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Related Concept Videos

Inducible Operons: lac Operon01:25

Inducible Operons: lac Operon

The lac operon in Escherichia coli is a model for understanding inducible gene regulation and metabolic flexibility. It integrates local control by lactose and global regulation through catabolite repression, enabling E. coli to preferentially metabolize glucose when available and switch to lactose utilization when glucose is scarce.Structure and Function of the lac OperonThe lac operon contains three structural genes: lacZ (β-galactosidase), lacY (lactose permease), and lacA (thiogalactoside...
Production of Pharmaceuticals01:30

Production of Pharmaceuticals

Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under sterile, tightly...
Reporter Genes02:11

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Prokaryotic Transcriptional Activators and Repressors01:58

Prokaryotic Transcriptional Activators and Repressors

The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
Transcription of prokaryotic...

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Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling
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Published on: June 25, 2015

Autoinduction of protein expression.

Brian G Fox1, Paul G Blommel1

  • 1Department of Biochemistry, Biophysics Degree Program, and Center for Eukaryotic Structural Genomics, University of Wisconsin-Madison, Madison, Wisconsin.

Current Protocols in Protein Science
|April 15, 2009
PubMed
Summary
This summary is machine-generated.

This study introduces simple protocols for lactose-derived autoinduction in Escherichia coli, enabling reproducible protein expression trials with minimal user effort for functional and structural studies.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Protein Expression

Background:

  • Escherichia coli is a widely used host for recombinant protein production.
  • Optimizing protein expression is crucial for functional and structural studies.
  • Autoinduction methods simplify the process of protein expression in bacterial systems.

Purpose of the Study:

  • To provide standardized protocols for lactose-derived autoinduction in Escherichia coli.
  • To enable reproducible protein expression with minimal user intervention.
  • To offer variations for different research needs, including labeling and high-throughput screening.

Main Methods:

  • Development of basic autoinduction protocols for unlabeled protein production.
  • Inclusion of protocols for selenomethionine labeling for X-ray crystallography.
  • Adaptation of protocols for multi-well plate formats for screening and optimization.

Main Results:

  • Demonstrated reproducibility in protein expression trials using the developed protocols.
  • Successful production of unlabeled proteins for functional studies.
  • Facilitation of selenomethionine labeling and multi-well plate screening.

Conclusions:

  • Lactose-derived autoinduction offers a robust and user-friendly method for Escherichia coli protein expression.
  • The provided protocols support diverse applications, from basic functional studies to advanced structural biology and screening.
  • These methods enhance efficiency and reproducibility in recombinant protein production.