Gene Dosage Experiments in Enterobacteriaceae Using Arabinose-regulated Promoters

Sanchari Bhattacharyya; Shimon Bershtein; Eugene I Shakhnovich

doi:10.21769/bioprotoc.2396

Gene Dosage Experiments in Enterobacteriaceae Using Arabinose-regulated Promoters

Sanchari Bhattacharyya, Shimon Bershtein, Eugene I Shakhnovich

Department of Life Sciences

Ben-Gurion University of the Negev

Research output: Contribution to journal › Article › peer-review

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.

Original language	American English
Article number	e2396
Journal	BIO-PROTOCOL
Volume	7
Issue number	14
DOIs	https://doi.org/10.21769/bioprotoc.2396
State	Published - Jun 2017

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10.21769/bioprotoc.2396

Cite this

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title = "Gene Dosage Experiments in Enterobacteriaceae Using Arabinose-regulated Promoters",

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.",

author = "Sanchari Bhattacharyya and Shimon Bershtein and Shakhnovich, {Eugene I}",

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AB - 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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