Difference between revisions of "Part:BBa K613015"
| Line 8: | Line 8: | ||
This TetR mutant was characterized ''in vivo'' by putting it into [https://parts.igem.org/Part:pSB3K1 pSB3K1] under a constitutive promoter ([https://parts.igem.org/Part:J23116 J23116]). This plasmid was cotransformed with [https://parts.igem.org/Part:J61002 J61002] harbouring RFP under pTet promoter ([https://parts.igem.org/Part:B0040 B0040]) in DH5alpha cells. Cells were grown in a medium containing Kanamycin & Amplicillin plus different concentrations of ATC, ranging from 0 to 2000 ng/mL. OD600 absorbence and RFP fluorescence were measured every 10 minutes during 12 hours on a platereader machine. | This TetR mutant was characterized ''in vivo'' by putting it into [https://parts.igem.org/Part:pSB3K1 pSB3K1] under a constitutive promoter ([https://parts.igem.org/Part:J23116 J23116]). This plasmid was cotransformed with [https://parts.igem.org/Part:J61002 J61002] harbouring RFP under pTet promoter ([https://parts.igem.org/Part:B0040 B0040]) in DH5alpha cells. Cells were grown in a medium containing Kanamycin & Amplicillin plus different concentrations of ATC, ranging from 0 to 2000 ng/mL. OD600 absorbence and RFP fluorescence were measured every 10 minutes during 12 hours on a platereader machine. | ||
| + | |||
'''Induction curves''' | '''Induction curves''' | ||
Fluorescence measurements (RFUs) were normalized by OD600 values. | Fluorescence measurements (RFUs) were normalized by OD600 values. | ||
| + | |||
| + | [[Image:EPFL_TetR-E37AW43ST141A-induction.png|600px]] | ||
| + | |||
'''Dose-response curve''' | '''Dose-response curve''' | ||
Fluorescence measurements (RFUs) were normalized by OD600 values. For each ATC concentration, we estimated the steady-state fluorescence expression by averaging the measurements over the last hour. | Fluorescence measurements (RFUs) were normalized by OD600 values. For each ATC concentration, we estimated the steady-state fluorescence expression by averaging the measurements over the last hour. | ||
| + | |||
| + | [[File:EPFL_TetR-E37AW43ST141A-doseresponse.png|600px]] | ||
| + | |||
<!-- --> | <!-- --> | ||
Revision as of 20:45, 21 September 2011
TetR E37A W43S T141A mutant
This is a TetR mutant, who carries the triple mutation EA37WS43TA141. Is part of the muTetR toolkit.
In vivo characterization
This TetR mutant was characterized in vivo by putting it into pSB3K1 under a constitutive promoter (J23116). This plasmid was cotransformed with J61002 harbouring RFP under pTet promoter (B0040) in DH5alpha cells. Cells were grown in a medium containing Kanamycin & Amplicillin plus different concentrations of ATC, ranging from 0 to 2000 ng/mL. OD600 absorbence and RFP fluorescence were measured every 10 minutes during 12 hours on a platereader machine.
Induction curves
Fluorescence measurements (RFUs) were normalized by OD600 values.
Dose-response curve
Fluorescence measurements (RFUs) were normalized by OD600 values. For each ATC concentration, we estimated the steady-state fluorescence expression by averaging the measurements over the last hour.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]