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Etude de l'organisation transcriptionnelle et de la régulation post-transcriptionnelle du groupe d'ORF codant l'érythrose 4-phosphate déshydrogénase, la phosphoglycérate kinase et la Fructose-1,6-bisphosphate aldolase de classe II chez Escherichia coli : Applications à la production de protéines recombinantes

Abstract : The Erythrose-4-phosphate déshydrogénase (E4PDH)-phosphoglycerate kinase (PGK)fructose-1,6-bisphosphate aldolase (FBA) ORF cluster, corresponding to three enzymes from carbon metabolism, is found in a part of the g-proteobacteria. We showed that, in Escherichia coli, this ORF cluster is an operon with one strong regulated promoter epd P0 and one extended promoter pgk P1. The different transcripts are maturated by RNase E, in epd-pgk and pgk-fbaA region. This result in the differential stability of the transcripts, with by increasing half-life, epd,pgk and fbaA. By continuous fermentation, we showed that epd, pgk, fbaA and gapA expression increases with the growth rate. The low production of acetic acid form by strains with an inactivated transport protein EIIBCGlc led us to test whether they can be used for recombinant protein production. We developed conditions to get high cellular density and high level of recombinant protein production.
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Corinne Vallet. Etude de l'organisation transcriptionnelle et de la régulation post-transcriptionnelle du groupe d'ORF codant l'érythrose 4-phosphate déshydrogénase, la phosphoglycérate kinase et la Fructose-1,6-bisphosphate aldolase de classe II chez Escherichia coli : Applications à la production de protéines recombinantes. Biologie moléculaire. Université Henri Poincaré - Nancy 1, 2005. Français. ⟨NNT : 2005NAN10191⟩. ⟨tel-01748075⟩

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