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Low-mutation-rate, reduced-genome Escherichia coli: an improved host for faithful maintenance of engineered genetic constructs

Csörgo, Bálint ; Fehér, Tamás ; Tímár, Edit ; Blattner, Frederick R ; Pósfai, György

Microbial cell factories, 2012-01, Vol.11 (1), p.11-11, Article 11 [Periódico revisado por pares]

England: BioMed Central Ltd

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Título:
Low-mutation-rate, reduced-genome Escherichia coli: an improved host for faithful maintenance of engineered genetic constructs
Autor: Csörgo, Bálint ; Fehér, Tamás ; Tímár, Edit ; Blattner, Frederick R ; Pósfai, György
Assuntos: Bacteria ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Cell culture ; chassis ; DNA polymerases ; DNA-directed DNA polymerase ; DNA-Directed DNA Polymerase - metabolism ; DNA-Directed RNA Polymerases - metabolism ; Enzymes ; Escherichia coli ; Escherichia coli - enzymology ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Evolution ; evolvability ; Genetic aspects ; Genetic diversity ; Genetic Engineering ; Genetic variation ; Genome, Bacterial ; Genomes ; Industrial applications ; Methyltransferase ; Microbiology ; Molecular modelling ; Mutagenesis ; Mutation ; Mutation Rate ; Mutation rates ; Physiological aspects ; Plasmids ; reduced genome ; SOS Response (Genetics) ; Stress ; synthetic biology ; Viral Proteins - metabolism
É parte de: Microbial cell factories, 2012-01, Vol.11 (1), p.11-11, Article 11
Notas: ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
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Descrição: Molecular mechanisms generating genetic variation provide the basis for evolution and long-term survival of a population in a changing environment. In stable, laboratory conditions, the variation-generating mechanisms are dispensable, as there is limited need for the cell to adapt to adverse conditions. In fact, newly emerging, evolved features might be undesirable when working on highly refined, precise molecular and synthetic biological tasks. By constructing low-mutation-rate variants, we reduced the evolutionary capacity of MDS42, a reduced-genome E. coli strain engineered to lack most genes irrelevant for laboratory/industrial applications. Elimination of diversity-generating, error-prone DNA polymerase enzymes involved in induced mutagenesis achieved a significant stabilization of the genome. The resulting strain, while retaining normal growth, showed a significant decrease in overall mutation rates, most notably under various stress conditions. Moreover, the error-prone polymerase-free host allowed relatively stable maintenance of a toxic methyltransferase-expressing clone. In contrast, the parental strain produced mutant clones, unable to produce functional methyltransferase, which quickly overgrew the culture to a high ratio (50% of clones in a 24-h induction period lacked functional methyltransferase activity). The surprisingly large stability-difference observed between the strains was due to the combined effects of high stress-induced mutagenesis in the parental strain, growth inhibition by expression of the toxic protein, and selection/outgrowth of mutants no longer producing an active, toxic enzyme. By eliminating stress-inducible error-prone DNA-polymerases, the genome of the mobile genetic element-free E. coli strain MDS42 was further stabilized. The resulting strain represents an improved host in various synthetic and molecular biological applications, allowing more stable production of growth-inhibiting biomolecules.
Editor: England: BioMed Central Ltd
Idioma: Inglês

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Low-mutation-rate, reduced-genome Escherichia coli: an improved host for faithful maintenance of engineered genetic constructs

Csörgo, Bálint ; Fehér, Tamás ; Tímár, Edit ; Blattner, Frederick R ; Pósfai, György

England: BioMed Central Ltd

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