Bio-Protocol (Jul 2017)

Gene Dosage Experiments in Enterobacteriaceae Using Arabinose-regulated Promoters

  • Sanchari Bhattacharyya,
  • Shimon Bershtein,
  • Eugene I Shakhnovich

DOI
https://doi.org/10.21769/bioprotoc.2396
Journal volume & issue
Vol. 7, no. 14

Abstract

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This protocol is used to assay the effect of protein over-expression on fitness of E. coli. It is based on a plasmid expression of a protein of interest from an arabinose-regulated pBAD promoter followed by the measurement of the intracellular protein abundance by Western blot along with the measurement of growth parameters of E. coli cell expressing this protein.