Alteration of Secretion Rates in Model Yeasts and Development of Assays for Genome-Wide Screening of Secretion

Дата публикации: 20-07-2026 00:00:00

In this work, I investigated strategies to improve recombinant protein production in yeast by targeting both transcriptional regulation and protein secretion. First, I characterized how mutations within the promoter region of Saccharomyces cerevisiae influence gene expression. Targeted mutagenesis revealed that increasing GC content within the TATA box and pre-initiation complex consistently reduced expression, identifying these regions as highly sensitive to sequence composition. I then expanded this approach using CRISPR-enabled trackable genome engineering (CREATE) to introduce a broader range of mutation types across an extended promoter region. This analysis confirmed that GC-rich substitutions were strongly associated with decreased expression, while certain AT- or mixed-base substitutions in the scanning region and transcriptional start site contributed to increased promoter activity. Finally, I adapted a mid-western (colony blot) secretion assay for use in Kluyveromyces...

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