Self-assembling protein microarrays

Protein microarrays provide a powerful tool for the study of protein function. However, they are not widely used, in part because of the challenges in producing proteins to spot on the arrays. We generated protein microarrays by printing complementary DNAs onto glass slides and then translating target proteins with mammalian reticulocyte lysate. Epitope tags fused to the proteins allowed them to be immobilized in situ. This obviated the need to purify proteins, avoided protein stability problems during storage, and captured sufficient protein for functional studies. We used the technology to map pairwise interactions among 29 human DNA replication initiation proteins, recapitulate the regulation of Cdt1 binding to select replication proteins, and map its geminin-binding domain.     

Heritability and predicted gain from selection in components of crop duration in divergent chickpea cross populations

Genetic analysis of ten quantitative traits related to crop duration in chickpea was carried out using three F ₂ sib-populations; 272-2 × CDC Anna, 298T-9 × CDC Anna and 298T-9 × CDC Frontier. F ₃ and F ₄ families from these populations were further evaluated for traits found important in the initial study. Also, 112 recombinant inbred lines (RILs) of chickpea cross ICCV 2 × JG 62 were evaluated for days to flowering, days to maturity and reproductive period. An analysis of the F ₂ population data using the mixed model approach revealed that the additive component of variance was significant for days to flowering, days to first podding and days to first pod maturity, while dominance genetic variance was significant for morphological components of crop duration such as height to first pod and height at flowering. Comparatively high heritability estimates (39–48%) were obtained for days to flowering, days to first pod maturity, percent pod maturity at four months after planting and days to maturity based on offspring-parent (F ₄ and F ₃ generations) regression and/or analysis of variance for the RIL population. The predicted gain from selection as a percentage of the population mean was low (5% or less) for these key components of crop duration owing to the low variability detected within the populations, the exception being percent pod maturity. To maximize gain from selection in these traits, it is therefore, essential to increase genetic variability among the progenies, potentially through multi-parent crosses that may involve gene introgression from across desi and kabuli types of chickpea and from wild progenitors.     

Antagonism of pheromone response of Ostrinia nubilalis males and implications on behavior in the laboratory and in the field

The antagonistic effect on the pheromone response and catabolism of male European corn borers, Ostrinia nubilalis, by several trifluoromethyl ketones is reported. (Z)-11-Tetradecenyl trifluoromethyl ketone (Z11-14:TFMK), the most closely related analogue of the main component of the pheromone, elicits a remarkable disruptive effect on close approach and source contact of males flying to a source baited with mixtures of the pheromone and the antagonist in 5:1 and 10:1 ratios. In this experiment, the male displayed an erratic flight track with frequent counter turns and intersections with the plume. In the field, the TFMK significantly lowered the number of males caught when mixed with the pheromone in a 10:1 ratio in comparison with the natural attractant. The compound was also a good inhibitor of the antennal esterase of the insect with a IC(50) value of 0.28 muM. The homologous (Z)-10-tridecenyl trifluoromethyl ketone, with one carbon less in the chain, also elicited an antagonistic effect in the wind tunnel, but in the field, the results were not conclusive. The effect induced was lower than the one displayed by Z11-14:TFMK including the activity as the esterase inhibitor (IC(50) value of 7.55 muM). The saturated tetradecyl trifluoromethyl ketone, tetradecyltrifluoropyruvamide, and (Z)-11-2-thiatetradecenyl trifluoromethyl ketone resulted completely inactive. The results obtained in conjunction to the previously shown low toxicity to mice by related trifluoromethyl ketones provide new important data for the putative utilization of these chemicals as new pest control agents.     

Production and characterization of monoclonal antibodies against midgut of ixodid tick,Haemaphysalis longicornis

There are concerted efforts toward development of tick vaccines to replace current chemical control strategies that have serious limitations [Parasitologia 32 (1990) 145; Infectious Disease Clinics of North America (1999) 209-226]. In this study, monoclonal antibodies (mAbs) specific to Haemaphysalis longicornis midgut proteins were produced and characterized. Eight antibody-secreting hybridomas were cloned and the mAbs typed as IgG1, IgG2a and IgG2b. On immunoblots, all mAbs reacted with a midgut protein band of about 76 kDa. All mAbs uniformly immunogold-stained the surface or epithelial layers of H. longicornis midgut and endosomes. Adult ticks (50%) that fed on an ascitic mouse producing the IgGs developed a red coloration and did not oviposit. As such, the 76 kDa protein that reacted with the mAbs could, therefore, be a potential candidate for tick vaccine development.     

Field-testing and characterization of Technosols made from industrial and agricultural residues for restoring degraded slopes in semiarid SE Spain

Journal of Soils and Sediments - Many industries and agriculture generate huge amounts of wastes either mineral, organic or both, with environmental impact. Some of these residues have the...     

Effect of castration at 10 months of age on growth physiology and behavior of male feral beef cattle

This study compared the growth performance, plasma testosterone and cortisol levels around castration at 10 months of age, and plasma insulin‐like growth factor (IGF)‐I concentration and flight speed, in intact bulls and steers from 10 to 21 months of age in a feral Spanish breed. Fourteen bulls (366.5 ± 48.5 kg live weight) were assigned at random to one of two treatments: surgically castrated (steers) or intact (bulls), and submitted to an identical fattening period. Steers reared until heavy live weights (21 months of age) grew slowly and had lower plasma IGF‐I concentrations than intact bulls. These differences were mainly highlighted the month after surgery (11 months of age) and the last part of the fattening period (from 19 to 21 months of age). After surgical castration (11 and 12 months of age), steers showed a tendency to display greater flight speed values than intact bulls but baseline plasma cortisol concentration did not differ between groups at this time. At the end, steers and bulls reached nearly similar temperament, as flight speed did not differ between them. The results confirm the role of IGF‐I as a key anabolic hormone in male beef cattle and thus it may reflect growth differences due to altered sex steroids production.