Composite

Part:BBa_K4430005:Design

Designed by: Ruyi Shi   Group: iGEM22_GYHS   (2022-09-20)


GFP-gp13


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1211
    Illegal BglII site found at 2885
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2063
    Illegal AgeI site found at 1334
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 641
    Illegal BsaI.rc site found at 2259


Design Notes

Gp13 is a receptor binding protein (RBP) from the genome of E. coli O157 phage EP335. GFP emits green fluorescence under the confocal laser scanning microscope. Using fusion protein GFP-gp13, we can confirm whether gp13 binds strongly to the cell surfaces of E. coli O157 strains. Commonly, when interdomain movement and association of fusion protein is desired, flexible linkers are a good choice. Flexible linkers are usually passive; they serve by retaining distance between functional protein motifs. The length of flexible linkers is the key property that determines the appropriate folding in fusion proteins, if not achieved, leads to the formation of non-functional aggregates. The combination of non-polar Gly residues with the polar Ser or Thr is routinely used for synthesizing the flexible linkers. The flexible linkers is proven helpful for stabilizing such types of neighbors by facilitating the covalent bonding among them. So the flexible linker GGSG was used as fusion proteins linker in GFP-gp13.


Source

GFP is from BBa_K404316. Protein linker is from BBa_K4430004. Gp13 is from BBa_K4430000.

References