Difference between revisions of "Part:BBa K1054022"

 
 
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<partinfo>BBa_K1054022 short</partinfo>
 
<partinfo>BBa_K1054022 short</partinfo>
  
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pLAC promotor,theophylline-inducible Riboswitch(helix-spin), GFP+Terminator
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This part is used to standardize the theophylline riboswitch with a helix spin mechanism. It acts differently from another throphylline inducible riboswitch (BBa_K411003).
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The protocols of characterization is as follows:
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1. Cultivate the BL21 containing BBa_ in LB at 37℃180rpm until OD600 reaches 0.5.
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2. Induce the bacteria with 1mM IPTG (final concentration), and cultivate them at 37℃180rpm for 4hrs.
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3. Dilute the suspension culture with LB to 0.1OD600 . Theophylline is added at concentration ranging from 1.00E-06 mol/L to1.00E-01 mol/L.
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4. Measure the RFU ( ex488/em525) and OD600 synchronously.
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5. Data Analysis
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[[File:500px-ZJU-Standardization 3.1.jpg]]
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Graphic Model about how RFU/OD changes at different time and different concentration of Theophylline.
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[[File:500px-ZJU-Standardization 3.2.jpg]]
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Graphic Model about how GFP Change Rate changes at different time and different concentration of Theophylline.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 22:40, 27 September 2013

pLAC promotor,theophylline-inducible Riboswitch(helix-spin), GFP+Terminator

This part is used to standardize the theophylline riboswitch with a helix spin mechanism. It acts differently from another throphylline inducible riboswitch (BBa_K411003).

The protocols of characterization is as follows:

1. Cultivate the BL21 containing BBa_ in LB at 37℃180rpm until OD600 reaches 0.5.

2. Induce the bacteria with 1mM IPTG (final concentration), and cultivate them at 37℃180rpm for 4hrs.

3. Dilute the suspension culture with LB to 0.1OD600 . Theophylline is added at concentration ranging from 1.00E-06 mol/L to1.00E-01 mol/L.

4. Measure the RFU ( ex488/em525) and OD600 synchronously.

5. Data Analysis


500px-ZJU-Standardization 3.1.jpg

Graphic Model about how RFU/OD changes at different time and different concentration of Theophylline.


500px-ZJU-Standardization 3.2.jpg

Graphic Model about how GFP Change Rate changes at different time and different concentration of Theophylline.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 969