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<h3><p>&nbsp; DNAの相補的塩基対に着目した。その二本の一本鎖DNAはある程度塩基対がミスマッチしていても結合することができる。そのミスマッチの塩基対が少ないほど結合力が高い。この優劣を用いて黒髭危機一髪の作製を試みた。<br>
<h3><p>&nbsp;We paid attention to complementary base pair of DNA Single strand DNA of two can be connected even if I make a mistake, and base pair matches it to some extent. The fewer and fewer mismatched base pair, Cohesion is high. We tried to construct the Pop-up Pirate using DNA and porous silica particle.  <br>


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<td><div style="width: 330px;"><img alt="" src="/images/b/b7/Fitfiani.gif" width="320" height="240" border="0" / ><p>(*^_^*)</p><div></td></table>
<td><div style="width: 330px;"><img alt="" src="/images/b/b7/Fitfiani.gif" width="320" height="240" border="0" / ><p>(*^_^*)</p><div></td></table>


&nbsp; 黒髭危機一髪とは日本発祥の玩具である。土台の樽と飛び出す黒髭人形から成り立ち、土台に剣を刺し、特定の箇所に刺すことで黒髭人形が飛び出す仕組みである。樽に相当するDNA(樽DNA)と人形に相当するDNA（人形DNA）をミスマッチさせた相補鎖の状態で結合させておく。その後、樽DNAとマッチするDNA（剣DNA）を混合させる。そうすること、人形DNAと剣DNAが入れ替わり、人形DNAが飛んでいくように見える。このような、機構で目視検出可能である選択的に物資を放出するデバイスの作製を試みた。（図：DNA origamiのみで作ろうとしたものをアニメーションで表現）<br>
&nbsp;Pop-up Pirate is one of the famous Japanese toy .Players bury a sward into the barrel. When the player bury into the specific position, the doll will be fly away. The barrel is constructed with the single strand DNA binding porous silica particle. Complemental DNA is a roll of the doll. But there are same mismatch between barrel and doll DNAs. Fully complement single strand DNA in a sword. Firstly, doll DNA is mixed with the barrel. Then doll DNA is bound to the barrel with complementally base pairing. Next, sward DNA is added to the solution. The doll DNA would be released firm the barrel because affinity of the sward DNA and the barrel .DNA is strange than that of the doll and the barrel DNAs. At looks like the doll fly away from the barrel.  <br>
&nbsp; しかしながら、全ての部品をDNA origamiで作製することは観察面、コスト面を考慮すると現実的ではない。そこで、シリカ粒子にDNAを結合させることにより、コストを削減することができ、また観察を容易にすることができる。また不完全な塩基対から完全な塩基対に置き換わったか顕微鏡で確認することは難しいので、2種類の蛍光分子を用いて確認できるようにした。<br></p>
&nbsp;In order to detect the different state of DNA binding, FRET system is used. The barrel DNA is labeled with fluorescent bye TAMRA, and the barrel DNAs are bound, TAMURA was located near the FITC.  <br></p>




&nbsp; 蛍光分子は蛍光顕微鏡の単一レーザーの波長が488nmであるため、励起波長が488nmであるFITC（ハイパーリンク）とFITCの蛍光波長で励起するTAMRA（ハイパーリンク）を用いた。<br>
&nbsp; Therefore, FITC is excited by the 488nm laser light and TAMRA is excited by the fluorescent of FITC. <br>




&nbsp; 樽DNAとミスマッチな塩基対を有する人形DNAの末端に緑色の蛍光分子を修飾し、樽DNAの末端に緑色の波長を吸収して赤く蛍光する分子を修飾する。樽DNAに剣DNAを混合する。人形DNAの時は二つの蛍光分子の相互作用により赤く蛍光し、人形ＤＮＡが先ほどの剣DNAと置き換わることで消光する。検出される光強度が変化することで数値として明確に放出されたか表すことができる<br clear="all"><br>
&nbsp;Then we observed the red fluorescent around the silica particles under the confocal fluorescent microscope. When we added the sward DNA and FITC labeled doll DNA is released from the TAMRA labeled barrel DNA .We could not observed any fluorescent around  <br clear="all"><br>
(図：完成形をアニメーションで表現)
(図：完成形をアニメーションで表現)
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Revision as of 11:37, 29 August 2014

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<div id="header"><h1><p style="background:#CCFFFF; color:purple;"><font face=cursive size="6"><B> Team FIT </font> </B></p></a></h1></div>





<table> <tr align="center"> <td cellspacing="10" cellpadding="10" width="1055" width="180" height="60" bgcolor="#BAD3FF"><a href="http://openwetware.org/wiki/Biomod/2014/Fukuoka"><B><font face=cursive color="#003366" size="3">Top</font></B></a></td> <td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Introduction.html"><B><font face=cursive color="#003366" size="3">Introduction</font></B></a></td> <td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Approach and Goals.html"><B><font face=cursive color="#FF6633" size="3" >Approach and Goals</font></B></a></td> <td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Method.html"><B><font face=cursive color="#003366" size="3">Method</font></B></a></td> <td cellspacing="10" cellpadding="10" width="1300" width="180" bgcolor="#BAD3FF"><a href="fit_Results and Discussion.html"><B><font face=cursive color="#003366" size="3">Results and Discussion</font></B></a></td> <td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Member.html"><B><font face=cursive color="#003366" size="3">Member</font></B></a></td> <td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Sponsor.html"><B><font face=cursive color="#003366" size="3">Sponsor</font></B></a></td> </tr></table>



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<h2 p style="background:#FFFFFF; color:#003366;"><font face=cursive size="5"><B> Approach and Goals </font></B></p></h2>


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<h3><p>&nbsp;We paid attention to complementary base pair of DNA Single strand DNA of two can be connected even if I make a mistake, and base pair matches it to some extent. The fewer and fewer mismatched base pair, Cohesion is high. We tried to construct the Pop-up Pirate using DNA and porous silica particle. <br>

<p> <table align="left"> <td><div style="width: 330px;"><img alt="" src="/images/c/c7/Dnakurohige.gif" width="320" height="240" border="0" / ><p>(*^_^*)</p><div></td><tr> <td><div style="width: 330px;"><img alt="" src="/images/b/b7/Fitfiani.gif" width="320" height="240" border="0" / ><p>(*^_^*)</p><div></td></table>

&nbsp;Pop-up Pirate is one of the famous Japanese toy .Players bury a sward into the barrel. When the player bury into the specific position, the doll will be fly away. The barrel is constructed with the single strand DNA binding porous silica particle. Complemental DNA is a roll of the doll. But there are same mismatch between barrel and doll DNAs. Fully complement single strand DNA in a sword. Firstly, doll DNA is mixed with the barrel. Then doll DNA is bound to the barrel with complementally base pairing. Next, sward DNA is added to the solution. The doll DNA would be released firm the barrel because affinity of the sward DNA and the barrel .DNA is strange than that of the doll and the barrel DNAs. At looks like the doll fly away from the barrel. <br> &nbsp;In order to detect the different state of DNA binding, FRET system is used. The barrel DNA is labeled with fluorescent bye TAMRA, and the barrel DNAs are bound, TAMURA was located near the FITC. <br></p>


&nbsp; Therefore, FITC is excited by the 488nm laser light and TAMRA is excited by the fluorescent of FITC. <br>


&nbsp;Then we observed the red fluorescent around the silica particles under the confocal fluorescent microscope. When we added the sward DNA and FITC labeled doll DNA is released from the TAMRA labeled barrel DNA .We could not observed any fluorescent around <br clear="all"><br> (図：完成形をアニメーションで表現) </p></h3>




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