Part:BBa_K5143010

Alpha-Factor secretion signal V2 optimized for expression in Saccharomyces cerevisiae

Description

The α-mating factor is a peptide hormone produced by the yeast Saccharomyces cerevisiae, which is crucial for the mating process of these yeast cells. Here are the key details about the α-mating factor:

• Function: It is involved in the mating process of S. cerevisiae by being secreted by α-type yeast cells to attract a-type cells, facilitating the mating interaction.
• Production: Synthesized as a pre-protein that includes a signal peptide and the mature α-mating factor peptide.
• Structure and Processing:
  • Pre-Pro-α-Mating Factor: Initially synthesized with a signal peptide (pre-region) that directs the nascent protein to the secretory pathway.
  • Mature Peptide: After cleavage of the signal peptide, the remaining peptide is processed into the active α-mating factor.
  • Processing: Includes cleavage of the signal peptide and pro-region to yield the mature peptide.
• Secretion:
  • Signal Peptide Function: Directs the protein to the endoplasmic reticulum (ER), where it is cleaved for passage through the secretory pathway.
  • Mature Peptide Secretion: Transported through the Golgi apparatus and secreted into the extracellular environment.
• Applications in Biotechnology:
  • Protein Secretion: The α-mating factor's signal peptide is used to enhance the secretion of various proteins in yeast by attaching it to a protein of interest.
  • Engineering Yeast Strains: Yeast strains can be engineered for efficient production of heterologous proteins by incorporating the α-mating factor signal peptide into their expression constructs.

Construction

The Alpha-Factor gene was synthesised and its nucleotide sequence optimised for DNA synthesis and expression in Saccharomyces cerevisiae. The gene exists in 2 versions: which limits homologous recombination in the Saccharomyces cerevisiae genome. The second version can be found here : BBa_K5143009 These two versions of the alpha-factor can be found in the following composite part:

References

Brake, A. J. et al. Alpha-factor-directed synthesis and secretion of mature foreign proteins in Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences 81, 4642–4646 (1984). Aza, P., Molpeceres, G., de Salas, F. & Camarero, S. Design of an improved universal signal peptide based on the α-factor mating secretion signal for enzyme production in yeast. Cell Mol Life Sci 78, 3691–3707 (2021).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 51
  Illegal NheI site found at 211
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]