Trabajo interesante del grupo de Eduardo Santero.
Centro Andaluz de Biología del Desarrollo, Universidad Pablo de
Olavide/Consejo Superior de Investigaciones Científicas/Junta de
Andalucía, and Departamento de Biología Molecular e Ingeniería
Bioquímica, Universidad Pablo de Olavide
Uso de fósmidos, librerías metagenómicas, detección de especies resistentes a antibióticos.
The extraordinary potential of metagenomic functional analyses to
identify activities of interest present in uncultured microorganisms has
been limited by reduced gene expression in surrogate hosts. We have
developed vectors and specialized E. coli strains as improved
metagenomic DNA heterologous expression systems, taking advantage of
viral components that prevent transcription termination at metagenomic
terminators. One of the systems uses the phage T7 RNA-polymerase to
drive metagenomic gene expression, while the other approach uses the
lambda phage transcription anti-termination protein N to limit
transcription termination. A metagenomic library was constructed and
functionally screened to identify genes conferring carbenicillin
resistance to E. coli. The use of these enhanced expression
systems resulted in a 6-fold increase in the frequency of carbenicillin
resistant clones. Subcloning and sequence analysis showed that, besides
β-lactamases, efflux pumps are not only able contribute to carbenicillin
resistance but may in fact be sufficient by themselves to convey
carbenicillin resistance.
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