Difference between revisions of "Part:BBa J100255:Experience"
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This device is been leaky and allows production of the tetA protein in the absence of theophylline which makes the cells tetracycline resistant in the absence of added theophylline. Furthermore, when theophylline is added, the cells did not grow well +/- tetracycline which indicates too much tetA protein is produced (lethality). The data below are typical of what we got on multiple experiments. <br><Br> | This device is been leaky and allows production of the tetA protein in the absence of theophylline which makes the cells tetracycline resistant in the absence of added theophylline. Furthermore, when theophylline is added, the cells did not grow well +/- tetracycline which indicates too much tetA protein is produced (lethality). The data below are typical of what we got on multiple experiments. <br><Br> | ||
| − | We strongly recommend people use [[Part:BBa_J100279]] which works much better. ''J100279'' uses weaker P9 promoter and weaker BD10 RBS. All of these were tested on pSB1A2 high copy plasmid in JM109 cells. | + | We strongly recommend people use [[Part:BBa_J100279]] which works much better. '''J100279''' uses weaker P9 promoter and weaker BD10 RBS. All of these were tested on pSB1A2 high copy plasmid in JM109 cells. |
<br> | <br> | ||
<center> | <center> | ||
| − | [[File:tClone_Tet_original.png]] | + | [[File:tClone_Tet_original.png|700px|]]<br> |
| − | Data collected from cells grown and measured at the same time. The major problems are too high cell growth in column 2, and too little cell growth in columns 3 and 4. This combination of P5 and BD18 should be used for avoided due to excessive leaking. Convert to ''J100279'' instead | + | Data collected from cells grown and measured at the same time. The major problems are too high cell growth in column 2, and too little cell growth in columns 3 and 4. This combination of P5 and BD18 should be used for avoided due to excessive leaking. Convert to '''J100279''' instead. |
</center> | </center> | ||
Latest revision as of 15:35, 8 July 2016
This device is been leaky and allows production of the tetA protein in the absence of theophylline which makes the cells tetracycline resistant in the absence of added theophylline. Furthermore, when theophylline is added, the cells did not grow well +/- tetracycline which indicates too much tetA protein is produced (lethality). The data below are typical of what we got on multiple experiments.
We strongly recommend people use Part:BBa_J100279 which works much better. J100279 uses weaker P9 promoter and weaker BD10 RBS. All of these were tested on pSB1A2 high copy plasmid in JM109 cells.
Data collected from cells grown and measured at the same time. The major problems are too high cell growth in column 2, and too little cell growth in columns 3 and 4. This combination of P5 and BD18 should be used for avoided due to excessive leaking. Convert to J100279 instead.
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