Development of high-throughput human protein expression and purification system in silkworms

• Project/Area Number: 23760747
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biofunction/Bioprocess
• Research Institution: Shizuoka University
• Principal Investigator: TATSUYA Kato 静岡大学, (連合)農学研究科(研究院), 准教授 (00397366)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2012: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: カイコ / ハイスループット / タンパク質発現 / かいこさなぎ / FLAGタグ / タンパク質精製

Research Abstract

Multi-expression and purification of recombinant proteins in BmNPV bacmid-silkworm expression system were constructed. Many kinds of recombinant BmNPV bacmid DNA were prepared in a 96 well plate and recombinant proteins were also purified in a 96 well plate packing a purification gel. The further improvement of protein extraction and purification step leads to the establishment of a more practical silkworm system.

Research Products

• [Journal Article] Construction of new ligation-independent cloning vectors for the expression and purification of recombinant proteins in silkworms using BmNPV bacmid system (2013)
  • Author(s): Kato T, Thompson JR, Park EY
  • Journal Title: PLoS One Volume: Vol.8, No.5
  • NAID: 120005257829
  • Peer Reviewed
• [Journal Article] Construction of new ligation-independent cloning vectors for the expression and purification of recombinant proteins in silkworms using BmNPV bacmid system (2013)
  • Author(s): Tatsuya Kato, James R. Thompson, Enoch Y. Park
  • Journal Title: PLoS ONE Volume: 8 Issue: 5 Pages: e64007-e64007
  • DOI: 10.1371/journal.pone.0064007
  • Peer Reviewed
• [Presentation] マルチウェルプレートを用いたカイコ蛹での組換えタンパク質生産 (2013)
  • Author(s): 加藤竜也、トンプソンジェームス、朴龍洙
  • Organizer: 2013年度日本農芸化学会大会
  • Place of Presentation: 仙台市、東北大学
• [Presentation] Vector development for high-throughput protein expression in silkworms (2012)
  • Author(s): T. Kato
  • Organizer: IBS2012 15^<th> International Biotechnology Symposium and Exhibition
  • Place of Presentation: Daegu, South Korea
• [Presentation] Ligation-independent cloning法を用いたカイコ-バクミド発現用ベクターの構築 (2012)
  • Author(s): 加藤竜也、トンプソンジェームス、朴龍洙
  • Organizer: 2012年度日本農芸化学会大会
  • Place of Presentation: 京都市、京都女子大学
• [Presentation] マルチウェルプレートを用いたカイコ蛹での組換えタンパク質生産
  • Author(s): 加藤竜也、トンプソン ジェームス、朴 龍洙
  • Organizer: 日本農芸化学会2013年度大会
  • Place of Presentation: 東北大学川内北キャンパス（仙台）
• [Presentation] Ligation-independent cloning法を用いたカイコ-バクミド発現用ベクターの構築
  • Author(s): 加藤 竜也、トンプソン ジェームス、朴 龍洙
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大学（京都）