Team:Chiba/jk

From 2008.igem.org

(Difference between revisions)
(still under discussion)
(To Do List)
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===still under discussion===
===still under discussion===
*出力を交換できる形でクローニングしよう-->intermideatesの作成
*出力を交換できる形でクローニングしよう-->intermideatesの作成
-
*AHL synthase のkineticsについての情報は?
+
*AHL synthase のkineticsについての情報は?-->[[Team:Chiba/Reference/j|ref.]]
*ベクターの乗せ換え(PSB1-->P15A)(井山、小林、川崎)<Br>
*ベクターの乗せ換え(PSB1-->P15A)(井山、小林、川崎)<Br>
----
----

Revision as of 02:38, 18 September 2008

>Notebook


August
MTWTFSS
        [http://2008.igem.org/Team:Chiba/Calendar-Main/1_August_2008 1] [http://2008.igem.org/Team:Chiba/Calendar-Main/2_August_2008 2] [http://2008.igem.org/Team:Chiba/Calendar-Main/3_August_2008 3]
[http://2008.igem.org/Team:Chiba/Calendar-Main/4_August_2008 4] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/5_August_2008&action=edit 5] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/6_August_2008&action=edit 6] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/7_August_2008&action=edit 7] [http://2008.igem.org/Team:Chiba/Calendar-Main/8_August_2008 8] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/9_August_2008&action=edit 9] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/10_August_2008&action=edit 10]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/11_August_2008&action=edit 11] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/12_August_2008&action=edit 12] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/13_August_2008&action=edit 13] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/14_August_2008&action=edit 14] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/15_August_2008&action=edit 15] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/16_August_2008&action=edit 16] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/17_August_2008&action=edit 17]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/18_August_2008&action=edit 18] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/19_August_2008&action=edit 19] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/20_August_2008&action=edit 20] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/21_August_2008&action=edit 21] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/22_August_2008&action=edit 22] [http://2008.igem.org/Team:Chiba/Calendar-Main/23_August_2008 23] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/24_August_2008&action=edit 24]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/25_August_2008&action=edit 25] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/26_August_2008&action=edit 26] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/27_August_2008&action=edit 27] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/28_August_2008&action=edit 28] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/29_August_2008&action=edit 29] [http://2008.igem.org/Team:Chiba/Calendar-Main/30_August_2008 30] [http://2008.igem.org/Team:Chiba/Calendar-Main/31_August_2008 31]
September
MTWTFSS
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/1_September_2008&action=edit 1] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/2_September_2008&action=edit 2] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/3_September_2008&action=edit 3] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/4_September_2008&action=edit 4] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/5_September_2008&action=edit 5] [http://2008.igem.org/Team:Chiba/Calendar-Main/6_September_2008 6] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/7_September_2008&action=edit 7]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/8_September_2008&action=edit 8] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/9_September_2008&action=edit 9] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/10_September_2008&action=edit 10] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/11_September_2008&action=edit 11] [http://2008.igem.org/Team:Chiba/Calendar-Main/12_September_2008 12] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/13_September_2008&action=edit 13] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/14_September_2008&action=edit 14]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/15_September_2008&action=edit 15] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/16_September_2008&action=edit 16] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/17_September_2008&action=edit 17] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/18_September_2008&action=edit 18] [http://2008.igem.org/Team:Chiba/Calendar-Main/19_September_2008 19] [http://2008.igem.org/Team:Chiba/Calendar-Main/20_September_2008 20] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/21_September_2008&action=edit 21]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/22_September_2008&action=edit 22] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/23_September_2008&action=edit 23] [http://2008.igem.org/Team:Chiba/Calendar-Main/24_September_2008 24] [http://2008.igem.org/Team:Chiba/Calendar-Main/25_September_2008 25] [http://2008.igem.org/Team:Chiba/Calendar-Main/26_September_2008 26] [http://2008.igem.org/Team:Chiba/Calendar-Main/27_September_2008 27] [http://2008.igem.org/Team:Chiba/Calendar-Main/28_September_2008 28]
[http://2008.igem.org/Team:Chiba/Calendar-Main/29_September_2008 29] [http://2008.igem.org/Team:Chiba/Calendar-Main/30_September_2008 30]
October
MTWTFSS
    [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/1_October_2008&action=edit 1] [http://2008.igem.org/Team:Chiba/Calendar-Main/2_October_2008 2] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/3_October_2008&action=edit 3] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/4_October_2008&action=edit 4] [http://2008.igem.org/Team:Chiba/Calendar-Main/5_October_2008 5]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/6_October_2008&action=edit 6] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/7_October_2008&action=edit 7] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/8_October_2008&action=edit 8] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/9_October_2008&action=edit 9] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/10_October_2008&action=edit 10] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/11_October_2008&action=edit 11] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/12_October_2008&action=edit 12]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/13_October_2008&action=edit 13] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/14_October_2008&action=edit 14] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/15_October_2008&action=edit 15] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/16_October_2008&action=edit 16] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/17_October_2008&action=edit 17] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/18_October_2008&action=edit 18] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/19_October_2008&action=edit 19]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/20_October_2008&action=edit 20] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/21_October_2008&action=edit 21] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/22_October_2008&action=edit 22] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/23_October_2008&action=edit 23] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/24_October_2008&action=edit 24] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/25_October_2008&action=edit 25] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/26_October_2008&action=edit 26]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/27_October_2008&action=edit 27] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/28_October_2008&action=edit 28] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/29_October_2008&action=edit 29] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/30_October_2008&action=edit 30] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/31_October_2008&action=edit 31]
November
MTWTFSS
          [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/1_November_2008&action=edit 1] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/2_November_2008&action=edit 2]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/3_November_2008&action=edit 3] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/4_November_2008&action=edit 4] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/5_November_2008&action=edit 5] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/6_November_2008&action=edit 6] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/7_November_2008&action=edit 7] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/8_November_2008&action=edit 8] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/9_November_2008&action=edit 9]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/10_November_2008&action=edit 10] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/11_November_2008&action=edit 11] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/12_November_2008&action=edit 12] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/13_November_2008&action=edit 13] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/14_November_2008&action=edit 14] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/15_November_2008&action=edit 15] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/16_November_2008&action=edit 16]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/17_November_2008&action=edit 17] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/18_November_2008&action=edit 18] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/19_November_2008&action=edit 19] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/20_November_2008&action=edit 20] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/21_November_2008&action=edit 21] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/22_November_2008&action=edit 22] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/23_November_2008&action=edit 23]
[http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/24_November_2008&action=edit 24] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/25_November_2008&action=edit 25] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/26_November_2008&action=edit 26] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/27_November_2008&action=edit 27] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/28_November_2008&action=edit 28] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/29_November_2008&action=edit 29] [http://2008.igem.org/wiki/index.php?title=Team:Chiba/Calendar-Main/30_November_2008&action=edit 30]

