Difference between revisions of "Part:BBa K2100005"
 
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<partinfo>BBa_K2100005 short</partinfo>
 
<partinfo>BBa_K2100005 short</partinfo>
  
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eYFP is a fluorescent protein constituitively expressed with the hEF1a (human elongation factor 1 alpha) promoter. The promoter and gene are encoded within an expression vector backbone with an SV40 terminator site after the eYFP gene and an ampicillin resistant gene.
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eYFP is a yellow fluorescent protein.
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2100005 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2100005 SequenceAndFeatures</partinfo>
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The MIT iGEM team used the hEF1a promoter to constitutively express fluorescent proteins as transfection markers in our experiments in HEK293, tHESC, MCF-7, and ISH cells. The amount of fluorescent units observed is an indication of the number of plasmids a particular cell uptakes. The graph below shows two single color controls from one of our experiments. We transfected one well of HEK293 cells with hEF1a-mKate and another with hEF1a-eYFP. The cells that uptook the hEF1a-mKate plasmids showed an increase in only the red fluorescent output, while the cells that uptook the hEF1a-eYFP plasmids showed a clear increase in only the yellow fluorescent output.
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https://static.igem.org/mediawiki/parts/c/c8/T--MIT--hEF1a.png
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Here is an image of HEK293 cells transfected with the eYFP florescent gene cascaded with a hEF1a constituitive promoter. We often used yellow florescence as either a reporter for how our promoter responded to induction in cells or as transfection markers for experiments.
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https://static.igem.org/mediawiki/parts/thumb/4/49/T--MIT--khb_eYFPexpression_hek.jpeg/796px-T--MIT--khb_eYFPexpression_hek.jpeg
  
  

Latest revision as of 23:07, 28 October 2016


pENTR eYFP

eYFP is a yellow fluorescent protein.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 222
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 222
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 222
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 222
    Illegal AgeI site found at 5
  • 1000
    COMPATIBLE WITH RFC[1000]

The MIT iGEM team used the hEF1a promoter to constitutively express fluorescent proteins as transfection markers in our experiments in HEK293, tHESC, MCF-7, and ISH cells. The amount of fluorescent units observed is an indication of the number of plasmids a particular cell uptakes. The graph below shows two single color controls from one of our experiments. We transfected one well of HEK293 cells with hEF1a-mKate and another with hEF1a-eYFP. The cells that uptook the hEF1a-mKate plasmids showed an increase in only the red fluorescent output, while the cells that uptook the hEF1a-eYFP plasmids showed a clear increase in only the yellow fluorescent output.

T--MIT--hEF1a.png

Here is an image of HEK293 cells transfected with the eYFP florescent gene cascaded with a hEF1a constituitive promoter. We often used yellow florescence as either a reporter for how our promoter responded to induction in cells or as transfection markers for experiments.

796px-T--MIT--khb_eYFPexpression_hek.jpeg