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Team:Paris Bettencourt/Notebook/Drug Screening/Friday 5th July.html - Revision history
2024-03-29T10:40:29Z
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http://2013.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Notebook/Drug_Screening/Friday_5th_July.html&diff=69090&oldid=prev
Yzegman at 09:43, 13 August 2013
2013-08-13T09:43:39Z
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Place your twit here</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Planning transformations for the next week</ins></div></td></tr>
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Yzegman
http://2013.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Notebook/Drug_Screening/Friday_5th_July.html&diff=69083&oldid=prev
Yzegman at 09:41, 13 August 2013
2013-08-13T09:41:12Z
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Drug_Screen" target="_blank" class="tbnotelogo DSlogo"> ASDF </a></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Drug_Screen" target="_blank" class="tbnotelogo DSlogo"> ASDF </a></div></td></tr>
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Yzegman
http://2013.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Notebook/Drug_Screening/Friday_5th_July.html&diff=69080&oldid=prev
Yzegman at 09:40, 13 August 2013
2013-08-13T09:40:34Z
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">These</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">These</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>transformations <del class="diffchange diffchange-inline">were </del>done by using two strains of competent cells</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>transformations <ins class="diffchange diffchange-inline">will be </ins>done by using two strains of competent cells</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>prepared using Protocol 2 :</SPAN></FONT></FONT></FONT></P></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>prepared using Protocol 2 :</SPAN></FONT></FONT></FONT></P></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>transformations <del class="diffchange diffchange-inline">were </del>done by heat shock using protocol 1.</SPAN></FONT></FONT></FONT></P></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>transformations <ins class="diffchange diffchange-inline">will be </ins>done by heat shock using protocol 1.</SPAN></FONT></FONT></FONT></P></div></td></tr>
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Yzegman
http://2013.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Notebook/Drug_Screening/Friday_5th_July.html&diff=67953&oldid=prev
Yzegman at 15:23, 12 August 2013
2013-08-12T15:23:24Z
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Yzegman
http://2013.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Notebook/Drug_Screening/Friday_5th_July.html&diff=67947&oldid=prev
Yzegman at 15:22, 12 August 2013
2013-08-12T15:22:28Z
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>FNR gene (pD004), was found not to be resistant as well. </SPAN></FONT></FONT></FONT></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>FNR gene (pD004), was found not to be resistant as well. </SPAN></FONT></FONT></FONT></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <P STYLE="margin-top: 0.08in; margin-bottom: 0.08in"><IMG SRC="https://static.igem.org/mediawiki/2013/thumb/9/91/Wishplate.jpg/800px-Wishplate.jpg" NAME="graphics2" ALIGN=LEFT WIDTH=626 HEIGHT=462 BORDER=0><BR CLEAR=LEFT><FONT SIZE=3><I>Illustration</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <P STYLE="margin-top: 0.08in; margin-bottom: 0.08in"><IMG SRC="https://static.igem.org/mediawiki/2013/thumb/9/91/Wishplate.jpg/800px-Wishplate.jpg" NAME="graphics2" ALIGN=LEFT WIDTH=626 HEIGHT=462 BORDER=0><BR CLEAR=LEFT><FONT SIZE=3><I>Illustration</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> 1: Strains will be grown on a minimal media M9 plate, supplemented</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> 1: Strains will be grown on a minimal media M9 plate, supplemented</div></td></tr>
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Yzegman
http://2013.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Notebook/Drug_Screening/Friday_5th_July.html&diff=67936&oldid=prev
Yzegman at 15:18, 12 August 2013
2013-08-12T15:18:24Z
