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

Authors: Raghunathan A.¹; Palsson B.Ø.^{1, 2, 3}

Source: Biotechnology Letters, Volume 25, Number 5, March 2003 , pp. 435-441(7)

Publisher: Springer

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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.

Keywords: adaptive evolution; denaturing HPLC; Escherichia coli; mutation sequence variation; phenotype

Language: English

Document Type: Research article

Affiliations: 1: Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093-0412, USA 2: (Fax: +1-858-822-3120 3: E-mail: palsson@ucsd.edu)

Publication date: 2003-03-01