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71.
Branchini BR Ablamsky DM Murtiashaw MH Uzasci L Fraga H Southworth TL 《Analytical biochemistry》2007,361(2):253-262
Light emission from the North American firefly Photinus pyralis, which emits yellow-green (557-nm) light, is widely believed to be the most efficient bioluminescence system known, making this luciferase an excellent tool for monitoring gene expression. We present studies on the production of a set of thermostable red- and green-emitting luciferase mutants with bioluminescent properties suitable for dual-color reporter assays, biosensor measurements with internal controls, and imaging techniques. Starting with the luciferase variant Ser284Thr, we introduced the mutations Thr214Ala, Ala215Leu, Ile232Ala, Phe295Leu, and Glu354Lys to produce a new red-emitting enzyme with a bioluminescence maximum of 610 nm, narrow emission bandwidth, favorable kinetic properties, and excellent thermostability at 37 degrees C. By adding the same five changes to luciferase mutant Val241Ile/Gly246Ala/Phe250Ser, we produced a protein with an emission maximum of 546 nm, providing a set of thermostable enzymes whose bioluminescence maxima were separated by 64 nm. Model studies established that the luciferases could be detected at the attomole level and six orders of magnitude higher. In microplate luminometer format, mixtures containing 1.0 fmol total luciferase were quantified from measurements of simultaneously emitted red and green light. The results presented here provide evidence that it is feasible to monitor two distinct activities at 37 degrees C with these novel thermostable proteins. 相似文献
72.
A number of xenobiotics are toxic because they rcdox cycle and generate free radicals. Interaction with iron, either to produce reactive species such as the hydroxyl radical, or to promote lipid peroxidation, is an important factor in this toxicity. A potential biological source of iron is ferritin. The cytotoxic pyrimidines, dialuric acid, divicine and isouramil, readily release iron from ferritin and promote ferritin-dependent lipid peroxidation. Superoxide dismutase and GSH, which maintain the pyrimidines in their reduced form, enhance both iron release and lipid peroxidation. Microsomes plus NADPH can reduce a number of iron complexes, although not ferritin. Reduction of Adriamycin. paraquat or various quinones to their radicals by the microsomes enhances reduction of the iron complexes, and in some cases, enables iron release from ferritin. Adriamycin stimulates iron-dependent lipid peroxidation of the microsomes. Ferritin can provide the iron, and peroxidation is most pronounced at low PO2. Compiexing agents that supress intraccllular iron reduction and lipid peroxidation may protect against the toxicity of Adriamycin. 相似文献
73.
Hugo Jose Garavelli 《Mycopathologia》1964,22(1):40-46
Resumen Se describe el aislamiento delKeratinomyces ajelloi en 5 sobre un total de 105 muestras de tierras de la ciudad de Buenos Aires y alrededores. Se hace ademas una reseña sobre el método empleado para el aislamiento de dicho hongo (método deVanbreuseghem) y un resumen sobre los conocimientos actuales de esta especie descubierta porVanbreuseghem en 1952 utilizando su método. 相似文献
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Anake Kijjoa Astréa M. Giesbrecht Otto R. Gottlieb Hugo E. Gottlieb 《Phytochemistry》1981,20(6):1385-1388
The trunk woods of two Amazonian Myristicaceae, Virola minutiflora and V. elongata, contain respectively 1-(2-hydroxy-4-methoxyphenyl)-3-(3,4-m 相似文献
79.
Marden A. De Alvarenga J. Jerônimo Da Silva Hugo E. Gottlieb Otto R. Gottlieb 《Phytochemistry》1981,20(5):1159-1161
The structure of micrandrol-C from Micrandropsis scleroxylon (Euphorbiaceae) is revised to 2,6-dihydroxy-7-methyl-1-methylthiophenanthrene. This and other micrandrols are probably diterpenes in view of their co-occurrence with micrandrol-D, the hemiketal of 1,2,3,4,9,10-hexahydro-6-hydroxy-4a-hydroxymethyl-1,1,7-trimethy-2-oxophenanthrene. 相似文献
80.
Because of their rarity, primary malignant tumors of the heart continue to evade clinical preoperative diagnosis. In five cases of malignant heart tumor that were reviewed from the diagnostic point of view, the clinical symptoms and radiographic abnormalities were found to be similar to those produced by more common heart diseases. The diagnosis of malignant cardiac tumor should be considered in a patient with heart disease who has (1) an unusual cardiomediastinal silhouette, (2) an atypical cardiac calcification, and (3) clinical or radiographic cardipulmonary findings which do not conform to the expected behavior of the common forms of heart disease. 相似文献