Part:BBa_K4430007:Design
GFP-CBD
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 829
Illegal AgeI site found at 811 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 641
Design Notes
In general, endolysin-targeting Gram-positive bacteria are composed of N-terminal enzymatic activity domains (EADs) with bacterial cell-lysing capability and C-terminal cell-binding domain (CBD) for adhering to the bacterial cell wall. Because the total amount of CBD binding sites on a single cell can be 107 or more, the binding capability of CBD is comparable to that of an antibody. Researchers had shown that CBD of bacteriophage P108 binds strongly to the cell surfaces of SA. GFP emits green fluorescence under the confocal laser scanning microscope. Using fusion protein GFP-CBD, we can confirm whether CBD binds strongly to the cell surfaces of SA. 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 are proven helpful for stabilizing such types of neighbors by facilitating the covalent bonding among them. So the flexible linker GGSG was used as fusion protein linker in GFP-CBD.
Source
GFP is from BBa_K404316. Protein linker is from BBa_K4430004. CBD is from BBa_K4430002.