Elucidation of the molecular mechanism for highly active terminator regions in Saccharomyces cerevisiae

2014 Fiscal Year Final Research Report

• Project/Area Number: 25660071
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied microbiology
• Research Institution: Toyota Central R&D Lab., Inc.
• Principal Investigator: MATSUYAMA Takashi 株式会社豊田中央研究所, 先端研究センター バイオ研究室, 主任研究員 (90394882)
• Co-Investigator(Kenkyū-buntansha): KITAGAWA Takao 山口大学, 医学研究科, 助教 (20614928)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: 遺伝子発現制御 / ターミネーター / 出芽酵母

Outline of Final Research Achievements

In Saccharomyces cerevisiae, the activity of the top 5-ranked terminator regions to increase the protein accumulation of the upstream ORF conferred about twice the relative activity as the standard PGK1 terminator under two promoters and three upstream ORFs, under various mediums with a different carbon source, under various growth conditions, and in several yeast strains. Among the terminators in S. cerevisiae, the DIT1 terminator (DIT1t) had the highest activity and was the most versatile for metabolic engineering.
In addition, we discovered the molecular mechanism to increase the protein accumulation of the ORF upstream the DIT1t region, and found that NAB6 and PAP1 were genetically involved in this process via the specific cis element within the DIT1t region.

Free Research Field

応用微生物学