Difference between revisions of "Part:BBa K1061013"
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===Usage and Biology===
 
===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1061013 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1061013 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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The 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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Revision as of 18:21, 26 September 2013

P tight promoter

It is a response element containing 7 consecutive TetO elements and a minimal CMV promoter. When binding to tTA or rtTA, the expression of the downstream gene will be highly activated. Besides, it can achieve low constitutive expression(almost undectectable) when not binding to tTA or rtTA.

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

The 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