Difference between revisions of "Part:BBa K3610031"
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After successful transformation of yeast cells we checked for expression of the protein under a confocal microscope. Expression of the construct was confirmed. We failed, however, to confirm localization at the cell membrane.
 
After successful transformation of yeast cells we checked for expression of the protein under a confocal microscope. Expression of the construct was confirmed. We failed, however, to confirm localization at the cell membrane.
  
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[[File:T--UZurich--BAK1(-).png|800px|thumb|left|]]
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[[File:T--UZurich--BAK1(-).png|800px|]]
  
  

Revision as of 16:22, 18 October 2020


BAK1 without native signal peptide / YFP

This part can be used for expressing the plant pattern recognition receptor BRI1-associated receptro kinase (BAK1) from Arabidopsis thaliana in S. cerevisiae.

The sequence of the BAK1 protein does not contain the original signal peptide sequence of the receptor (BAK-). Instead it has been replaced by the secretion signal of the alpha-Factor from S. cerevisiae. To make expression of BAK1 visible and to test observe the localization in the cell, the BAK1 coding region has been fused to a yellow fluorescent protein by a 15 amino-acid long linker.

For initiation of the expression the promoter of this part is a truncated version of the ADH1 promoter from S. cerevisiae. For termination this part has terminator sequence of the enolase 2 protein from S. cerevisiae.

Usage and Biology

Characterization

Expression of BAK1-/YFP in S. cerevisiae

After successful transformation of yeast cells we checked for expression of the protein under a confocal microscope. Expression of the construct was confirmed. We failed, however, to confirm localization at the cell membrane.

T--UZurich--BAK1(-).png

















Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1758
    Illegal PstI site found at 848
    Illegal PstI site found at 893
    Illegal PstI site found at 1183
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1758
    Illegal PstI site found at 848
    Illegal PstI site found at 893
    Illegal PstI site found at 1183
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1758
    Illegal PstI site found at 848
    Illegal PstI site found at 893
    Illegal PstI site found at 1183
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1758
    Illegal PstI site found at 848
    Illegal PstI site found at 893
    Illegal PstI site found at 1183
  • 1000
    COMPATIBLE WITH RFC[1000]