Transformação de amostras de Escherichia coli com plasmídeo contendo a proteína GFP

Abstract

The gut microbiota has a great diversity of genera and species. The most recent studies show an effective participation of the microbiome not only in the modulation and production of virulence factors of bacteria present in the intestine, but also in the signaling and communication between the body's organs. The study models used so far have limitations and do not allow an effective study of identification and quantification of distinct populations of related species. The cellular assays used for the study of biofilm formation and bacterial adhesion are very effective for visualization of samples, but in the case of mixed cultures of species with similar characteristics it is not possible to distinguish. In this study, bacterial transformation was performed by inserting the plasmid containing the green fluorescent protein (GFP) gene, giving the samples the ability to emit green light. After confirmation of the transformation, the samples were used in biofilm formation and bacterial adhesion assays in in vitro epithelial tissues containing more than one bacterial population. It was possible to visualize and identify the different samples with the use of staining and fluorescence microscopy.