A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level.
We present a novel method for visualizing intracellular metabolite concentrations within single cells of Escherichia coli and Corynebacterium glutamicum that expedites the screening process of producers. It is based on transcription factors and we used it to isolate new L-lysine producing mutants of C. glutamicum from a large library of mutagenized cells using fluorescence-activated cell sorting (FACS). This high-throughput method fills the gap between existing high-throughput methods for mutant generation and genome analysis. The technology has diverse applications in the analysis of producer populations and screening of mutant libraries that carry mutations in plasmids or genomes.
Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany.
Stephan
S
Binder
Georg
G
Schendzielorz
Norma
N
Stäbler
Karin
K
Krumbach
Kristina
K
Hoffmann
Michael
M
Bott
Lothar
L
Eggeling
D048230:Corynebacterium glutamicum / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005434:Flow Cytometry; D015723:Gene Library; D005798:Genes, Bacterial; D014644:Genetic Variation; D016680:Genome, Bacterial; D023281:Genomics / Q000379:methods*; D008239:Lysine / Q000235:genetics / Q000378:metabolism
D008239:Lysine