Part:BBa_K3794004

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. Upstream of the 6xHis tag, there is a start codon to allow for transcriptional initiation.

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 (BBa_K3794005) 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]