Generator

Part:BBa_K2068007:Design

Designed by: Nithin Dharmaraj   Group: iGEM16_UCLA   (2016-09-13)


O3-33 Mutant aa118


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 679
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 354
    Illegal AgeI site found at 283
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Due to the unpredictable nature of protein folding especially of a self assembling protein cage a few criterion had to be met for mutant insertion into gene. 1. Avoid regions with secondary structure. This would theoretically lessen the chance of the mutation disrupting cage formation and allow the cage to retain wild type structure.Using the PDB file of our mutant wild type cage we were able to identify areas of the genome that had minimal to no secondary structure. 2. Make sure the mutation is on the outside of the cage. Using Pymol modeling we were able to see what regions of the genome coded for the residues on the surface of the cage and what regions coded for residues on the interior of the cage. Because we are introducing a protease to cleave our cage, we needed the cleavage site to be as sterically unhindered as possible.



Source

This is an mutated version of our wildtype protein cage that was initially designed by Dr. Neil King from University of Washington.

References