Coding

Part:BBa_K2220005

Designed by: Nan Rong   Group: iGEM17_BNU-China   (2017-10-18)


Display PETase protein

It can display PETase on the surface of S.cerevisiae.

Among various GPI-anchored proteins, α/a-agglutinins are one of the most common proteins have been utilized for the display of proteins on a yeast cell surface. The native a and α agglutinin receptors are believed to act as adhesion molecules to stabilize cell-cell interactions during mating and to facilitate fusion between the a and α haploid yeast cells. A-agglutinin has a core subunit encoded by AGA1 and a binding subunit encoded by AGA2. Both a-agglutinin and the core subunit of a-agglutinin consist of a secretion-signal region, a functional region, a support region, and a putative GPI anchor-attachment signal. After genetic engineering, foreign protein can be fused to the C-terminal of the binding subunit of a-agglutinin. As AGA1 protein is secreted from the cell and becomes covalently attached to the β-glucan in the extracellular matrix of the yeast cell wall, AGA2 protein fused with foreign protein on its C-terminus will then bind to aga1p through two disulfides bonds. As a result, the heterologous proteins are covalently linked with glucan-layer.

Polyethyleneterephthalate (PET) is the condensation of terephthalic acid and ethylene glycol.PETase was found from a kind of microorganism living on PET as the main carbon source. It can degrade macromolecular polymers into monomers.

This part can express AGA2-PETase fusion protein, which can display PETase onto the yeast cell wall.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 325
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 472
    Illegal BamHI site found at 388
    Illegal XhoI site found at 1186
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1255
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1237


[edit]
Categories
Parameters
None