Part:BBa_K898890:Design

wtPUF+PIN

Assembly Compatibility:

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

  Illegal XhoI site found at 1297
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 35
  
• 1000
  COMPATIBLE WITH RFC[1000]

Characterization Method

This figure includes the resulting PUF-PIN proteins interacting with the binding sites. The wild type binding site placed in between RBS and YFP, is represented by the orange oval. The yellow rounded rectangle represents the YFP reporter gene.

With regards to our hypothetical results, our experimental fluorescence measurements were substantially faithful to our predictions. Looking at columns 5, 6, 7, and 8, our observations appear to be in line with what we have predicted in introducing the wild type PUF-PIN to constructs containing a control recognition site and a specific recognition site. Columns 1 and 2 are negative fluorescence controls while Columns 3 and 4 are positive fluorescence controls measured without the influence of any sort of binding site.

Wild type PUF-PIN fusion bound and cleaved RNA as expected.

Source

The PUF domain from human Pum1 (residue 828-1176) and the PIN domain from human Smg6 (residue 1238-1421)

References

Professor Zefeng Wang, University of North Carolina- Chapel Hill

http://www.med.unc.edu/pharm/people/primaryfaculty/zefeng-wang/wang