Specific GFP-binding artificial proteins (αRep): a new tool for in?vitro to live cell applications

A family of artificial proteins, named αRep, based on a natural family of helical repeat was previously designed. αRep members are efficiently expressed, folded and extremely stable proteins. A large αRep library was constructed creating proteins with a randomized interaction surface. In the present study, we show that the αRep library is an efficient source of tailor-made specific proteins with direct applications in biochemistry and cell biology. From this library, we selected by phage display αRep binders with nanomolar dissociation constants against the GFP. The structures of two independent αRep binders in complex with the GFP target were solved by X-ray crystallography revealing two totally different binding modes. The affinity of the selected αReps for GFP proved sufficient for practically useful applications such as pull-down experiments. αReps are disulfide free proteins and are efficiently and functionally expressed in eukaryotic cells: GFP-specific αReps are clearly sequestrated by their cognate target protein addressed to various cell compartments. These results suggest that αRep proteins with tailor-made specificity can be selected and used in living cells to track, modulate or interfere with intracellular processes.     

Temperature and strain effects on reproduction and survival of Trichogramma oleae and Trichogramma cacoeciae (Hymenoptera: Trichogrammatidae)

To assess differences in temperature sensitivity during development, life tables for two lines derived from the species Trichogramma oleae Voegelé and Pointel and a strain of Trichogramma cacoeciae Marchal (Hymenoptera: Trichogrammatidae) were elaborated at 15, 20, 25, 30, 35, 36, and 37°C in the laboratory. Eggs of Ephestia kuehniella Zeller together with a fresh drop of honey were supplied every 2 days until the death of the test females, and the removed host egg batches were placed in the equivalent rearing cabinet. The line ‘2F’ of T. oleae was found to be the most efficient at any range of temperatures except at 20 and 37°C, in comparison to the other tested strains. For all species, no progeny emerged from eggs incubated at 36°C and none of the parasitized eggs turned black at 37°C. The better performance at a broader range of temperatures by T. oleae (line 2 F) might be caused by a shorter history in artificial rearing in comparison to the other strains. Fewer generations at laboratory conditions and frequent multiplication on eggs of its natural host (the olive moth Prays oleae) may have prevented a deterioration in the rearing population of this strain, maintaining its genetic diversity at a higher scale. Applying varying temperature regimes on the rearing stock at regular intervals during the mass production process may help to maintain the essential quality of the biological control agents for field performance at higher temperatures.     

Study of 'Redhaven' peach and its white-fleshed mutant suggests a key role of CCD4 carotenoid dioxygenase in carotenoid and norisoprenoid volatile metabolism

Background  

Carotenoids are plant metabolites which are not only essential in photosynthesis but also important quality factors in determining the pigmentation and aroma of flowers and fruits. To investigate the regulation of carotenoid metabolism, as related to norisoprenoids and other volatile compounds in peach (Prunus persica L. Batsch.), and the role of carotenoid dioxygenases in determining differences in flesh color phenotype and volatile composition, the expression patterns of relevant carotenoid genes and metabolites were studied during fruit development along with volatile compound content. Two contrasted cultivars, the yellow-fleshed 'Redhaven' (RH) and its white-fleshed mutant 'Redhaven Bianca' (RHB) were examined.