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

Anuradha Raghunathan; Bernhard O Palsson

doi:10.1023/a:1022497310798

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

Biotechnol Lett. 2003 Mar;25(5):435-41. doi: 10.1023/a:1022497310798.

Authors

Anuradha Raghunathan¹, Bernhard O Palsson

Affiliation

¹ Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093-0412, USA.

PMID: 12882568
DOI: 10.1023/a:1022497310798

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.

Publication types

Evaluation Study
Validation Study

MeSH terms

Adaptation, Physiological / genetics
Biological Evolution
Chromatography, High Pressure Liquid / methods*
DNA Mutational Analysis / methods*
Escherichia coli / genetics
Escherichia coli / physiology*
Evolution, Molecular*
Gene Expression Profiling / methods
Gene Expression Regulation, Bacterial / physiology
Genome, Bacterial
Pilot Projects
Sequence Alignment / methods
Sequence Analysis, DNA / methods