Difference between revisions of "Part:BBa K1061012"

 
 
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<partinfo>BBa_K1061012 short</partinfo>
 
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tTA is a fusion protein of tetR protein from E.coli and the transcription activation domain VP16 from simplex herpes virus. It can bind to the Ptight element in response to doxycyline, and activate the expression of gene downstream of Ptight element.
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tTA is a fusion protein of tetR protein from E.coli and the transcription activation domain VP16 from simplex herpes virus. It can bind to the Ptight element in response to doxycyline, and activate the expression of gene downstream of Ptight element.[1]
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1061012 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1061012 SequenceAndFeatures</partinfo>
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====Funtional experiment====
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We conducted the functional experiment by co-tranfected the plasmid contains tTA and the plasmid contains P tight into Bosc, one kind of HEK-293 cell lines.Noted that we did not use tTA advance due to time limit(we construct the plasmid contains tTA advance later), and we will try that later,for improvement of expression control.
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[[Image:P_tight_functional_test.jpg|thumb|center|800px|functional assay of tTA]]
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After adding dox, the expression of dox is almost fully suppressed.
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We also get the quantitive data by western blot, which will be submitted later.
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====quantitive analysis====
  
 
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<partinfo>BBa_K1061012 parameters</partinfo>
 
<partinfo>BBa_K1061012 parameters</partinfo>
 
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<h2>References:</h2>
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<p>[1]http://www.clontech.com/CN/Products/Inducible_Systems/Tetracycline-Inducible_Expression/Gen2?sitex=10022:22372:US

Latest revision as of 02:43, 29 September 2013

tTA

tTA is a fusion protein of tetR protein from E.coli and the transcription activation domain VP16 from simplex herpes virus. It can bind to the Ptight element in response to doxycyline, and activate the expression of gene downstream of Ptight element.[1]

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]

Funtional experiment

We conducted the functional experiment by co-tranfected the plasmid contains tTA and the plasmid contains P tight into Bosc, one kind of HEK-293 cell lines.Noted that we did not use tTA advance due to time limit(we construct the plasmid contains tTA advance later), and we will try that later,for improvement of expression control.

functional assay of tTA
After adding dox, the expression of dox is almost fully suppressed.

We also get the quantitive data by western blot, which will be submitted later.

quantitive analysis

References:

[1]http://www.clontech.com/CN/Products/Inducible_Systems/Tetracycline-Inducible_Expression/Gen2?sitex=10022:22372:US