Difference between revisions of "Part:BBa K613016"

 
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<partinfo>BBa_K613016 short</partinfo>
 
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This is a TetR mutant who carries the P39K mutation.
 
This is a TetR mutant who carries the P39K mutation.
  
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===Usage and Biology===
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===In vivo characterization===
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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.
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'''Induction curves'''
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Fluorescence measurements (RFUs) were normalized by OD600 values.
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'''Dose-response curve'''
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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.
  
 
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Revision as of 17:56, 21 September 2011

TetR P39K mutant

This is a TetR mutant who carries the P39K mutation.


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]