Explanatory chapter: Troubleshooting recombinant protein expression: General

Krisna C. Duong-Ly, Sandra B. Gabelli

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

One of the most daunting problems for biochemists is the expression of recombinant proteins. Often, the host organism differs from the organism from which the gene coding for the protein of interest was derived. This article provides guidelines to determine whether or not protein expression is a problem, describes possible reasons for low protein expression, and covers several possible solutions. A protocol for measuring protein expression during E. coli cell growth and after induction is given. The reader should note that low protein expression is a complex problem that often stems from a variety of factors. Combinations of the solutions presented in this article may be required to solve a problem of protein expression. A brief overview of host cell expression systems is given, but the article primarily focuses on expression in E. coli as this is the most commonly used host organism. Some of the methods discussed here, however, may be applied to other expression systems.

Original languageEnglish (US)
Title of host publicationLaboratory Methods in Enzymology
Subtitle of host publicationProtein Part C
PublisherAcademic Press Inc.
Pages209-229
Number of pages21
ISBN (Print)9780124201194
DOIs
StatePublished - 2014

Publication series

NameMethods in Enzymology
Volume541
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • E. coli cell growth
  • Eukaryotic expression systems
  • Miniprep DNA
  • T7 RNA polymerase
  • T7 expression vectors
  • XL1-Blue cells

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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