Contents

To Do List

  • 出力の決定(杉山、福冨)
    • LacZ-->基質濃度に依存
    • rLuciferine-->見えない?-->基質が必要、光分解性(詳しく調べる)
  • Rs,PFL(LuxR,J3720-->imperial,2006)の作成(杉山、福冨)
  • 入力の決定(香取)
    • Arabinose promotor?-->線形成?On/Offはっきり?
    • On/Offの必要性は-->なし?
  • Time-delayの確認(井山、川崎、小林、冨永)
    • cinIのクローニング(井山、川崎)
    • 差を広げる-->温度依存性、培地依存性.PFL(Positive feedback loop)(BBa_J37015,BBa_J37020)の利用.
    • AHL固定(1nM,10nM,100nM~)-->閾値(10nM付近以上)を超えると、GFPによる蛍光強度は変わらない。4~5時間で飽和する。
    • 固体培地にspot
    • His TagつきGFP(その他タンパク質)を発現-->freeze-->SDS-PAGE-->blotting
    • Is(BW,XL10G)をT9002(BW)でdelay check
      • さらに-BBa_J37020を形質転換(XL10G,BW)でtime-delayをチェック
  • work out deal to print T-shirts (井山、福冨)

still under discussion

  • 出力を交換できる形でクローニングしよう-->intermideatesの作成
  • AHL synthase のkineticsについての情報は?-->ref.
  • ベクターの乗せ換え(PSB1-->P15A)(井山、小林、川崎)

実験について(どこにおいたらいいか分からなかったので~不必要かも)

時間スイッチ作成

発現時間の調節をするために、以下の要件が挙げられる。

  1. インデューサーの合成速度を下げる。
    RBSの調整(B0030-->B0033),コピー数の調整(5-->300)
  2. レセプタータンパク質の感度を下げる。
    クオラムセンシングのクロストークを利用する。生物によってAHL(Acyl-L-Homoserine Lactone)のアシル基が異なる(ref.3).生物ごとのクオラムセンシングについては、ref.9参照.
  • ex)( )内はシンターゼ
    • P.aeruginosa->C4-AHL(RhlI)
    • V. fisheri->C6-3-oxo-AHL(LuxI)
    • A.tumefaciens->C8-3-oxo-AHL(TraI)
    • P.aeruginosa->C10-3-oxo-AHL(VanI)
    • P.aeruginosa->C12-3-oxo-AHL(LasI)
    • R.leguminosarum->C14-3-hydroxy-AHL(CinI)

Construction(From BioBrick)

  • Sender(Autoinducer synthase)
LuxI,LasI,CinI,RhlI(lacプロモーター)
  • Receiver
LuxR,LasR,CinR,RhlR

Cross-Talk Check

  1. LuxR vs LuxI(OK),LasI(OK),CinI(?),RhlI(OK)
  2. LuxI vs LuxR,LasR,CinR,RhlR

RBS交換

  • 既存のパーツ
    • B0034+luxI
  • New parts(planning)
    • B0033(weak)+luxI
    • B0032(medium)+luxI
    • B0031(weaker)+luxI


連絡

実験結果より -->Rs濃度が律速→PFL(J37020)使う?Rs替える?




入力班 送受信班 出力班 その他

小林あおい Aoi Kobayashi

香取崇広 Takahiro Katori

冨永将大 Masahiro Tominaga

川崎浩平 Kohei Kawasaki

井山佳美 Yoshimi Iyama

福冨浩樹 Hiroki Fukutomi

杉山まい Mai Sugiyama


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