Difference between revisions of "Part:BBa K117004"
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__NOTOC__
 
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<partinfo>BBa_K117004 short</partinfo> <br>
 
<partinfo>BBa_K117004 short</partinfo> <br>
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This biobrick device has a lacI regulated promoter which upon the induction of lactose/IPTG, will casue the expression of the reporter gene, Green fluorescence protein (GFP EE0040).  
 
This biobrick device has a lacI regulated promoter which upon the induction of lactose/IPTG, will casue the expression of the reporter gene, Green fluorescence protein (GFP EE0040).  
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[[Image:NTU LacI GFP.png|thumb|center|800px| ''‎Function of K117004'' ]]
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==Abnormality found in the registry==
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This device could be found in the registry in the form of [https://parts.igem.org/wiki/index.php?title=Part:BBa_J04430 BBa_J04430]. However, the team found an abnormality when BBa_J04430 was transformed into Top 10 cells. As shown on the pictures below, the cells glow pink instead of green under UV light.
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[[image:NTU@iGEM_24062008_(pink_gfp).JPG|thumb|center|300px| BBa_J04430 glows pink]]
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==Making our own part==
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Hence the team constructed this part from the registry by ligating [https://parts.igem.org/wiki/index.php?title=Part:BBa_R0010 BBa_R0010] with [https://parts.igem.org/Part:BBa_E0240 BBa_E02040]. The BBa_E02040 consist of the biobrick parts: BB0034, E0040, B0010 and B0012.
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==Characterization of pLacI-GFP==
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The NTU@iGEM team characterized the biobrick part pLacI-GFP. The influence of 1)concentration of Lactose 2) temperature and 3) time were investigated.
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Shown below is the graph obtained for the experiment conducted at 37°C.
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[[image:NTU@iGEM_37norm.jpg|thumb|center|400px| Characterization at 37°C]]
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===Electron microscope pictures===
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[[image:NTU@iGEM_Fluorscence_LacI-GFP_1mM.JPG|thumb|center|300px| Cells induced with 1mM lactose concentration]]
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[[image:NTU@iGEM_Fluorscence_LacI-GFP_10mM.JPG|thumb|center|300px| Cells induced with 10mM lactose concentration]]
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For more information including the protocols and results obtained, please go to:
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[http://2008.igem.org/Team:NTU-Singapore/Parts/Characterization_of_LacI-GFP NTU@iGEM Characterization of pLac-GFP]
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 12:58, 27 October 2008

pLacI-GFP

The construct design

‎BBa K117004

This biobrick device has a lacI regulated promoter which upon the induction of lactose/IPTG, will casue the expression of the reporter gene, Green fluorescence protein (GFP EE0040).

‎Function of K117004

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 873