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I Djajanegara R Holtzapffel P M Finnegan M H Hoefnagel D A Berthold J T Wiskich D A Day 《FEBS letters》1999,454(3):220-224
The alternative oxidase is a quinol oxidase of the respiratory chain of plants and some fungi and protists. Its activity is regulated by redox-sensitive disulphide bond formation between neighbouring subunits and direct interaction with certain alpha-ketoacids. To investigate these regulatory mechanisms, we undertook site-directed mutagenesis of soybean and Arabidopsis alternative oxidase cDNAs, and expressed them in tobacco plants and Escherichia coli, respectively. The homologous C99 and C127 residues of GmAOX3 and AtAOX1a, respectively, were changed to serine. In the plant system, this substitution prevented oxidative inactivation of alternative oxidase and rendered the protein insensitive to pyruvate activation, in agreement with the recent results from other laboratories [Rhoads et al. (1998) J. Biol. Chem. 273, 30750-30756; Vanlerberghe et al. (1998) Plant Cell 10, 1551-1560]. However, the mutated protein is instead activated specifically by succinate. Measurements of AtAOX1a activity in bacterial membranes lacking succinate dehydrogenase confirmed that the stimulation of the mutant protein's activity by succinate did not involve its metabolism. Examples of alternative oxidase proteins with the C to S substitution occur in nature and these oxidases are expected to be activated under most conditions in vivo, with implications for the efficiency of respiration in the tissues which express them. 相似文献
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Summary Spontaneous cell-to-cell transformation between naturally competent bacteria on selective media resulted in an overestimation of the transferability of genetic information. EDTA effectively prevented transformation on selective media whereas DNaseI did not reliably inhibit cell-to-cell transformation. An improved method to estimate gene transfer frequencies is described. 相似文献
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Valuable biological information can be obtained by monitoring the movement of organisms. However, the choice of monitoring
method becomes highly restricted when following small organisms (<100 mm), especially in aquatic ecosystems. Stable isotopes
are being increasingly used in this respect but rarely at the local spatial scale, i.e. 10–1000 s of metres. We sought to
identify movement of small fishes between a main river channel and its tributary. Little overlap in isotope baseline was detected
between the two channels despite some temporal variability in δ15N of baseline indicator organisms in the main river. The individuals of two small cyprinid fish species (Leuciscus souffia and Alburnoides bipunctatus) of all the size classes (40–100 mm) caught within the tributary showed considerable heterogeneity in δ15N values. Classification and discriminant analysis on isotope-derived data distinguished two significantly different groups.
Moreover, this result was supported by further sampling of fish caught in the main river (in May and December 2006). Alternative
hypotheses, such as dietary differences, biological factors, temporal shifts and spatial differences in diet, did not explain
δ15N variability. This application of stable isotopes at a relatively small spatial and temporal scales further demonstrates
its potential as a tool for ecologists. 相似文献