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Related Concept Videos

Riboswitches01:56

Riboswitches

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Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
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Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...
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Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
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Types of RNA01:20

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Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in regulating gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
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Translational Regulation01:29

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Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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Global Regulatory Systems

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Global regulatory systems in bacteria enable rapid and coordinated responses to environmental changes by integrating sensory inputs with gene expression, ensuring efficient adaptation to fluctuating conditions. Key global regulatory mechanisms include regulons, two-component systems, sigma factors, and secondary messengers.Regulons and Global RegulatorsA regulon is a collection of genes and operons controlled by a common global regulator. These regulators enable bacteria to prioritize resource...
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Using riboswitches to regulate gene expression and define gene function in mycobacteria.

Erik R Van Vlack1, Jessica C Seeliger2

  • 1Department of Chemistry, Stony Brook University, Stony Brook, New York, USA.

Methods in Enzymology
|January 22, 2015
PubMed
Summary

We present methods for using theophylline-inducible riboswitches to control gene expression in mycobacteria. This technique aids in studying gene function and essentiality in various settings, including host cell infections.

Keywords:
Flow cytometryGFPHomologous recombinationInducibleInfectionMycobacteriaTheophyllineβ-Galactosidase

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Mycobacteria, including Mycobacterium tuberculosis, are significant human pathogens.
  • Controlling gene expression is crucial for understanding gene function and essentiality.
  • Theophylline-inducible artificial riboswitches offer a novel approach to gene regulation.

Purpose of the Study:

  • To provide detailed protocols for characterizing theophylline-inducible riboswitch performance in mycobacteria.
  • To enable the application of riboswitches for exploring gene function in both in vitro and host infection models.
  • To facilitate the use of riboswitches for studying essential genes in mycobacteria.

Main Methods:

  • Characterization of riboswitch response to theophylline in reporter strains.
  • Construction of recombinant mycobacteria with riboswitch-controlled endogenous genes.
  • Assessment of riboswitch functionality during host cell infections.

Main Results:

  • Established methods for evaluating riboswitch-mediated gene expression control.
  • Demonstrated theophylline-inducible gene expression in various mycobacterial systems.
  • Validated riboswitch application in reporter strains, endogenous gene regulation, and infection models.

Conclusions:

  • The described protocols effectively characterize theophylline-inducible riboswitches in mycobacteria.
  • This system provides a versatile tool for functional genomics in mycobacteria.
  • The methods support the exploration of gene function and essentiality in diverse mycobacterial research settings.