Determination of ATP with a Firefly Luciferase-Based System for the Detection of Life in Mars
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Expression and Purification of the luciferase enzyme and in Vivo ATP Assay
Introduction: Gene expression and purification of luciferases from the firefly, Lampyris turkestanicus, and optimization of cellular ATP measurements were performed. Methods: cDNA encoding luciferases from Lampyris turkestanicus was transferred from pQE30 vector into pET28a expression vector and pLtu28 was built. Newly constructed vector was expressed in E. coli XL1 Blue and the recombinant l...
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We expressed and purified a recombinant P. pyralis luciferase with N-terminal His-tags. The silanized Ni or Cu-loaded magnetic particles were prepared and used to assemble the His-tagged P. pyralis luciferase. This enzyme immobilized on functionalized magnetic nanoparticles (MNPs) via electrostatic interactions of His-tag with Ni2+/Cu2+ ions on the surface of MNPs using si...
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Effects of ATP and Mg2+ concentrations on bioluminescence reaction of luciferase (Photinus pyralis) were investigated by home-made luminometer. The Michaelis constants of the enzyme for ATP and Mg2+ obtained from the Lineweaver-Burk graph, were 61.9 mM ±3.3 mM and 251.6mM ± 39.0mM...
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متن کاملDesign and application of a bioluminescent biosensor for detection of toxicity using Huh7-CMV-luc cell line
Cell-based biosensors (CBBs) are becoming important tools for biosecurity applications and rapid diagnostics in food microbiology for their unique capability of detecting hazardous materials. Pollutants, such as heavy metals and chemicals, are now considered as a global threat and are associated with detrimental health outcomes. Fast and accurate detection of pollutants is essential to reduce t...
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