Difference between revisions of "Part:BBa K4429003:Design"

 
Line 9: Line 9:
 
Ensuring the correct V->C region interface. Used Uniprot entry for human IGHG1 to find where constant region starts.
 
Ensuring the correct V->C region interface. Used Uniprot entry for human IGHG1 to find where constant region starts.
  
 +
Removing BioBrick illegal restriction sites using silent mutation in SnapGene.
 +
 +
Mutation of Val214 to a GTT [2]
  
  
 
===Source===
 
===Source===
  
−
Variable region protein sequence from PDB, reverse translated using E. coli codon usage table in SnapGene, constant region from GB: MT470197.1. The entire heavy chain was then codon optimised for E. coli K12 using IDT codon optimisation tool.
+
Variable region protein sequence from PDB 7CTH [1], reverse translated using E. coli codon usage table in SnapGene, constant region from GenBank, accession no: MT470197.1. The entire heavy chain was then codon optimised for E. coli K12 using IDT codon optimisation tool.
  
 
===References===
 
===References===
 +
[1] https://www.rcsb.org/structure/7CTH
 +
 +
[2] Leith, E. M., O'Dell, W. B., Ke, N., McClung, C., Berkmen, M., Bergonzo, C., Brinson, R. G., & Kelman, Z. (2019). Characterization of the internal translation initiation region in monoclonal antibodies expressed in Escherichia coli. The Journal of biological chemistry, 294(48), 18046–18056. https://doi.org/10.1074/jbc.RA119.011008

Revision as of 02:52, 14 October 2022


C10 IgG1 HC


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Ensuring the correct V->C region interface. Used Uniprot entry for human IGHG1 to find where constant region starts.

Removing BioBrick illegal restriction sites using silent mutation in SnapGene.

Mutation of Val214 to a GTT [2]


Source

Variable region protein sequence from PDB 7CTH [1], reverse translated using E. coli codon usage table in SnapGene, constant region from GenBank, accession no: MT470197.1. The entire heavy chain was then codon optimised for E. coli K12 using IDT codon optimisation tool.

References

[1] https://www.rcsb.org/structure/7CTH

[2] Leith, E. M., O'Dell, W. B., Ke, N., McClung, C., Berkmen, M., Bergonzo, C., Brinson, R. G., & Kelman, Z. (2019). Characterization of the internal translation initiation region in monoclonal antibodies expressed in Escherichia coli. The Journal of biological chemistry, 294(48), 18046–18056. https://doi.org/10.1074/jbc.RA119.011008