Acquisition of a DNA sequencer for the study of gene polymorphisms in microorganisms, the repertoire of T and B cells, and other genes of interestin heart disease

Abstract

The use of state-of-the-art automated DNA sequencers, which employ capillary electrophoresis, presents a series of advantages in relation to that of microplate readers. The sequencing process is almost entirely automated, without the need to prepare plates or gels, or to adjust the sequenced samples after electrophoretic separation. In addition, the runtime in a capillary-based system is shorter, allowing a greater number of samples to be sequenced within a given time frame. This technique has various applications in the study of DNA (sequencing, the study of polymorphisms, and the analysis of fragments, as well as other applications, depending on the software employed). The multi-user device requested will directly aid in the genotyping of microorganisms and of polymorphisms in the repertoire of T and B cells, as well as of other genes of interest in heart disease. By the end of the project, we expect to have gained a better understanding of the distribution of the most prevalent strains of Streptococcus pyogenes in the state of São Paulo, allowing us to evaluate the efficiency of the vaccine in development, which is aimed at preventing the rheumatic fever and other pathologies resulting from infection with S. pyogenes. The sequencer will also facilitate the identification of single-nucleotide polymorphisms in Chagas disease and of alterations in the repertoire of T and B cells in the peripheral blood of patients who have undergone transplantation (solid organ transplant or cell therapy). In addition, the equipment will be employed for the structural and functional analysis of the LEE region and of fimbrial and afimbrial adhesins of atypical enterogenic Escherichia coli. It will also be a valuable tool in the evaluation of various genes cloned in vectors, the objective being to determine the integrity of the DNA and the position of the insert, as well as to identify the mutations induced in genes of interest, for a variety of functional studies and for the production of recombinant proteins. (AU)