Part:BBa_K2220018
Display mCherry protein
It can display mCherrry 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.
mCherry is a monomeric fluorescent construct with peak excitation/emission at 587 nm/610 nm, respectively. It is resistant to photobleaching and is stable. It matures quickly, with a t0.5 of 15 minutes, allowing it to be visualised soon after translation.
This part can express AGA2-mCherry fusion protein, which can display mCherry onto the yeast cell wall.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 325
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 388
Illegal XhoI site found at 1102 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1171
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1153
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