Overview on the Expression of Toxic Gene Products in Escherichia coli

Fakhri Saïda1

1 University of California San Diego, La Jolla, California
Publication Name:  Current Protocols in Protein Science
Unit Number:  Unit 5.19
DOI:  10.1002/0471140864.ps0519s50
Online Posting Date:  November, 2007
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Despite the development of various nonbacterial expression systems, Escherichia coli (E. coli) remains the host of choice for recombinant protein expression. Its culture is simple, fast, inexpensive, and highly efficient (tens of milligrams of pure proteins are typically obtained within 48 hours using as little as 1 liter of culture). Unfortunately, many toxic genes (from various organisms) severely interfere with the physiology of E. coli. As a result, expression yields are dramatically diminished, and sometimes abolished. In fact, some genes are so toxic that E. coli cannot maintain their expression vector during the growth phase (the phase during which recombinant gene expression is presumably repressed). Therefore, modified expression vectors, modified E. coli strains, and appropriate cultivation protocols are needed. This overview discusses several special strategies successfully used to express toxic genes in E. coli. Curr. Protoc. Protein Sci. 50:5.19.1‐5.19.13. © 2007 by John Wiley & Sons, Inc.

Keywords: toxic genes; expression; E. coli

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Table of Contents

  • Introduction
  • Expression Protocols That Do Not Require Host or Vector Modification
  • Experimental Selection of New E. coli Strains That Tolerate Toxic Genes
  • Trans Delivery of T7 RNAP Using Bacteriophage Derivatives
  • Modification of the Promoter Region
  • Addition of Transcription Terminators
  • Modification of the Coding Sequence
  • Control of the Copy Number
  • Addition of Cell Death Modules
  • Conclusion
  • Literature Cited
  • Figures
  • Tables
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