Novel molecular tools to study mycoplasma cells

• Autores: Ana María Martínez Mariscal
• Directores de la Tesis: Enrique Querol Murillo (dir. tes.), Jaume Piñol Ribas (dir. tes.)
• Lectura: En la Universitat Autònoma de Barcelona ( España ) en 2017
• Idioma: español
• Tribunal Calificador de la Tesis: Francesc Xavier Avilés Puigvert (presid.), Maria Lluch Senar (secret.), Alicia Broto Hernández (voc.)
• Programa de doctorado: Programa de Doctorado en Bioquímica, Biología Molecular y Biomedicina por la Universidad Autónoma de Barcelona
• Materias:
  • Química
    • Bioquímica
  • Ciencias de la vida
    • Genética
      • Ingeniería genética
• Enlaces
  • Tesis en acceso abierto en: TESEO
• Resumen

  • Mycoplasmas are human pathogens characterized by their reduced size and for being the free-living organisms with the smallest genomes known. Despite their apparent genomic simplicity, they present a much higher complexity that requires the most advanced molecular techniques to be studied and understood in their entirety. But the tools currently available for the modification of mycoplasma cells are reduced to a small repertoire based on minitransposons, replicative plasmids for some species and intrinsic homologous recombination in some species and specific strains. For that reason, this thesis has developed and implemented techniques that allow the modification of these cells, making available new tools that allow analysis that were limited to other bacteria until now.

    New knowledge on the locations of different proteins belonging to the Mycoplasma genitalium terminal organelle, a key structure in the motility and pathogenicity of this microorganism, is provided in this work. In addition, this work reinforces the usefulness of fluorescence proteins as molecular markers for the study of M. genitalium.

    On the other hand, the Cre-lox system has been implemented in M. genitalium , allowing deletions without leaving more marks than 34 bp of the lox sequences. Moreover, this technology also allows the elimination of antibiotics resistance gene markers.

    Finally, iCRISPR interference system has been implemented in Mycoplasma pneumoniae, allowing for the first time the downregulation of genes in a specific way. In addition, the implementation of this technique opens the doors to the study of essential genes that until now was not possible in this microorganism.