Isolation and characterization of virulence and resistance genes from the dental biofilm metagenome by heterologous expression in Escherichia coli

Abstract

The highly complex microbiota harbored by dental biofilm, along with the various clinical patterns of periodontitis, highlights the necessity of more comprehensive investigations on the pathogenicity mechanisms engendered by this oral pathology. Moreover, the emergence of novel antibiotic resistant bacterial lineages points to the importance of studies targeting the development of new strategies for controlling bacterial dissemination. Previous studies have shown that metagenomic analysis based on heterologous expression techniques can unveil novel biomolecules with medical, dental and biotechnological importance. The aim of the present study is the isolation and molecular characterization of virulence and resistance genes from the periodontopathic dental biofilm metagenome. In this study, a total of 20 periodontitis patients will be analised. The metagenomic DNA will be extracted, cloned into plasmid vector, transformed in Escherichia coli, heterologous proteins will be expressed. Bacterial colonies with differential growth activities will have their plasmid DNA extracted and sequenced. DNA sequences will be submitted to computational analyzes with public domain databases. In all, the study aims to isolate, identify and characterize potentially new biomolecules which may contribute to a better understanding of the pathogenic mechanisms in periodontitis and, hence, to the development of specif therapeutic strategies for this oral pathology. (AU)