Biotechnol Lett. 2003 Mar;25(5):435-41.

Scalable method to determine mutations that occur during adaptive evolution of Escherichia coli.

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Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093-0412, USA.

Abstract

Denaturing HPLC was used to determine mutations occurring during the adaptive evolution of Escherichia coli K-12. The strains were evolved over 700 generations on glycerol as the sole carbon source from a sub-optimal to an optimal growth rate. The mutations detected by direct sequencing of amplicons of the glycerol-phosphate regulon repressor (glpR) gene were a synonymous substitution Val20Val in two separately evolved strains. Non-synonymous substitutions, Val119Gly and Gly179Trp, were also observed in each of the two strains. This procedure can be scaled to determine genome-scale sequence variations that have occurred during adaptive evolution.

PMID:: 12882568; [PubMed - indexed for MEDLINE]

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