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Regulated Expression Systems for Mycobacteria and Their Applications

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  • Authors: Dirk Schnappinger1, Sabine Ehrt3
  • Editors: Graham F. Hatfull5, William R. Jacobs Jr.6
  • VIEW AFFILIATIONS HIDE AFFILIATIONS
    Affiliations: 1: Department of Microbiology and Immunology, Weill Medical College of Cornell University, New York, NY 10065; 2: Program in Molecular Biology, Weill Graduate School of Medical Sciences of Cornell University, New York, NY 10065; 3: Department of Microbiology and Immunology, Weill Medical College of Cornell University, New York, NY 10065; 4: Program in Immunology and Microbial Pathogenesis, Weill Graduate School of Medical Sciences of Cornell University, New York, NY 10065; 5: University of Pittsburgh, Pittsburgh, PA 15260; 6: Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, NY 10461
    • Source: microbiolspec January 2014 vol. 2 no. 1 doi:10.1128/microbiolspec.MGM2-0018-2013
  • Received 14 August 2013 Accepted 27 August 2013 Published 17 January 2014
  • Dirk Schnappinger, [email protected] or Sabine Ehrt, [email protected]
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  • Abstract:

    For bacterial model organisms like and , genetic tools to experimentally manipulate the activity of individual genes have existed for decades. But for genetically less tractable yet medically important bacteria such as , such tools have rarely been available. More recently, several groups developed genetic switches that function efficiently in and other mycobacteria. Together these systems utilize six transcription factors, eight regulated promoters, and three regulatory principles. In this chapter we describe their design features, review their main applications, and discuss the advantages and disadvantages of regulating transcription, translation, or protein stability for controlling gene activities in bacteria.

    Genetic elements that enable specific and quantitative control over the activity of individual genes are irreplaceable components of the modern genetic toolbox. They facilitate not only the purification of proteins for biochemical, structural, or immunological studies but can also be applied to improve our understanding of gene functions. Until recently, only one such tool was available for use in mycobacteria, and its applicability in slowly growing mycobacteria was limited. But during the last decade at least a dozen new systems have been developed. In this chapter we review the design, components, and regulatory mechanisms of the different systems and discuss their main applications.

  • Citation: Schnappinger D, Ehrt S. 2014. Regulated Expression Systems for Mycobacteria and Their Applications. Microbiol Spectrum 2(1):MGM2-0018-2013. doi:10.1128/microbiolspec.MGM2-0018-2013.

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/content/journal/microbiolspec/10.1128/microbiolspec.MGM2-0018-2013
2014-01-17
2019-08-25

Abstract:

For bacterial model organisms like and , genetic tools to experimentally manipulate the activity of individual genes have existed for decades. But for genetically less tractable yet medically important bacteria such as , such tools have rarely been available. More recently, several groups developed genetic switches that function efficiently in and other mycobacteria. Together these systems utilize six transcription factors, eight regulated promoters, and three regulatory principles. In this chapter we describe their design features, review their main applications, and discuss the advantages and disadvantages of regulating transcription, translation, or protein stability for controlling gene activities in bacteria.

Genetic elements that enable specific and quantitative control over the activity of individual genes are irreplaceable components of the modern genetic toolbox. They facilitate not only the purification of proteins for biochemical, structural, or immunological studies but can also be applied to improve our understanding of gene functions. Until recently, only one such tool was available for use in mycobacteria, and its applicability in slowly growing mycobacteria was limited. But during the last decade at least a dozen new systems have been developed. In this chapter we review the design, components, and regulatory mechanisms of the different systems and discuss their main applications.

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Regulated Expression Systems for Mycobacteria and Their Applications
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Citation: Schnappinger D, Ehrt S. 2014. Regulated expression systems for mycobacteria and their applications. microbiolspec 2(1): doi:10.1128/microbiolspec.MGM2-0018-2013
/content/microbiolspec/10.1128/microbiolspec.MGM2-0018-2013.fig1
FIGURE 1
Regulatory systems for mycobacteria. The transcriptional regulatory systems are shown in to , the two controlled proteolysis systems in and , and the theophylline riboswitch in . Dotted lines ending in a perpendicular line indicate negative regulatory interactions; dotted lines ending in an arrow represent positive regulatory interactions. Ace, acetamide; tc/atc, tetracycline/anhydrotetracycline; IPTG, isopropyl β- 1-thiogalactopyranoside; ara, arabinose; IVN, isovaleronitrile; PI, pristinamycin.
microbiolspec/2/1/MGM2-0018-2013-fig1_thmb.gif
microbiolspec/2/1/MGM2-0018-2013-fig1.gif
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FIGURE 1

Regulatory systems for mycobacteria. The transcriptional regulatory systems are shown in to , the two controlled proteolysis systems in and , and the theophylline riboswitch in . Dotted lines ending in a perpendicular line indicate negative regulatory interactions; dotted lines ending in an arrow represent positive regulatory interactions. Ace, acetamide; tc/atc, tetracycline/anhydrotetracycline; IPTG, isopropyl β- 1-thiogalactopyranoside; ara, arabinose; IVN, isovaleronitrile; PI, pristinamycin.

Source: microbiolspec January 2014 vol. 2 no. 1 doi:10.1128/microbiolspec.MGM2-0018-2013
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Regulated Expression Systems for Mycobacteria and Their Applications
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Citation: Schnappinger D, Ehrt S. 2014. Regulated expression systems for mycobacteria and their applications. microbiolspec 2(1): doi:10.1128/microbiolspec.MGM2-0018-2013
/content/microbiolspec/10.1128/microbiolspec.MGM2-0018-2013.T1
TABLE 1
Regulated expression systems for mycobacteria and examples of their applications
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TABLE 1

Regulated expression systems for mycobacteria and examples of their applications

Source: microbiolspec January 2014 vol. 2 no. 1 doi:10.1128/microbiolspec.MGM2-0018-2013

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