Coding

Part:BBa_K2220008

Designed by: Nan Rong   Group: iGEM17_BNU-China   (2017-10-19)


Display CBH protein

It can display xo-1,4-β-D-glucannase 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.

CBH gene can express xo-1,4-β-D-glucannase.

This part can express AGA2-CBH fusion protein, which can display xo-1,4-β-D-glucannase onto the yeast cell wall.

It can display exo-1,4-β-D-glucannase on the surface of S.cerevisiae.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 325
    Illegal NheI site found at 1239
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 388
    Illegal BamHI site found at 1113
    Illegal XhoI site found at 1459
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1528
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1510


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Categories
Parameters
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