Plant virus replication-mimicking expression vector for Saccharomyces Cerevisiae

2019 Fiscal Year Final Research Report

• Project/Area Number: 16K14909
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: Nishimiya Yoshiyuki 国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (00357716)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Keywords: 発現系 / 酵母

Outline of Final Research Achievements

Tobacco mosaic virus (TMV) is a positive-strand plant RNA virus, and it is known that TMV is able to replicate in cytoplasm of Saccharomyces Cerevisiae. Here, I tried to construct TMV replication-mimicking expression vector for S. Cerevisiae, which contains 5’-leader sequence (omega leader), RNA-dependent RNA polymerase (RdRp)-encoding gene, subgenomic promoters (sgps) for movement and coat proteins, and 3’UTR of TMV. For target proteins, cloning sites (css) were introduced into downstream of the sgps. Interestingly, when encoding gene for target protein is under the control of cp sgp, the protein was expressed without expression of RdRp. Further analysis of this phenomenon might be helpful in understanding replication system of RNA viruses.

Free Research Field

蛋白質工学

Academic Significance and Societal Importance of the Research Achievements

ゲノムRNAやサブゲノムRNAの複製を担うTMV由来のRdRpが発現されなくともcp sgp下にコードされたタンパク質が発現されるという予想外の結果がもたらされた。本来の宿主であるタバコのプロトプラストを使った研究では，RdRpの発現なしにはサブゲノムRNAが作られないことが報告されており，今回得られた結果が何に起因するかを解析することでウイルスが宿主の翻訳装置を乗っ取る仕組みに新たな知見をもたらすと期待される。