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Biomedical subjects

O Cabré

Publications and source records attributed to O Cabré.

7 recordsLinked to original sources

Molecular study of the germinal reversions induced at the white-ivory locus in Drosophila melanogaster.

The white-ivory somatic mutation test of Drosophila melanogaster is based on the reversion of the X-linked eye colour recessive mutation white-ivory to wild-type. Although the exact mechanism of white-ivory reversion is not quite understood, it has been suggested that such reversion, both in somatic and germ-line cells, could be due to the precise excision of the tandemly duplicated 2.96 kb DNA fragment characteristic of the white-ivory mutation. We have attempted to confirm this hypothesis analysing, at the molecular level, different germinal revertants induced by chemical treatment with three well known alkylating agents: ethyl methanesulphonate, methyl methanesulphonate and N-nitroso-N-ethylurea. The molecular analysis of these germ-line revertants, using Southern blot hybridization and polymerase chain reaction techniques, shows that such reversions are associated with the deletion of the 2.96 kb tandemly duplicated DNA sequence of the white-ivory locus.

Alkylating Agents

Differential aldehyde sensitivity of newly replicated chromatin from Drosophila melanogaster embryos.

By using a series of formalin concentrations we have found that high aldehyde levels in the fixation buffer of Miller spreads are correlated with the appearance of nonnucleosomal stretches in newly replicated chromatin of embryos from Drosophila melanogaster. These nucleosome-free gaps are found 0-500 nm behind the replication fork and do not correspond to naked DNA. The analysis of the distribution of nucleosome-free gaps on newly replicated DNA has revealed some structural details about the maturation of nucleosomes and provides direct evidence that parental nucleosomes have an altered structure at the replication fork. Finally, these stretches of nonnucleosomal chromatin are located in a trans disposition inside the active replicon, although there exists a considerable variability.

Animals

Restriction mapping of phage lambda vectors using non-radioactive methods.

In order to take advantage of non-radioactive methods, we have developed two plasmids (p lambda LE and p lambda RE) for mapping restriction sites of long inserts cloned in phage lambda vectors. These plasmids are constructed by cloning the left 402-bp and right 560-bp phage lambda genome ends, respectively. To map restriction sites, the cloned sequences in p lambda LE and p lambda RE are labeled with digoxygenin and hybridized to partially digested lambda DNA. The ladder of bands detected with these probes can be used to construct restriction maps in the same way as those obtained using radioactively labeled cos complementary oligodeoxyribonucleotides [Rackwitz et al., Gene 30 (1984) 195-200].

Bacteriophage lambda

Effect of cycloheximide on different stages of Drosophila melanogaster.

Cycloheximide, an antibiotic inhibiting protein synthesis, exerted a toxic effect on different developmental stages egg, larva and adult of Drosophila melanogaster. At the egg stage the early embryos were most sensitive. With larvae, a strong decrease in viability was found, with no sex difference. In adults, there was a dose-effect relationship, mortality increasing with concentration. At 10 and 15 mM, males were more sensitive than females. There were consistent differences between the control and cycloheximide-fed females in respect of the average number of eggs deposited and offspring produced.

Animals

Studies on the role and mode of operation of the very-lysine-rich histones in eukaryote chromatin. The conformation of phi1 histones from marine invertebrate sperm.

Proton magnetic resonance, circular dichroism and infrared spectroscopy are used to investigate the secondary and tertiary structure of three very lysine-rich histones from marine invertebrate sperm. At high ionic strength both Arbacia lixula and Holothuria tubulosa histone phi 1 are observed to contain 25-30% alpha-helix, no beta-structure and to form specific folded structures. Both phi 1 proton magnetic resonance spectra have perturbed methyl resonances at chemical shifts close to those observed for calf thymus H1, suggesting analogies in tertiary structure. Mytilus edulis histone phi 1 however, shows no spectroscopic evidence of secondary and tertiary structure on salt addition.

Amino Acids