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.
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Comprehensive and state-of-the-art, Bioluminescence Methods and Protocols brings together a collection of innovative methods for the use of luciferase and green fluorescent protein in analytical biochemistry, molecular toxicology, environmental monitoring, and cell-based assays. The contributors-all hands-on researchers who have perfected their protcols-illustrate these powerful methods with a wide variety of applications ranging from the analytical determination (titer) of specific molecules to the cellular detection of a wide range of pollutants, to microbiological detection and analysis, and the molecular biological manipulation of lux, luc, and gfp structural genes. Step-by-step methods for the construction of other recombinant organisms are also included.
Bioluminescence Methods and Protocols represents the state-of-the-art in luminescent assays. Its cutting-edge techniques, diversely illustrated applications, and detailed instructions will make it the standard reference for today's investigators who want to adapt established techniques to their purposes or apply them to a wide range of clinical, microbiological, environmental, toxicological, and analytical problems.
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