Ponente
Descripción
In genotoxic studies, only one experimental model or assay is not always enough to sense several levels of biological organization, in which the effects of the genetic damages could be expressed. Sometimes the proper characteristics of the organism employed, implies restrictions to the genotoxic assay for evaluating different endpoints. Results obtained from combined assays after a unique exposure, provides the researcher i) the advantage of performing more widespread genotoxic evaluations with less costs and ii) contributes to facilitate the analysis of the possible mechanistic pathways. The ultraviolet spectrum of sunlight is the first environmental mutagen worldwide. Understanding the biological effects of natural irradiations is mandatory and biological sensors are needed. The cosmopolitan organism Caulobacter crescentus is an stablished model to study cell cycle and mutagenesis. In this work is proposed a modified workflow that allows the simultaneous detection of three genotoxic endpoints: cytotoxicity, primary DNA damage and genic mutation. In this sense Survival assay, SOS Chromotest and RifR mutagenesis were assayed. The results shown that the presence of plasmid p3213 did not affect the survival and mutagenesis parameters previously reported in the wild strain. The use of M2 transparent medium is mandatory to study UV impacts, given the absorptive capacity of the PYE. This protocol saves resources and experimental time as simultaneously inform multiple endpoints, an aspect that is mandatory for sensing sunlight unique exposure effects. Finally, this tool could also be used in the evaluation of other types of agents with a significant reduction in amount of product.
Keywords: Biodosimeter, SOS Chromotest, RifR mutagenesis, Survival assay, genotoxic assay