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <P STYLE="margin-top: 0.08in; margin-bottom: 0.08in"><IMG SRC="https://static.igem.org/mediawiki/2013/thumb/9/91/Wishplate.jpg/800px-Wishplate.jpg" NAME="graphics2" ALIGN=<del class="diffchange diffchange-inline">BOTTOM </del>WIDTH=626 HEIGHT=462 BORDER=0><BR CLEAR=LEFT><FONT SIZE=3><I>Illustration</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <P STYLE="margin-top: 0.08in; margin-bottom: 0.08in"><IMG SRC="https://static.igem.org/mediawiki/2013/thumb/9/91/Wishplate.jpg/800px-Wishplate.jpg" NAME="graphics2" ALIGN=<ins class="diffchange diffchange-inline">LEFT </ins>WIDTH=626 HEIGHT=462 BORDER=0><BR CLEAR=LEFT><FONT SIZE=3><I>Illustration</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> 1: Strains will be grown on a minimal media M9 plate, supplemented</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> 1: Strains will be grown on a minimal media M9 plate, supplemented</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> with all amino acids aside from the sulfur containing ones; plates</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> with all amino acids aside from the sulfur containing ones; plates</div></td></tr>
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Yzegman
http://2013.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Notebook/Drug_Screening/Friday_5th_July.html&diff=67918&oldid=prev
Yzegman at 15:14, 12 August 2013
2013-08-12T15:14:07Z
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <P STYLE="margin-top: 0.08in; margin-bottom: 0.08in"><IMG SRC="https://static.igem.org/mediawiki/2013/thumb/9/91/Wishplate.jpg/800px-Wishplate.jpg" NAME="graphics2" ALIGN=<del class="diffchange diffchange-inline">LEFT </del>WIDTH=<del class="diffchange diffchange-inline">100% </del>BORDER=0><BR CLEAR=LEFT><FONT SIZE=3><I>Illustration</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <P STYLE="margin-top: 0.08in; margin-bottom: 0.08in"><IMG SRC="https://static.igem.org/mediawiki/2013/thumb/9/91/Wishplate.jpg/800px-Wishplate.jpg" NAME="graphics2" ALIGN=<ins class="diffchange diffchange-inline">BOTTOM </ins>WIDTH=<ins class="diffchange diffchange-inline">626 HEIGHT=462 </ins>BORDER=0><BR CLEAR=LEFT><FONT SIZE=3><I>Illustration</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> 1: Strains will be grown on a minimal media M9 plate, supplemented</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> 1: Strains will be grown on a minimal media M9 plate, supplemented</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> with all amino acids aside from the sulfur containing ones; plates</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> with all amino acids aside from the sulfur containing ones; plates</div></td></tr>
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Yzegman
http://2013.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Notebook/Drug_Screening/Friday_5th_July.html&diff=67897&oldid=prev
Yzegman at 15:09, 12 August 2013
2013-08-12T15:09:38Z
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><P STYLE="margin-bottom: 0in; line-height: 114%"><SPAN ID="Frame1" DIR="LTR" STYLE="float: left; width: 4.51in; height: 3.55in; border: none; padding: 0in; background: #ffffff"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><P STYLE="margin-bottom: 0in; line-height: 114%"><SPAN ID="Frame1" DIR="LTR" STYLE="float: left; width: 4.51in; height: 3.55in; border: none; padding: 0in; background: #ffffff"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <P STYLE="margin-top: 0.08in; margin-bottom: 0.08in"><IMG SRC="../<del class="diffchange diffchange-inline">Downloads</del>/Wishplate.jpg" NAME="graphics2" ALIGN=LEFT WIDTH=100% BORDER=0><BR CLEAR=LEFT><FONT SIZE=3><I>Illustration</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <P STYLE="margin-top: 0.08in; margin-bottom: 0.08in"><IMG SRC="<ins class="diffchange diffchange-inline">https://static</ins>.<ins class="diffchange diffchange-inline">igem</ins>.<ins class="diffchange diffchange-inline">org</ins>/<ins class="diffchange diffchange-inline">mediawiki/2013/thumb/9/91/Wishplate.jpg</ins>/<ins class="diffchange diffchange-inline">800px-</ins>Wishplate.jpg" NAME="graphics2" ALIGN=LEFT WIDTH=100% BORDER=0><BR CLEAR=LEFT><FONT SIZE=3><I>Illustration</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> 1: Strains will be grown on a minimal media M9 plate, supplemented</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> 1: Strains will be grown on a minimal media M9 plate, supplemented</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> with all amino acids aside from the sulfur containing ones; plates</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> with all amino acids aside from the sulfur containing ones; plates</div></td></tr>
</table>
Yzegman
http://2013.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Notebook/Drug_Screening/Friday_5th_July.html&diff=67885&oldid=prev
Yzegman: Created page with "<html> <div class ="tbnote"> <h2>Drug Screening</h2> <a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Drug_Screen" target="_blank" class="tbnotelogo DSlogo"> ASDF </a..."
