Exploration and identification of functional genes from subseafloor microorganisms based on substrate response phenomena

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K06828
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 45040:Ecology and environment-related
• Research Institution: Kochi University
• Principal Investigator: WAKAMATSU Taisuke 高知大学, 教育研究部総合科学系生命環境医学部門, 准教授 (60597938)
• Co-Investigator(Kenkyū-buntansha): 諸野 祐樹 国立研究開発法人海洋研究開発機構, 超先鋭研究開発部門(高知コア研究所), 主任研究員 (30421845)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 遺伝子 / DNA / 海底下微生物 / 基質応答 / ハロゲン化合物 / 金属イオン

Outline of Final Research Achievements

First, two metagenomic SIGEX libraries were constructed using evoglow gene as a reporter gene for subseafloor sediments. Next, SIGEX screening using five aromatic compounds (benzoate, 3-chlorobenzoate, 3-hydroxybenzoate, phenol, and 2,4-dichlorophenol) under aerobic and anaerobic conditions revealed significant differences in the inducible clone ratios under these conditions. 3-Chlorobenzoate and 2,4-dichlorophenol led to a higher induction ratio than that for the other non-chlorinated aromatic compounds under both aerobic and anaerobic conditions. After the further screening of induced clones, four clones induced by 3-chlorobenzoate only under anaerobic conditions were isolated and characterized. Furthermore, 408 clones were isolated that may respond to 2,4-dichlorophenol, MnCl2, or NiCl2 under aerobic conditions.

Free Research Field

生化学

Academic Significance and Societal Importance of the Research Achievements

evoglowをレポーター遺伝子とすることで好気/嫌気両条件下でSIGEX応答解析が可能になった。得られた基質応答DNA断片はほぼ(機能)未知遺伝子群のみから構成されており、バイオインフォマティクスなど他の方法では取得不可能であった。本研究成果により海底下生命圏のみならず他の生命圏にも存在する機能未知遺伝子のSIGEX法を用いた探索や同定の普及に大きく繋がると考える。