Difference between revisions of "Part:BBa K2520011"

 
 
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<partinfo>BBa_K2520011 short</partinfo>
 
<partinfo>BBa_K2520011 short</partinfo>
  
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EF1a-GFP-hGH
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This part is an improved part of <partinfo>K1119008</partinfo>. This device allows constitutive expression in mammalian cells of GFP. EF1a promoter is very useful in cells where other promoters, such as CMV, are underactive or silenced (such as stem cells).
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===Results===
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In order to test our mutant promoter, we tested the expression of the reporter gene GFP under three different promoters- CMV, EF1a WT and the mutant EF1a we created. As shown in the following graph, the expression levels of the reporter gene under EF1a promoter was much higher as compared to the CMV promoter, and these levels are almost equal between the WT and the mutant promoter.
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[[File:EF1a results new.jpeg|600px|thumb|center|Figure 1: Fold change of Fluorescence intensity of GFP under three different promoters- CMV, WT EF1a and mutant EF1a, compared with non-transfected cells, using Flow Cytometry, FITC channel. ]]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 12:36, 29 October 2017


EF1a-GFP-hGH

This part is an improved part of BBa_K1119008. This device allows constitutive expression in mammalian cells of GFP. EF1a promoter is very useful in cells where other promoters, such as CMV, are underactive or silenced (such as stem cells).

Results

In order to test our mutant promoter, we tested the expression of the reporter gene GFP under three different promoters- CMV, EF1a WT and the mutant EF1a we created. As shown in the following graph, the expression levels of the reporter gene under EF1a promoter was much higher as compared to the CMV promoter, and these levels are almost equal between the WT and the mutant promoter.

Figure 1: Fold change of Fluorescence intensity of GFP under three different promoters- CMV, WT EF1a and mutant EF1a, compared with non-transfected cells, using Flow Cytometry, FITC channel.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 643
    Illegal XhoI site found at 1042
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 777
    Illegal AgeI site found at 151
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2375
    Illegal BsaI.rc site found at 1904