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Chiba iGEM 2007

Chiba iGEM 2007. Marimo Bacteria. Concept. Marimo Chiba. 毬藻 -. Same Background: Applied Chemistry. Daisuke Umeno Taro Toyota. Hiroki Fukutomi Marina Gushiken Risa Kajiwara. Mutsuko Kazama Yukihiro Takami Masahiro Tominaga. Concept. Marimo?. Marimo is… a green spherical algae

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Chiba iGEM 2007

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  1. Chiba iGEM 2007 Marimo Bacteria Chiba University, Japan

  2. Concept Marimo Chiba 毬藻- Same Background: Applied Chemistry • Daisuke Umeno • Taro Toyota • Hiroki Fukutomi • Marina Gushiken • Risa Kajiwara • Mutsuko Kazama • Yukihiro Takami • Masahiro Tominaga Chiba University, Japan

  3. Concept Marimo? • Marimo is… • a green spherical algae • popular in Japan as a National Tresure • marvelous velvet • Motivations -Why marimo? • Spherical Multicellular Organism!? Chiba University, Japan

  4. Marimo? Concept • How the natural marimos are made Core Glow 絵は要変更 Chiba University, Japan

  5. Design Concept 1.Make the core by sticking sticked Core Chiba University, Japan

  6. Design Concept 2.Insert the sender core into the bacteria culture. Core Culture Chiba University, Japan

  7. Culture Design Concept 4. sticked Core Chiba University, Japan

  8. Culture Design Concept 4+a. The size of marimos could be controled by tuning the signal gradient. Complete! Core Chiba University, Japan

  9. Communication Module Sticky Power Boundary Maker Switching threshold [AHL] Affinity Tag Quorum Sensing Tuning Quorum Sensing Requirements Concept Communication Module Sticky Power Boundary Maker • FliC-Hisの作成 • 発現の確認 • 吸着の確認 Marimo Chiba University, Japan

  10. Switching threshold [AHL] Affinity Tag Quorum Sensing Tuning Quorum Sensing Requirements Concept Communication Module   Sticky Power Boundary Maker    (parallelism?) Marimo Chiba University, Japan

  11. Communication Module Sticky Power Sticky Power Size Control Switching threshold [AHL] Affinity Tag Quorum Sensing Tuning Quorum Sensing Requirements Concept Communication Module Size Control • FliC-Hisの作成 • 発現の確認 • 吸着の確認 Marimo Chiba University, Japan

  12. histidine Co2+ Affinity Tag 1.Display 2.Affinity His-tag with Co2+ Affinity Tag What needed to stick bacteria: Display & Affinity Co2+ On flagellar fillaments Chiba University, Japan

  13. Affinity Tag Display & Affinity 1.Display: Peptide insertable D3 可変domain D3 D1 D2 His-Tag (1-10 flagella per cell) D0 http://www.npn.jst.go.jp/index_jp.html Chiba University, Japan

  14. Affinity Tag Affinity Check Method: Beads Adsorption Beads 脱落 Chiba University, Japan

  15. -GI826(fliC,motB) Affinity Tag Affinity Check Method: Beads adsorption Exp. Affinity check Strain -JW1908(fliC) Plasmids -FliC-His -No plasmid * motB deletion provides cell with the flagella completely assembled but not rotating. Worked Out! Chiba University, Japan

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