Part:BBa_K2751010

Ubc9-YPet

Usage and Biology

The relations between ubiquitin and SUMO has been widely studied. There is also research that applies the FRET technique in observing the combination of Ubc 9 and SUMO1.

To assure that we can conduct FRET experiments and to check the qualities of our protein, we closely mimic the experiments in a research[1]. In this process, we constructed a FRET pair of YPet-Ubc9 and CyPet-SUMO1. The Ubc9 and SUMO1 of the fusion proteins are expected to bind to each other, pulling YPet CyPet close enough to activate FRET reaction.

We have successfully conducted this part of experiment and the results are shown below. The part has been proved functional and the results can act as a positive control for our FRET model.

Reference: 1. Song, Y., & Liao, J. (2012, August). An in vitro Förster resonance energy transfer-based high-throughput screening assay for inhibitors of protein-protein interactions in SUMOylation pathway. Retrieved from https://www.ncbi.nlm.nih.gov/pubmed/22192309

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 40
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 529
  Illegal XhoI site found at 1258
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1181