Scalable method to determine mutations that occur during adaptive evolution of Escherichia coli.
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.
Raghunathan
Anuradha
A
Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093-0412, USA.
Palsson
Bernhard O
BO
D000222:Adaptation, Physiological / Q000235:genetics; D005075:Biological Evolution; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D019143:Evolution, Molecular; D020869:Gene Expression Profiling / Q000379:methods; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D016680:Genome, Bacterial; D010865:Pilot Projects; D016415:Sequence Alignment / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods