Part:BBa_K3833000

Streptococcal Collagen-Like Protein

Streptococcal Collagen-Like Protein (Scl2) was originally discovered in gram-positive S. pyogenes. It is a prokaryotic protein that can be used to mimic human collagen and has been shown to form stable triple helices[1].

Scl2 is easily expressed in E. coli and lacks the requirement for post-translational modifications; this makes it a useful alternative to recombinant human collagen expressed in eukaryotic organisms[1]. For these reasons, we chose Scl2 as the backbone for Collatrix — a collagen-mimetic gradient hydrogel for bone-soft tissue interfaces.

Scl2 can be modified to specific locations with common mammalian cell binding sites, such as fibronectin and integrin [3]. After is it produced, it can be cross-linked both chemically (with the use of glutaraldehyde) and with UV light in order to create hydrogels to serve as scaffolds for cell growth.

Figure 1.

The sequence for this part originated from the S. pyogenes native protein sequence [4], and was codon-optimized [2] for expression in E. coli.

References:

[1] An, B., Kaplan, D. L., & Brodsky, B. (2014). Engineered recombinant bacterial collagen as an alternative collagen-based biomaterial for tissue engineering. Frontiers in Chemistry, 2, 40. https://doi.org/10.3389/fchem.2014.00040

[2] Codon Optimization. (n.d.). VectorBuilder. Retrieved from https://en.vectorbuilder.com/tool/codon-optimization.html.

[3] Parenteau-Bareil, R., Gauvin, R., & Berthod, F. (2010). Collagen-Based Biomaterials for Tissue Engineering Applications. Materials, 3(3), 1863–1887. https://doi.org/10.3390/ma3031863

[4] UniProtKB - Q8RLX7. (n.d). UniProt. Retrieved from https://www.uniprot.org/uniprot/Q8RLX7.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 397
  Illegal PstI site found at 406
  Illegal PstI site found at 415
  Illegal PstI site found at 469
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 397
  Illegal PstI site found at 406
  Illegal PstI site found at 415
  Illegal PstI site found at 469
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 207
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 397
  Illegal PstI site found at 406
  Illegal PstI site found at 415
  Illegal PstI site found at 469
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 397
  Illegal PstI site found at 406
  Illegal PstI site found at 415
  Illegal PstI site found at 469
  Illegal NgoMIV site found at 517
  Illegal NgoMIV site found at 751
  Illegal NgoMIV site found at 1230
  Illegal AgeI site found at 1237
  
• 1000
  COMPATIBLE WITH RFC[1000]