Composite

Part:BBa_K5382160:Design

Designed by: yuyanyan chen   Group: iGEM24_HUBU-4-CHN   (2024-09-26)
Revision as of 12:36, 2 October 2024 by Chenyuyanyan (Talk | contribs) (→‎Source)


InaK-linker-4z 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 2299
  • 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
    Illegal SapI.rc site found at 1951


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-4z domain as well as the amount of final expression also need to be detected and controlled, including the subsequent purification steps.The experiment of this part is on going.

Source

The InaK is an ice nucleated protein from Pseudomonas syringae KCTC1832. The linker can be a natural amino acid sequence or an artificial design. A flexible linker is usually 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 containing approximately 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).
The 4z domain is a tetramer of the z domain, which is obtained by the z domain concatenated by GS linker and expressed in series by the coding sequence on the plasmid.

References