 degrees C for 30 min. The protocol was evaluated on environmental samples (fresh and seawater) against an establish GUS assay method (Coliplage((R))). GUS activities corresponding to samples containing as low as 26 MPN E. coli 100 mL(-1) were detected for the seawater sample and as low as 110 MPN E. coli 100 mL(-1) for the freshwater samples. By comparison with the Coliplage r method, this protocol offered an improvement in the measured GUS activities of 3.1 fold for freshwater samples and 4.1 fold for seawater samples. Furthermore, the protocol developed here, has a time-to-result of 75 min, and successfully addresses the requirement for tests capable of rapid assessment of microbiological water quality. (C) 2017 Elsevier B. V. All rights reserved.

Biosafety of E-coli beta-glucuronidase (GUS) in plants

TRANSGENIC RESEARCH

Authors: Gilissen, LJW; Metz, PLJ; Stiekema, WJ; Nap, JP

The beta-glucuronidase (GUS) gene is to date the most frequently used reporter gene in plants. Marketing of crops containing this gene requires prior evaluation of their biosafety. To aid such evaluations of the GUS gene, irrespective of the plant into which the gene has been introduced, the ecological and toxicological aspects of the gene and gene product have been examined. GUS activity is found in many bacterial species, is common in all tissues of vertebrates and is also present in organisms of various invertebrate taxa. The transgenic GUS originates from the enterobacterial species Escherichia coli that is widespread in the vertebrate intestine, and in soil and water ecosystems. Any GUS activity added to the ecosystem through genetically modified plants will be of no or minor influence. Selective advantages to genetically modified plants that posses and express the E. coli GUS transgene are unlikely. No increase of weediness of E. coli GUS expressing crop plants, or wild relatives that might have received the transgene through outcrossing, is expected. Since E. coli GUS naturally occurs ubiquitously in the digestive tract of consumers, its presence in food and feed from genetically modified plants is unlikely to cause any harm. E. coli GUS in genetically modified plants and their products can be regarded as safe for the environment and consumers.

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