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Genome-scale analysis of anti-metabolite directed strain engineering.

Publication ,  Journal Article
Bonomo, J; Lynch, MD; Warnecke, T; Price, JV; Gill, RT
Published in: Metabolic engineering
March 2008

Classic strain engineering methods have previously been limited by the low-throughput of conventional sequencing technology. Here, we applied a new genomics technology, scalar analysis of library enrichments (SCALEs), to measure >3 million Escherichia coli genomic library clone enrichment patterns resulting from growth selections employing three aspartic-acid anti-metabolites. Our objective was to assess the extent to which access to genome-scale enrichment patterns would provide strain-engineering insights not reasonably accessible through the use of conventional sequencing. We determined that the SCALEs method identified a surprisingly large range of anti-metabolite tolerance regions (423, 865, or 909 regions for each of the three anti-metabolites) when compared to the number of regions (1-3 regions) indicated by conventional sequencing. Genome-scale methods uniquely enable the calculation of clone fitness values by providing concentration data for all clones within a genomic library before and after a period of selection. We observed that clone fitness values differ substantially from clone concentration values and that this is due to differences in overall clone fitness distributions for each selection. Finally, we show that many of the clones of highest fitness overlapped across all selections, suggesting that inhibition of aspartate metabolism, as opposed to specific inhibited enzymes, dominated each selection. Our follow up studies confirmed our observed growth phenotypes and showed that intracellular amino-acid levels were also altered in several of the identified clones. These results demonstrate that genome-scale methods, such as SCALEs, can be used to dramatically improve understanding of classic strain engineering approaches.

Duke Scholars

Published In

Metabolic engineering

DOI

EISSN

1096-7184

ISSN

1096-7176

Publication Date

March 2008

Volume

10

Issue

2

Start / End Page

109 / 120

Related Subject Headings

  • Selection, Genetic
  • Plasmids
  • Phenotype
  • Oligonucleotide Array Sequence Analysis
  • Nucleic Acid Hybridization
  • Mutagenesis, Insertional
  • Lac Operon
  • Genomics
  • Genomic Library
  • Genome, Bacterial
 

Citation

APA
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MLA
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Bonomo, J., Lynch, M. D., Warnecke, T., Price, J. V., & Gill, R. T. (2008). Genome-scale analysis of anti-metabolite directed strain engineering. Metabolic Engineering, 10(2), 109–120. https://doi.org/10.1016/j.ymben.2007.10.002
Bonomo, Jeanne, Michael D. Lynch, Tanya Warnecke, James V. Price, and Ryan T. Gill. “Genome-scale analysis of anti-metabolite directed strain engineering.Metabolic Engineering 10, no. 2 (March 2008): 109–20. https://doi.org/10.1016/j.ymben.2007.10.002.
Bonomo J, Lynch MD, Warnecke T, Price JV, Gill RT. Genome-scale analysis of anti-metabolite directed strain engineering. Metabolic engineering. 2008 Mar;10(2):109–20.
Bonomo, Jeanne, et al. “Genome-scale analysis of anti-metabolite directed strain engineering.Metabolic Engineering, vol. 10, no. 2, Mar. 2008, pp. 109–20. Epmc, doi:10.1016/j.ymben.2007.10.002.
Bonomo J, Lynch MD, Warnecke T, Price JV, Gill RT. Genome-scale analysis of anti-metabolite directed strain engineering. Metabolic engineering. 2008 Mar;10(2):109–120.
Journal cover image

Published In

Metabolic engineering

DOI

EISSN

1096-7184

ISSN

1096-7176

Publication Date

March 2008

Volume

10

Issue

2

Start / End Page

109 / 120

Related Subject Headings

  • Selection, Genetic
  • Plasmids
  • Phenotype
  • Oligonucleotide Array Sequence Analysis
  • Nucleic Acid Hybridization
  • Mutagenesis, Insertional
  • Lac Operon
  • Genomics
  • Genomic Library
  • Genome, Bacterial