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− | <a href="#!" onclick="toggleHeight3(this, | + | <a href="#!" onclick="toggleHeight3(this, 6000); return false" |
style="font-family:'Acme', sans-serif;font-size:30px;color:#205e1a;height: 30px;"> | style="font-family:'Acme', sans-serif;font-size:30px;color:#205e1a;height: 30px;"> | ||
Experiments | Experiments | ||
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<h4>NrtA expression</h4> | <h4>NrtA expression</h4> | ||
<p> We successfully transformed NrtA gene from cyanobacteria <i>Synechocystis</i> sp. PCC 6803 to <i>E.coli</i>.</p> | <p> We successfully transformed NrtA gene from cyanobacteria <i>Synechocystis</i> sp. PCC 6803 to <i>E.coli</i>.</p> | ||
− | <center><img src="https://static.igem.org/mediawiki/2017/d/da/T--NYMU-Taipei--NS_NrtA_RE.png" style="width: | + | <center><img src="https://static.igem.org/mediawiki/2017/d/da/T--NYMU-Taipei--NS_NrtA_RE.png" style="width:60%;"></center> |
<p> This figure is restriction enzyme check electrophoresis result of NrtA construct. | <p> This figure is restriction enzyme check electrophoresis result of NrtA construct. | ||
We use XhoI and HindIII to digest the plasmid, and the expected length is 892bp, 1169bp, 1540bp. | We use XhoI and HindIII to digest the plasmid, and the expected length is 892bp, 1169bp, 1540bp. | ||
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<h4>endolysin construct</h4> | <h4>endolysin construct</h4> | ||
<p> We successfully constructed J23106-B0034-Endolysin-B0010-B0012. The endolysin in this part is from iGEM released part, BBa_K112806, which is endolysin from enterobacteria phage T4. Besides endolysin, in this composite part, we choose BBa_J23106 as a constitutive promoter, BBa_B0034 as ribosome binding site, BBa_B0010 and BBa_B0012 as double terminator, all of which are widely used parts in iGEM.</p> | <p> We successfully constructed J23106-B0034-Endolysin-B0010-B0012. The endolysin in this part is from iGEM released part, BBa_K112806, which is endolysin from enterobacteria phage T4. Besides endolysin, in this composite part, we choose BBa_J23106 as a constitutive promoter, BBa_B0034 as ribosome binding site, BBa_B0010 and BBa_B0012 as double terminator, all of which are widely used parts in iGEM.</p> | ||
− | <div style="width: | + | <div style="width:55%;margin:auto;"> |
<img src="https://static.igem.org/mediawiki/2017/c/c4/T--NYMU-Taipei--NS_endolysin_PCR.png" style="width:95%;"> | <img src="https://static.igem.org/mediawiki/2017/c/c4/T--NYMU-Taipei--NS_endolysin_PCR.png" style="width:95%;"> | ||
<p>This figure is electrophoresis result of endolysin PCR product. | <p>This figure is electrophoresis result of endolysin PCR product. | ||
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<h4>holin construct</h4> | <h4>holin construct</h4> | ||
<p> We successfully constructed R0010-B0034-Holin-B0010-B0012. To control the precise suicide timing, we choose a lactose-induced promoter, BBa_R0010, to regulate this suicide mechanism. Besides holin and inducible promoter, in this composite part, we also choose BBa_B0034 as ribosome binding site, and BBa_B0010 and BBa_B0012 as double terminator.</p> | <p> We successfully constructed R0010-B0034-Holin-B0010-B0012. To control the precise suicide timing, we choose a lactose-induced promoter, BBa_R0010, to regulate this suicide mechanism. Besides holin and inducible promoter, in this composite part, we also choose BBa_B0034 as ribosome binding site, and BBa_B0010 and BBa_B0012 as double terminator.</p> | ||
− | <div style="width: | + | <div style="width:55%;margin:auto;"> |
<img src="https://static.igem.org/mediawiki/2017/9/9a/T--NYMU-Taipei--NS_holin_PCR.png" style="width:95%;"> | <img src="https://static.igem.org/mediawiki/2017/9/9a/T--NYMU-Taipei--NS_holin_PCR.png" style="width:95%;"> | ||
<p>This figure is electrophoresis result of holin backbone PCR product. | <p>This figure is electrophoresis result of holin backbone PCR product. | ||
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<h4>endolysin-holin construct</h4> | <h4>endolysin-holin construct</h4> | ||
<p> We successfully constructed R0010-B0034-Holin-B0010-B0012-J23106-B0034-Endolysin-B0010-B0012. We combined holin and endolysin for suicide mechanism. In this composite part, holin functions as an important regulation role.</p> | <p> We successfully constructed R0010-B0034-Holin-B0010-B0012-J23106-B0034-Endolysin-B0010-B0012. We combined holin and endolysin for suicide mechanism. In this composite part, holin functions as an important regulation role.</p> | ||
− | <div style="width: | + | <div style="width:55%;margin:auto;"> |
<img src="https://static.igem.org/mediawiki/parts/e/e2/T--NYMU-Taipei%E2%80%94Holin-Endolysin-REcheck.png" style="width:50%;"> | <img src="https://static.igem.org/mediawiki/parts/e/e2/T--NYMU-Taipei%E2%80%94Holin-Endolysin-REcheck.png" style="width:50%;"> | ||
<p>This is restriction enzyme check electrophoresis result of holin-endolysin construct. | <p>This is restriction enzyme check electrophoresis result of holin-endolysin construct. | ||
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</div> | </div> | ||
+ | <h4></h4> | ||
<h4>endolysin-holin-NrtA construct</h4> | <h4>endolysin-holin-NrtA construct</h4> | ||
+ | <p> We successfully constructed R0010-B0034-Holin-B0010-B0012-J23106-B0034-Endolysin-B0010-B0012-J23118-B0034-NrtA-B0015. This part combined holin, endolysin, and NrtA and had nitrate-capturing function and suicide function.</p> | ||
+ | <div style="width:55%;margin:auto;"> | ||
+ | <img src="https://static.igem.org/mediawiki/parts/b/b0/Holin-Endolysin-NrtA_REcheck.png" style="width:50%;"> | ||
+ | <p>This figure is restriction enzyme check electrophoresis result of endolysin-holin-NrtA construct. | ||
+ | We use XmaI to digest sample 1~3. The result shows that 2 and 3 are right. M represents 1 kb marker. | ||
+ | </p> | ||
+ | </div> | ||
+ | |||
<p>*See our parts: <a href="https://2017.igem.org/Team:NYMU-Taipei/Parts" target="_blank">click</a></p> | <p>*See our parts: <a href="https://2017.igem.org/Team:NYMU-Taipei/Parts" target="_blank">click</a></p> | ||
<p>*See our experiments protocols: <a href="https://2017.igem.org/Team:NYMU-Taipei/Notebook" target="_blank">click</a></p> | <p>*See our experiments protocols: <a href="https://2017.igem.org/Team:NYMU-Taipei/Notebook" target="_blank">click</a></p> | ||
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<br> | <br> | ||
<center> | <center> | ||
− | <a href="#!" onclick="toggleHeight3(this, | + | <a href="#!" onclick="toggleHeight3(this, 6000);" style='color:#56914a'> |
click to close | click to close | ||
</a> | </a> |
Revision as of 20:02, 28 October 2017