RT info:eu-repo/semantics/masterThesis
T1 Optimización de un sistema para cuantificar la eficiencia de la conjugación bacteriana in vitro basado en selección antibiótica
A1 Pujol Olivares, Jordina
A2 Máster Univ. en Investigación y Diagnóstico de Enfermedades Tropicales
AB Antibiotic resistance has become one of the major problems in health care nowadays. Bacteriacan acquire antibiotic resistance by different natural mechanisms, but the most important is theacquisition through horizontal gene transfer (HGT). Despite HGT can occur in three mainmechanisms, conjugation is the one that causes the greatest propagation of antibiotic resistancegenes. Conjugation is based on the transmission of conjugative plasmids which carry thesegenes. One of the most promising strategies to fight antibiotic resistance is the investigation ofconjugation inhibitors (COINs).The aim of this study was the optimization of an in vitro conjugation protocol for thereproducible quantification of conjugation frequencies, which permits the study of anticonjugation properties of candidate chemical compounds. Also, we tried to confirm theconstitutively expression of the green fluorescent protein (GFP) in the transconjugant straingenerated.The optimized protocol using nitrocellulose filters and antibiotic selection, employing the donorE. coli MDS42 strain carrying plasmid pJC01 and the recipient E. coli BL21 (DE3) strain, allowedto obtain reproducible conjugation frequencies in the order of 10-1. Additionally, the system wasuseful for the preliminary study of the possible conjugation inhibitory activity of five candidatechemical compounds. Finally, the pJC01 plasmid in the transconjugant strain was stable underantibiotic selection and constitutively express the GFP. This strain will be used for theoptimization of a high-throughput in vitro conjugation protocol for the rapid and cost-effectiveevaluation of COIN candidates
YR 2022
FD 2022
LK http://riull.ull.es/xmlui/handle/915/31848
UL http://riull.ull.es/xmlui/handle/915/31848
LA es
DS Repositorio institucional de la Universidad de La Laguna
RD 21-may-2024