Coding

Part:BBa_K3794004

Designed by: Venkata Kalyan Ghadiyaram, Harshraj Bumia   Group: iGEM21_KCL_UK   (2021-09-30)
Revision as of 19:41, 19 October 2021 by Harshbumia (Talk | contribs)


ChABC CDS

Chondroitinase ABC (ChABC) is a bacterial enzyme secreted by Proteus vulgaris , it degrades the glycosaminoglycan (GAG) chains of chondroitin sulfate proteo-glycans (CSPGs) to allow for axonal regeneration following spinal cord injuries.

This ChABC part is derived from PDB ID: 1HN0, and also encodes a 6xHis tag and a TEV cleavage site for protein purification via a Ni-NTA column.

ChABC can be administered in the form of microinjections to the glial scar that is formed during a spinal cord injury, to degrade GAG chains from CSPGs - which are inhibitory molecules to axonal regeneration.

Due to the thermal instability of native ChABC at physiological temperatures, 8 point mutations to confer thermostability at 37C were introduced before synthesis of part in its composite form by IDT.

These mutations are as follows:

  • K194E
  • A228K
  • S274P
  • N288D
  • S343N
  • K654D
  • R670T
  • Q781E

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2754
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None