2013-08-12T15:06:58Z
<p>Created page with "<html> <div class ="tbnote"> <h2>Drug Screening</h2> <a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Drug_Screen" target="_blank" class="tbnotelogo DSlogo"> ASDF </a..."</p>
<p><b>New page</b></p><div><html><br />
<div class ="tbnote"><br />
<h2>Drug Screening</h2><br />
<a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Drug_Screen" target="_blank" class="tbnotelogo DSlogo"> ASDF </a><br />
<div style="clear: both;"></div><br />
<h3>Day Number<sup>suffix</sup> Month</h3><br />
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<P STYLE="margin-bottom: 0in; line-height: 114%"><A NAME="docs-internal-guid-1ecd545a-72a7-d313-3414-d51c38553976"></A><br />
<FONT COLOR="#222222"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"><SPAN STYLE="background: #ffffff">We<br />
decided to partly reproduce Figure 3C from:<br />
</SPAN></SPAN></SPAN></FONT></FONT></SPAN></FONT><A HREF="http://www.jbioleng.org/content/5/1/7"><FONT COLOR="#005400"><SPAN STYLE="text-decoration: none"><FONT FACE="Verdana"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="font-style: normal"><U><SPAN STYLE="font-weight: normal"><SPAN STYLE="background: #ffffff">http://www.jbioleng.org/content/5/1/7</SPAN></SPAN></U></SPAN></FONT></FONT></SPAN></FONT></A><FONT COLOR="#222222"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"><SPAN STYLE="background: #ffffff">:</SPAN></SPAN></SPAN></FONT></FONT></SPAN></FONT></P><br />
<P STYLE="font-weight: normal"><FONT SIZE=2 STYLE="font-size: 11pt"><BR><IMG SRC="https://lh5.googleusercontent.com/EKwdYjSSLfO_91W7fgqGLKIPkkGjAMWWrfB0GcvR5o1wlQJsBaa5cqajDhreNfAi5ZS1cLaMgKrws8sn6VhEGAAgIfEajdqALZCCNgrDG9NSD0AoEV_kLXdO" NAME="graphics1" ALIGN=BOTTOM WIDTH=626 HEIGHT=462 BORDER=0></FONT></P><br />
<P STYLE="margin-bottom: 0in; font-style: normal; font-weight: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#222222"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">So,<br />
we would pursue to grow the following strains on a minimal media M9<br />
plate, supplemented with all amino acids</SPAN></FONT></FONT></FONT></P><br />
<P STYLE="margin-bottom: 0in; font-style: normal; font-weight: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#222222"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">aside<br />
from the sulfur containing ones, and sulfite ans the only sulfur<br />
source:</SPAN></FONT></FONT></FONT></P><br />
<P STYLE="margin-bottom: 0in; font-style: normal; font-weight: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#222222"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">We<br />
already thought we have all of the strains we need, however, an<br />
overnight culture was launched of a few single colonies from this<br />
double plasmid strain (sD001+pD004+pD005, which supposedly had all 3<br />
genes SIR FNR and Fdx) and it didn't grow. </SPAN></FONT></FONT></FONT><br />
</P><br />
<P STYLE="margin-bottom: 0in; font-weight: normal; line-height: 114%"><br />
<FONT SIZE=2 STYLE="font-size: 11pt"><FONT COLOR="#222222"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><SPAN STYLE="background: #ffffff">This<br />
most likely occurred because the strains didn't have </SPAN></SPAN></FONT></SPAN></FONT><FONT COLOR="#222222"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><I><SPAN STYLE="background: #ffffff">Spec<br />
</SPAN></I></FONT></SPAN></FONT><FONT COLOR="#222222"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><SPAN STYLE="background: #ffffff">antibiotics<br />
resistance, as the other spec resistant strain containing</SPAN></SPAN></FONT></SPAN></FONT></FONT></P><br />
