التفاصيل البيبلوغرافية
| العنوان: |
A Yeast Modular Cloning (MoClo) Toolkit Expansion for Optimization of Heterologous Protein Secretion and Surface Display in Saccharomyces cerevisiae |
| المؤلفون: |
Nicola M. O’Riordan, Vanja Jurić, Sarah K. O’Neill, Aoife P. Roche, Paul W. Young |
| سنة النشر: |
2024 |
| المجموعة: |
Smithsonian Institution: Figshare |
| مصطلحات موضوعية: |
Biochemistry, Medicine, Microbiology, Genetics, Molecular Biology, Biotechnology, Evolutionary Biology, Infectious Diseases, Biological Sciences not elsewhere classified, Chemical Sciences not elsewhere classified, yeast modular cloning, translational fusion partners, allow facile screening, achieving sufficient levels, secretion promoting signals, direct heterologous proteins, yeast surface display, yeast secretory pathway, widely used technique, moclo compatible parts, heterologous protein secretion, efficient surface display, cerevisiae expression cassettes, surface display, secretory pathway, surface expression, standardized parts, secretion efficiency, optimal secretion, efficient incorporation |
| الوصف: |
Saccharomyces cerevisiae is an attractive host for the expression of secreted proteins in a biotechnology context. Unfortunately, many heterologous proteins fail to enter, or efficiently progress through, the secretory pathway, resulting in poor yields. Similarly, yeast surface display has become a widely used technique in protein engineering but achieving sufficient levels of surface expression of recombinant proteins is often challenging. Signal peptides (SPs) and translational fusion partners (TFPs) can be used to direct heterologous proteins through the yeast secretory pathway, however, selection of the optimal secretion promoting sequence is largely a process of trial and error. The yeast modular cloning (MoClo) toolkit utilizes type IIS restriction enzymes to facilitate an efficient assembly of expression vectors from standardized parts. We have expanded this toolkit to enable the efficient incorporation of a panel of 16 well-characterized SPs and TFPs and five surface display anchor proteins into S. cerevisiae expression cassettes. The secretion promoting signals are validated by using five different proteins of interest. Comparison of intracellular and secreted protein levels reveals the optimal secretion promoting sequence for each individual protein. Large, protein of interest-specific variations in secretion efficiency are observed. SP sequences are also used with the five surface display anchors, and the combination of SP and anchor protein proves critical for efficient surface display. These observations highlight the value of the described panel of MoClo compatible parts to allow facile screening of SPs and TFPs and anchor proteins for optimal secretion and/or surface display of a given protein of interest in S. cerevisiae. |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
unknown |
| العلاقة: |
https://figshare.com/articles/journal_contribution/A_Yeast_Modular_Cloning_MoClo_Toolkit_Expansion_for_Optimization_of_Heterologous_Protein_Secretion_and_Surface_Display_in_Saccharomyces_cerevisiae/25408133Test |
| DOI: |
10.1021/acssynbio.3c00743.s001 |
| الإتاحة: |
https://doi.org/10.1021/acssynbio.3c00743.s001Test |
| حقوق: |
CC BY-NC 4.0 |
| رقم الانضمام: |
edsbas.B78EBCC8 |
| قاعدة البيانات: |
BASE |
