Chiba/Goal

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[[Chiba|Introduction]] | [[Chiba/Project_Design|Project Design]] ( [[Chiba/Engeneering_Flagella|1.Sticky Hands]] | [[Chiba/Communication|2.Communication]] | [[Chiba/Quorum_Sensing|3.Size Control]] ) |  [[Chiba/Making Marimo|Making Marimos]] |  [[Chiba/Goal|Our Goal]] || [[Chiba/Team_Members|Team Members]] | [[Chiba/Members_Only|メンバ連絡簿]]
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==Our Goal==
As Final-Construction, we  will carry out four experiments.
*A test of adsorption between flagella
*A test to confirm a limit of size
*A test to form Marimo
*A test of size control



===A test of adsorption between flagella===
[[Image:Imida2.gif|frame]]
====Purpose====
Confirm adsorption between senders and receivers with His-Tag.

====Method====
#Culture senders and receivers in a test tube. 
#Drop senders into receivers and mix cobalt ion. <!-- They start to aggregate because of they have His-Tag. -->
#<!-- After aggregation, we -->Divide it into two groups. One is added imidazole and the other is non imidazole. Inoculate them on the plates.
#Check the colony. 
=====Prediction=====
If succeeded, cultures which is added imidazole will form a indipendent colony of senders&receivers. The other (non imidazole) will form colonies of adsorbed senders and receivers.

 1.Senderとreceiverをそれぞれ培養し、receiverの培養液の中に、senderを１滴加える。
 2.その中に金属イオンを加えると、senderとreceiverが鞭毛同士で吸着し始める。
 3.ある程度吸着したら、imidazolを加えたものとそうでないものをプレートにまく。
 4.imidazolを加えたものはばらばらのコロニーができ、imidazolなしのものは、senderとreceiverが
   くっついているので、くっついたコロニーができる。

===A test to confirm a limit of size===

====Purpose====
The purpose of this test is to confirm that Marimo has a limit of size on a plate.

====Method====


#Culture senders and receivers in a test tube. 
#Inoculate receivers equally on the plate, and drop senders at the center of it.
#After over night,pick colonies which are near the senders (colonies express GFP), the end of the circle expressed GFP, and out of the green circle.
#Dilute them.
#Mix cobalt ion into dilutions and inoculate on the plates.
#Check the colony.
=====Prediction=====
If succeeded,receivers which don’t express FliC (they didn’t sense AHL and express GFP) form colonies one by one. <br>And receivers expressed FliC (they sensed AHL and expressed GFP) form aggregated colonies.)

<br>[[Image:ugen.gif]]<br>



'''↑プレートにまく前にイミダゾールを入れる/入れないの条件の違いが書いてないよ！？by tashiro'''
'''↑イミダゾールを混ぜる実験はadsorption test です。'''

'''↑スペリングミス。inculateではなくincubateですby とよたろ'''

　

===A test to form Marimo===

[[Image:Howto.gif|frame]]
====Purpose====
Actually we make Marimos.
*AHL diffuse fast because it is very small molecule.
*Packed senders into a very thin capillary . AHL from it diffuse slowly and form concentrate gradients. Concentrate gradients enable Marimos to aggregate like a real ball.
(AHLはとても小さな分子で、自由度が高いので、拡散するのが早い。
キャピラリーを利用することによって、senderが生産するAHLの拡散を遅くし、AHLの濃度勾配を作ることができる。濃度勾配によってより球体に近いマリモができる。)

====Method====
#Culture senders and receivers in each test tube and mix Cobalt ions in receiver's tube.
#Add senders to receivers with a capillary.
#Check Marimos.
=====Prediction=====
If succeeded,AHL from senders spreads.<br>
Receivers express GFP and FliC-His,and besides,bond with cobalt ions.
<br>Finnally,form marimos.<br><br><br><br><br>

===A test of size control===
[[Image:size.gif|frame]]
====Purpose====
Control marimo's size by changing　the ratio of amount of senders and receivers.

====Method====
#Culture senders and receivers in each test tube. 
#Observe marimo's size by changing the amount of receivers and their types.(const.senders)
#Observe marimo's size by changing the amount of senders.(const.receivers)

=====Prediction=====
Low concentration of senders or  receivers : Mini Marimo<br>
High concentration of senders or receivers : Big Marimo

==Future==
*説明がどれも不十分なので、もしどこが面白いのか充分に書けないのならば、この箇所はカットしましょう。byとよたろ
*Play catch with a big marimo.(サイズをもっと大きくしてマリモでキャッチボールがしたい)
*Make marimo in liposome.(リポソームの中でマリモを作る)
*Biochromatography using gelled cells.(ゲル化細胞を利用した、バイオクロマトグラフィ） 



<!--ゲル化した細胞を利用した，バイオクロマトグラフフィ-->
<!--collect ion ? by tahiro-->
[[Image:biochromatography.jpg|frame]]