Difference between revisions of "Part:BBa K5382110:Design"

 
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===Design Notes===
 
===Design Notes===
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The two amino acid substitution of the z domain relative to the B domain is key, which may affect its binding affinity and specificity to IgG. The plasmid construction and introduction of Inak-linker-z domain as well as the amount of final expression also need to be detected and controlled, including the subsequent purification steps.
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The two amino acid substitution of the z domain relative to the B domain is key, which may affect its binding affinity and specificity to IgG. The plasmid construction and introduction of Inak-linker-z domain as well as the amount of final expression also need to be detected and controlled, including the subsequent purification steps.The experimental design of this part is another direction of our project, the plasmid has been successfully constructed, and the specific verification is ongoing
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Revision as of 15:46, 27 September 2024


Inak-linker-z domain_protein domain that have the ability to bind to lgG


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1674
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 722
    Illegal NgoMIV site found at 962
    Illegal NgoMIV site found at 1130
    Illegal AgeI site found at 517
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 692
    Illegal BsaI.rc site found at 1182
    Illegal SapI.rc site found at 326


Design Notes

The two amino acid substitution of the z domain relative to the B domain is key, which may affect its binding affinity and specificity to IgG. The plasmid construction and introduction of Inak-linker-z domain as well as the amount of final expression also need to be detected and controlled, including the subsequent purification steps.The experimental design of this part is another direction of our project, the plasmid has been successfully constructed, and the specific verification is ongoing


Source

Inak is a protein derived from Pseudomonas syringae KCTC1832, an ice nucleated protein Linker is composed of Gly and Ser. The z domain is derived from the B domain of protein A. Protein A is a protein widely found on the cell wall of Staphylococcus aureus. The extracellular portion of protein A contains five highly homologous IgG binding domains, named E, D, A, B, and C from the N-terminal, each domain contains about 58 amino acid residues. The z domain is an engineered analogue of the B domain, originally developed as an affinity purification treatment for fusion protein production. In contrast to the B domain, the z domain contains two amino acid substitutes (Ala1→Val and Gly29→Ala).

References