Development of universal baculovirus vector system for synthetic biology

• Project/Area Number: 18K14686
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 43060:System genome science-related
• Research Institution: Kumamoto University (2019-2020); Kyushu University (2018)
• Principal Investigator: TATSUKE Tsuneyuki 熊本大学, 大学院先端科学研究部(工), 特任准教授 (80637843)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: バキュロウイルス / ゲノム科学 / ウイルスベクター / バイオインフォマティクス / 生物物理学 / 最小ゲノム / 必須遺伝子 / 合成生物学

Outline of Final Research Achievements

This study aimed to reconstruct baculoviruses as versatile virus vectors for synthetic biology. I inserted several short DNA fragments in vitro into baculovirus genomic DNA to generate genetically minimum baculovirus genomic DNA. From them, I produced a polygene-deficient baculovirus library that can multiply in cultured insect cells. This process was repeated until viral replication was not observed, and a genetically minimized polygene-deficient baculovirus genomic DNA library was generated. This library is thought to contain several candidates for the prototype of next-generation general-purpose baculovirus vectors for synthetic biology.

Academic Significance and Societal Importance of the Research Achievements

昆虫を宿主とするバキュロウイルスは高等真核生物での遺伝子発現ベクターとしてバイオ医薬品の生産などで利用されている。本研究はバキュロウイルスを汎用遺伝子発現ベクターとして最大限に利用するために様々な多遺伝子破壊バキュロウイルスをライブラリとして作製した。この多遺伝子破壊バキュロウイルスライブラリは様々な用途に使用できる原型となる多遺伝子破壊バキュロウイルスを含んでいると考えられ、バキュロウイルスの多用途化に貢献できると期待される。

Research Products

• [Journal Article] Mild inactivation and inhibition of phenoloxidase in silkworm serum for xeno-free culture of insect cells (2020)
  • Author(s): Hino Masato、Karasaki Noriko、Tatsuke Tsuneyuki、Morokuma Daisuke、Masuda Akitsu、Mon Hiroaki、Lee Jae Man、Kusakabe Takahiro
  • Journal Title: Journal of insect biotechnology and sericology Volume: 89 Pages: 9-16
  • NAID: 130007821890
  • Peer Reviewed
• [Journal Article] Functional horseradish peroxidase?streptavidin chimeric proteins prepared using a silkworm-baculovirus expression system for diagnostic purposes (2019)
  • Author(s): Patmawati、Minamihata Kosuke、Tatsuke Tsuneyuki、Lee Jae Man、Kusakabe Takahiro、Kamiya Noriho
  • Journal Title: Journal of Biotechnology Volume: 297 Pages: 28-31
  • DOI: 10.1016/j.jbiotec.2019.03.007
  • Peer Reviewed
• [Journal Article] Expression and Purification of Vaccinia Virus DNA Topoisomerase IB Produced in the Silkworm-Baculovirus Expression System (2019)
  • Author(s): Xu Jian、Lee Jae Man、Tatsuke Tuneyuki、Ebihara Takeru、Masuda Akitsu、Hino Masato、Morokuma Daisuke、Fujita Ryosuke、Mon Hiroaki、Kusakabe Takahiro、Takahashi Masateru
  • Journal Title: Molecular Biotechnology Volume: 61 Issue: 8 Pages: 622-630
  • DOI: 10.1007/s12033-019-00184-4
  • Peer Reviewed
• [Presentation] 焼きなまし法による概日振動体常微分方程式モデルの拡張 (2019)
  • Author(s): 田附 常幸
  • Organizer: 第８回 九州山口沖縄リズム研究会