Chemically inducible chromosomal evolution (CIChE) for multicopy metabolic pathway engineering

Aaron M. Love, Bradley W. Biggs, Keith Edward Jaggard Tyo, Parayil Kumaran Ajikumar*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Chemically inducible chromosomal evolution (CIChE) was developed for stable multicopy chromosomal integration of heterologous genes. In this technique, flanking an antibiotic selection marker and a gene of interest with identical regions of homology permits gene duplication via recA mediated homologous recombination. A strong selective pressure for gene duplication can be applied by increasing antibiotic concentration, and in a week’s time one can create a set of strains with a wide range of cassette copy numbers (upward of 20×), which can be made stable by deletion of recA. Herein, we describe a generalized workflow for this methodology.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc
Pages37-45
Number of pages9
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1927
ISSN (Print)1064-3745

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Keywords

  • Chromosomal integration
  • Escherichia coli
  • Metabolic engineering
  • Molecular cloning
  • Synthetic biology
  • Tools

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Love, A. M., Biggs, B. W., Tyo, K. E. J., & Ajikumar, P. K. (2019). Chemically inducible chromosomal evolution (CIChE) for multicopy metabolic pathway engineering. In Methods in Molecular Biology (pp. 37-45). (Methods in Molecular Biology; Vol. 1927). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9142-6_4