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Related Concept Videos

Operon Model01:23

Operon Model

The operon model represents a fundamental mechanism of gene regulation in prokaryotes, enabling coordinated expression of genes involved in related metabolic or functional pathways. Operons consist of structural genes, a promoter, and an operator, with transcription regulated by repressors, activators, and small effector molecules.Structure and Function of OperonsAn operon is a cluster of structural genes transcribed together under the control of a single promoter. The promoter region...
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...
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...
Operons02:09

Operons

Prokaryotes can control gene expression through operons—DNA sequences consisting of regulatory elements and clustered, functionally related protein-coding genes. Operons use a single promoter sequence to initiate transcription of a gene cluster (i.e., a group of structural genes) into a single mRNA molecule. The terminator sequence ends transcription. An operator sequence, located between the promoter and structural genes, prohibits the operon’s transcriptional activity if bound by a repressor...
Operons02:09

Operons

Prokaryotes can control gene expression through operons—DNA sequences consisting of regulatory elements and clustered, functionally related protein-coding genes. Operons use a single promoter sequence to initiate transcription of a gene cluster (i.e., a group of structural genes) into a single mRNA molecule. The terminator sequence ends transcription. An operator sequence, located between the promoter and structural genes, prohibits the operon’s transcriptional activity if bound by a repressor...
Repressible Operon: trp Operon01:21

Repressible Operon: trp Operon

The trp operon in Escherichia coli exemplifies a repressible operon. It regulates the synthesis of tryptophan through repressor-mediated transcriptional control and attenuation. This dual regulatory mechanism ensures tryptophan biosynthesis occurs only when needed, conserving cellular resources.Structure of the trp OperonThe trp operon consists of five structural genes (trpE, trpD, trpC, trpB, and trpA) that encode enzymes for tryptophan biosynthesis. These genes are transcribed as a single...

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Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX
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Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX

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Stoichiometric protein complex formation and over-expression using the prokaryotic native operon structure.

Christian Poulsen1, Simon Holton, Arie Geerlof

  • 1European Molecular Biology Laboratory (EMBL), Hamburg Outstation, c/o DESY, Hamburg, Germany.

FEBS Letters
|January 21, 2010
PubMed
Summary

This study shows that using native operon structures from mycobacteria in Mycobacterium smegmatis efficiently overexpresses protein complexes. This method works in both gram-positive and gram-negative bacteria for biochemical studies.

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Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX
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The Multifaceted Benefits of Protein Co-expression in Escherichia coli

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The MultiBac Protein Complex Production Platform at the EMBL
13:51

The MultiBac Protein Complex Production Platform at the EMBL

Published on: July 11, 2013

Area of Science:

  • Microbiology
  • Molecular Biology
  • Structural Biology

Background:

  • Prokaryotic operons frequently encode proteins that assemble into functional protein-protein complexes.
  • Efficient expression of these complexes is crucial for biochemical and structural studies.

Purpose of the Study:

  • To investigate the utility of native operon structures for overexpressing protein complexes.
  • To establish a robust method for producing prokaryotic protein complexes in a heterologous host.

Main Methods:

  • Utilized mycobacterial operons and Mycobacterium smegmatis as an expression host.
  • Demonstrated expression of protein complexes ranging from dimers to higher oligomers.
  • Tested the efficacy of the method across varying intergenic distances.
  • Extended the expression strategy to the gram-negative bacterium Escherichia coli.

Main Results:

  • Achieved robust and stoichiometrically correct expression of protein complexes.
  • Expression efficiency was largely unaffected by intergenic distances within operons.
  • Successfully transferred the expression strategy from a gram-positive (Mycobacterium) to a gram-negative (Escherichia coli) host.

Conclusions:

  • Native operon structures provide an efficient strategy for overexpressing prokaryotic protein complexes.
  • The method is versatile and functional across different bacterial species (gram-positive and gram-negative).
  • This approach offers a generalizable tool for producing large quantities of pure protein complexes for research.