Coding

Part:BBa_K4323000

Designed by: Jessica Marinelli, Kevin Lim, Chun Hin Wong   Group: iGEM22_UManitoba   (2022-09-30)
Revision as of 19:43, 11 October 2022 by Nihnuhc (Talk | contribs)


Coding sequence for Aerolysin

Secreted, cytolytic toxin that forms pores in host membranes after proteolytic removal of a C-terminal propeptide, leading to destruction of the membrane permeability barrier and host cell death. The pores are formed by transmembrane beta-strands and are approximately 3 nm in diameter. This part has been codon optimized for expression in Escherichia coli.

Aerolysin toxicity

Figure 1. The toxicity of Aerolysin to Dreissena bugensis.

Ni-purified Aerolysin SDS-PAGE

Figure 2. His-Trap Ni-column purified Aerolysin. L: ladder, C5: 10 % B wash, D1: fraction of gradient at ~30 % B, D4: fraction of gradient at ~50 % B, D8: elution at 100 % B. A buffer: 20 mM Tris-HCl, 200 mM NaCl, pH 8.0; B buffer: 20 mM Tris-HCl, 200 mM NaCl, 300 mM imidazole, pH 8.0. Arrow: purified Aerolysin protein, MW: 55.2 kDa.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 206
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 75
    Illegal AgeI site found at 313
    Illegal AgeI site found at 1270
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None