Selection of terminator that drives high-level expression of introduced genes in the model diatom Phaeodactylum tricornutum

2022 Fiscal Year Final Research Report

• Project/Area Number: 21K19149
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 40:Forestry and forest products science, applied aquatic science, and related fields
• Research Institution: Kochi University
• Principal Investigator: Adachi Masao 高知大学, 教育研究部自然科学系農学部門, 教授 (70274363)
• Project Period (FY): 2021-07-09 – 2023-03-31
• Keywords: 珪藻 / ターミネーター / 形質転換

Outline of Final Research Achievements

Promoters and terminators are key elements for the regulation of gene expression. Recently, we investigated the activity of promoters derived from marine diatom-infecting viruses (DIVs) in marine diatoms. Then, we obtained DIV promoter, CsetP4 having high activity. However, the terminators that can express the transgene at high levels for use in combination with CsetP4 have not been investigated. In this study, we investigated the activities of three terminators, two endogenous terminators from diatoms (PtfcpA terminator derived from Phaeodactylum tricornutum and CffcpA terminator derived from Cylindrotheca fusiformis) and a viral terminator, CsetT4 derived from DIV used together with CsetP4. The change in the expression level of the transgene by the terminator used was observed. Among three combinations, the combination of CsetP4 and PtfcpA terminator leads to an increase in the expression level of the transgene.

Free Research Field

海洋微生物学

Academic Significance and Societal Importance of the Research Achievements

本研究の成果は、ターミネーターが導入遺伝子の発現に及ぼす影響の解明の一助となったと考えられる。また、構築した導入遺伝子の高発現システムを用いることにより、バイオ燃料やワクチンや抗体などの有用物質の大量生産が期待される。さらに、高発現システムを国内外で知的財産化することにより、バイオ産業の振興や国際競争力の向上が期待される。また、珪藻によるこれらの有用物質生産の際には、光合成による炭酸ガス固定を伴うことから、地球温暖化防止効果が期待され、国連が掲げるSDGsのうち、「7.エネルギーをクリーンに」と「13.気候変動に具体的な対策を講じること」の2つのチャレンジに貢献するものと考えられる。