Tetracycline: Difference between revisions
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*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. | ||
*Stock solution is | *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. | ||
==Plate Color Code== | ==Plate Color Code== | ||
Revision as of 12:38, 19 December 2005
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. Add 1 ml per liter of cooled broth or agar. Solutions and plates are light sensitive.
Plate Color Code
Yellow stripe
Properties
- Molecular Weight - 480.9g
