RESUMEN
Cell competition, the conditional loss of viable genotypes only when surrounded by other cells, is a phenomenon observed in certain genetic mosaic conditions. We conducted a chemical mutagenesis and screen to recover new mutations that affect cell competition between wild-type and RpS3 heterozygous cells. Mutations were identified by whole-genome sequencing, making use of software tools that greatly facilitate the distinction between newly induced mutations and other sources of apparent sequence polymorphism, thereby reducing false-positive and false-negative identification rates. In addition, we utilized iPLEX MassARRAY for genotyping recombinant chromosomes. These approaches permitted the mapping of a new mutation affecting cell competition when only a single allele existed, with a phenotype assessed only in genetic mosaics, without the benefit of complementation with existing mutations, deletions, or duplications. These techniques expand the utility of chemical mutagenesis and whole-genome sequencing for mutant identification. We discuss mutations in the Atm and Xrp1 genes identified in this screen.
Asunto(s)
Mapeo Cromosómico , Drosophila melanogaster/efectos de los fármacos , Drosophila melanogaster/genética , Metanosulfonato de Etilo/farmacología , Genoma de los Insectos , Genotipo , Secuenciación de Nucleótidos de Alto Rendimiento , Mutación/efectos de los fármacos , Alelos , Animales , Proteínas de la Ataxia Telangiectasia Mutada/genética , Reparación del ADN , Estudios de Asociación Genética , Pruebas Genéticas , Mutagénesis , Fenotipo , Carácter Cuantitativo Heredable , Recombinación GenéticaRESUMEN
Recently, different bacteriological laboratory interventions that decrease reporting time have been developed. These promising new broad-based techniques have merit, based on their ability to identify rapidly many bacteria, organisms difficult to grow or newly emerging strains, as well as their capacity to track disease transmission. Maldi-TOF MS has been proven to be an accurate and reliable method for organism identification including bacteria, yeast, molds, and mycobacteria. It is rapid, with results often 24 hours earlier than traditional methods, and inexpensive. The range of applications of Maldi-TOF MS has been growing constantly, from rapid species identification to labor-intensive proteomic studies of bacterial physiology (bacterial resistance and virulence). The purpose of this review is to present the different solutions commercialized in France, summarize the place of this technology in microbiology lab and to analyze future perspectives in this field.