Part:BBa_K3939111

Fast degrading GFP

The ubiquitin tag is fused to the N-terminal of the GFP to make the GFP a rapidly degrading GFP

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 277
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 25
  Illegal AgeI site found at 466
  
• 1000
  COMPATIBLE WITH RFC[1000]

We improved the GFP part. In the beginning, we directly took the GFP part into practice, while later, it is observed that GFP has a relatively long half-life, which is not conducive to further measurement. This section we will illustrate how we test our improvel designs correspondingly, showing the reasonability in the process and the reliability of the results.<p>

Measurement of nfGFP half-life

1. Characterization part design:

<p>Ubiquitin is a small protein found in all eukaryotes whose primary function is to label a protein to be quickly recognized and degraded by the proteasome in the cell. We added a ubiquitin tag before the GFP structural gene, predicting that this would result in faster recognition and degradation of the expressed ubiquitin-tagged-GFP fusion protein by the ubiquitin degradation mechanism. To test our hypothesis, we designed the following parts and experiments.

Fig.1 nfGFP Schematic (plasmid vector 、Pgal、Ubiquitin tag、nfGFP、terminator)