Difference between revisions of "Part:BBa K613013"
(→In vivo characterization) |
|||
| Line 11: | Line 11: | ||
===In vivo characterization=== | ===In vivo characterization=== | ||
| − | 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 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 and RFP fluorescence were measured every 10 minutes during 12 hours on a platereader machine. |
'''Induction curves''' | '''Induction curves''' | ||
Revision as of 17:43, 21 September 2011
TetR V36F mutant
TetR with a point mutation V36F.
In vitro characterization
à toi lilia :)
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 and RFP fluorescence were measured every 10 minutes during 12 hours on a platereader machine.
Induction curves
The RFUs have been normalized by OD600 to have a better comparison between the different ATC concentrations.
Dose-response curve
For each ATC concentration, RFUs have been first divided by the corresponding OD600 value. Then, the last 6 values have been averaged to get the plateau value.
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]