Tetracycline

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==Resistance Mechanism==
==Resistance Mechanism==
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There are a number of resistance mechanisms, most require tetracycline antiporters.  The most commonly used is the tetA(C) gene.  The gene product of tetA(C) forms a pore in the inner membrane and exports one tet molecule for every proton imported.  [[Molecular Cloning]] states that tet resistance has a number of adverse affects on the cell, chief among those being a reduction in growth rate and viability.
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There are a number of resistance mechanisms, most require tetracycline antiporters.  The most commonly used is the tetA(C) gene.  The gene product of tetA(C) forms a pore in the inner membrane and exports one tet molecule for every proton imported.  [[Molecular Cloning]] states that tet resistance has a number of adverse affects on the cell, chief among those being a reduction in growth rate and viability.  
==Working concentration and Stock Solutions==
==Working concentration and Stock Solutions==
*Using a plasmid with copy number 15-20, 20μg/ml tet has proved a successful selector.  The reduction in growth rate, described above was significant.
*Using a plasmid with copy number 15-20, 20μg/ml tet has proved a successful selector.  The reduction in growth rate, described above was significant.
*The RajBhandary lab uses 5μg/ml and see a minimal reduction in growth rate and never lose plasmid from their cells.
*The RajBhandary lab uses 5μg/ml and see a minimal reduction in growth rate and never lose plasmid from their cells.
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*Stock solution is 15 mg/ml in 50% ethanol.  Add 1 ml per liter of cooled broth or agar.  Solutions and plates are light sensitive.
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*Stock solution is 15 mg/ml in 50% ethanol (note: using 95-100% ethanol is much better for dissolving stock powder).  Add 1 ml per liter of cooled broth or agar.  Solutions and plates are light sensitive.
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*Tetracycline is incompatible with (chelated by) magnesium ions, and thus unsuitable for use in minimal media
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==Plate Color Code==
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==Counterselection techniques==
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Yellow stripe
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*Fusaric acid can be used to select against tetracycline resistant colonies.  See PMID 6259126, and modifications PMID 7007341.
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*Nickel ions can also be used for tetR counterselection.  See PMID 8954882.
==Properties==
==Properties==
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*'''Molecular Weight''' - 480.9g
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*'''Molecular Weight''' - 480.9 g/mol (Hydrochloride salt), 444.4 g/mol (free base)
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** (This value doesn't agree with the number given in the Wikipedia link below)
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==References==
==References==
*[[Molecular Cloning]]
*[[Molecular Cloning]]
*[[Wikipedia:Tetracycline|Wikipedia Tetracycline page]]
*[[Wikipedia:Tetracycline|Wikipedia Tetracycline page]]
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[[Category:Material]] [[Category:Antibiotic]]

Current revision

Contents

Mode of Action

Tetracyclines (there are many) enter the cell by diffusion through OmpF porins. It inhibits growth by preventing codon-anticodon interactions during translation.

Resistance Mechanism

There are a number of resistance mechanisms, most require tetracycline antiporters. The most commonly used is the tetA(C) gene. The gene product of tetA(C) forms a pore in the inner membrane and exports one tet molecule for every proton imported. Molecular Cloning states that tet resistance has a number of adverse affects on the cell, chief among those being a reduction in growth rate and viability.

Working concentration and Stock Solutions

  • Using a plasmid with copy number 15-20, 20μg/ml tet has proved a successful selector. The reduction in growth rate, described above was significant.
  • The RajBhandary lab uses 5μg/ml and see a minimal reduction in growth rate and never lose plasmid from their cells.
  • Stock solution is 15 mg/ml in 50% ethanol (note: using 95-100% ethanol is much better for dissolving stock powder). Add 1 ml per liter of cooled broth or agar. Solutions and plates are light sensitive.
  • Tetracycline is incompatible with (chelated by) magnesium ions, and thus unsuitable for use in minimal media

Counterselection techniques

  • Fusaric acid can be used to select against tetracycline resistant colonies. See PMID 6259126, and modifications PMID 7007341.
  • Nickel ions can also be used for tetR counterselection. See PMID 8954882.

Properties

  • Molecular Weight - 480.9 g/mol (Hydrochloride salt), 444.4 g/mol (free base)

References

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