LaRossa Bioluminescence Methods and Protocols

Erscheinungsjahr 2008
ISBN: 978-1-59259-577-8
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 102, 310 Seiten, eBook

Reihe: Methods in Molecular Biology

The biological emission of light has fascinated humankind for mill- nia; who is not enchanted by the flashing of fireflies during summer's e- nings? Their collection by children in the area surrounding Baltimore, Maryland in the middle of this century for the purpose of biochemical isolation and ch- acterization of light producing proteins and cofactors may constitute the i- tiation of serious, quantitative studies of this phenomenon. The biochemistry of light production is certainly well-understood today. In bacterial systems it is coupled to the reducing and energy states of the cell, whereas in eukaryotic systems it is dependent upon the availability of ATP. These differences are reflected in the unrelated substrate requirements and structures of eukaryotic and prokaryotic luciferases. These biochemical studies have been aided by the cloning of luciferase-encoding structural genes, the structural genes for cofactor generation, and genes encoding proteins that interact with these lig- producing enzymes. These genes also provide starting materials for a wide range of applications that are the subject of this volume. Many of these advances have earlier been catalogued in two volumes of the Methods in Enzymology series. It is thus necessary to provide a rationale for the publication of Bioluminescence Methods and Protocols. The format of the Methods in Molecular Biology series differs significantly from that of the above-mentioned, classic compilation.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

LaRossa, Robert A.

Inhaltsverzeichnis

The Basics.- Improvements in the Application of Firefly Luciferase Assays.- Visualization of Bioluminescence.- Microscopic Imagery of Mammalian Cells Expressing an Enhanced Green Fluorescent Protein Gone.- Analytical Biochemistry.- Luminometric Measurement of Malate and Glucose-6-Phosphate in Mammalian Tissue.- Bioluminescent Assay of the Guanylates.- Bioluminescent Assay of the Adenylate Energy Charge.- Molecular Biology Tools.- Photorhabdus luminescens luxCDABE Promoter Probe Vectors.- Insertion of Promoter Region::luxCDABE Fusions into the Escherichia coil Chromosome.- Probing for Promoters with Luciferase-Transposons.- Cryopreservation and Reawakening.- Luciferase Renaturation Assays of Chaperones and Chaperone Antagonists.- Cell-Based Assays.- Genotoxic Sensors.- Stress Detection Using Bioluminescent Reporters of the Heat-Shock Response.- Real-Time Reporter of Protein Synthesis Inhibition.- Luminescence-Based Cell Viability Testing.- Phagocyte Chemiluminescence.- Detection of Oxidants Using lux Fusions to Oxidative Stress Promoters.- Environmental Applications.- Luciferase-Based Measurement of Water Contaminants.- Bioluminescence-Based Metal Detectors.- Luminescence Facilitated Detection of Bioavailable Mercury in Natural Waters.- A Panel of Stress-Responsive Luminous Bacteria for Monitoring Wastewater Toxicity.- Organic Contaminant Detection and Biodegradation Characteristics.- Detection of Firefly Luciferase-Tagged Bacteria in Environmental Samples.- Monitoring of GFP-Tagged Bacterial Cells.