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Gene Dosage Experiments in Enterobacteriaceae Using Arabinose-regulated Promoters

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2017

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Bhattacharyya, Sanchari, Shimon Bershtein, and Eugene I Shakhnovich. 2017. “Gene Dosage Experiments in Enterobacteriaceae Using Arabinose-regulated Promoters.” Bio-protocol 7 (14): e2396. doi:10.21769/BioProtoc.2396. http://dx.doi.org/10.21769/BioProtoc.2396.

Abstract

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.

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Gene-dosage toxicity, Arabinose, Protein abundance, Western-blot, Fitness, Over-expression, Plasmid

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