tailieunhanh - Bioluminescence Recent Advances in Oceanic Measurements and Laboratory Applications Part 11

Tham khảo tài liệu 'bioluminescence recent advances in oceanic measurements and laboratory applications part 11', khoa học tự nhiên, công nghệ sinh học phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | Bioluminescence Applications in Preclinical Oncology Research 141 luciferins that are currently used in biological research. Luciferase is the general term for a class of enzymes which catalzye oxidation reactions involving luciferins. The structure of luciferase seems to have an impact on the wavelength of photons emitted and color of light produced so that reactions may exhibit yellow-green to red light. Although many of the luciferase enzymes discussed in Table 1 have been applied to in vitro and in vivo work firefly luciferase F-Luc is most commonly employed in oncology research. In the chemical reaction between F-Luc and its substrate D-luciferin ATP and oxygen promote the formation of oxy-luciferin species. The emission spectrum of the F-Luc catalyzed reaction is broad with an emission peak at approximately 560 nm and a large component above 600 nm. Yellow-green to yellow-orange light is emitted following the relaxation of oxy-luciferin to its ground state see reaction 2 . Reaction 2 D-Luciferin ATP reversible uses luciferase and Mg2 luciferase luciferyl-AMP PPi Luciferyl-AMP oxygen uses luciferase and Mg2 luciferase oxyluciferin AMP CO2 hv de Wet Wood et al. 1987 3. Constructing bioluminescent mammalian cells and animal models In the following sections the application of biological models expressing luciferase are described in detail. Mammalian systems do not naturally express luciferase and therefore these systems must be engineered to expresse the enzyme. A vector capable of constitutive expression of luciferase or vectors designed to achieve controlled expression of the enzyme can be used Fusion genes can be used as reporter constructs to track gene delivery and integration. Expression of fusion proteins can be used to follow protein localization and protein-protein interactions. Cells with stable expression of luciferase can be used for in vitro assays or be injected into animals to examine metabolism immunity angiogenesis or to establish disease that .