<P STYLE="margin-bottom: 0in; font-style: normal; font-weight: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#222222"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">the<br />
FNR gene (pD004), was found not to be resistant as well. </SPAN></FONT></FONT></FONT><br />
</P><br />
<P STYLE="margin-bottom: 0in; line-height: 114%"><SPAN ID="Frame1" DIR="LTR" STYLE="float: left; width: 4.51in; height: 3.55in; border: none; padding: 0in; background: #ffffff"><br />
<P STYLE="margin-top: 0.08in; margin-bottom: 0.08in"><IMG SRC="../Downloads/Wishplate.jpg" NAME="graphics2" ALIGN=LEFT WIDTH=100% BORDER=0><BR CLEAR=LEFT><FONT SIZE=3><I>Illustration<br />
1: Strains will be grown on a minimal media M9 plate, supplemented<br />
with all amino acids aside from the sulfur containing ones; plates<br />
will have increased glucose, and sulfite as the only sulfur source.<br />
Growth is expected only for WT and the strain containing all three<br />
alternative genes of sulfur reduction.</I></FONT></P><br />
</SPAN><BR CLEAR=LEFT><BR><br />
</P><br />
<P STYLE="margin-bottom: 0in; line-height: 114%"><BR><br />
</P><br />
<P STYLE="margin-bottom: 0in; font-style: normal; font-weight: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#222222"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">Therefore,<br />
we would &nbsp;re-make: </SPAN></FONT></FONT></FONT><br />
</P><br />
<UL><br />
<LI><P STYLE="margin-right: 0.15in; margin-bottom: 0in; background: transparent; font-style: normal; font-weight: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><FONT COLOR="#222222"><B><SPAN STYLE="background: #ffffff">sD001+pD004+pD005</SPAN></B></FONT><FONT COLOR="#222222"><SPAN STYLE="background: #ffffff"><br />
E.coli:</SPAN></FONT><FONT COLOR="#ff9900"><SPAN STYLE="background: #ffffff">ΔcysI,<br />
Δfpr, ΔydbK</SPAN></FONT><SPAN STYLE="background: #ffffff">::<br />
</SPAN><FONT COLOR="#0000ff"><B><SPAN STYLE="background: #ffffff">soFD</SPAN></B></FONT><SPAN STYLE="background: #ffffff">,<br />
</SPAN><FONT COLOR="#ff0000"><B><SPAN STYLE="background: #ffffff">zmSIR<br />
</SPAN></B></FONT><SPAN STYLE="background: #ffffff">;Chloramphenicol<br />
,</SPAN><FONT COLOR="#ff00ff"><B><SPAN STYLE="background: #ffffff">zmFNR</SPAN></B></FONT><FONT COLOR="#222222"><SPAN STYLE="background: #ffffff">;<br />
Spectinomycin- </SPAN></FONT><FONT COLOR="#222222"><U><SPAN STYLE="background: #ffffff">with<br />
all three genes.</SPAN></U></FONT></FONT></FONT></FONT></P><br />
<LI><P STYLE="margin-right: 0.15in; margin-bottom: 0in; background: transparent; font-style: normal; font-weight: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><FONT COLOR="#222222"><U><B><SPAN STYLE="background: #ffffff">sD001+pD004:<br />
</SPAN></B></U></FONT><FONT COLOR="#222222"><SPAN STYLE="background: #ffffff">E.coli:</SPAN></FONT><FONT COLOR="#ff9900"><SPAN STYLE="background: #ffffff">ΔcysI,<br />
Δfpr, ΔydbK</SPAN></FONT><SPAN STYLE="background: #ffffff">::<br />
</SPAN><FONT COLOR="#ff00ff"><B><SPAN STYLE="background: #ffffff">zmFNR</SPAN></B></FONT><FONT COLOR="#222222"><SPAN STYLE="background: #ffffff">;<br />
Spectinomycin</SPAN></FONT></FONT></FONT></FONT></P><br />
</UL><br />
<P><BR><BR><br />
</P><br />
<P STYLE="margin-right: 0.15in; margin-bottom: 0in; font-style: normal; font-weight: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#222222"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">and<br />
make</SPAN></FONT></FONT></FONT></P><br />
<P STYLE="margin-right: 0.15in; margin-bottom: 0in; line-height: 114%"><br />
<BR><br />
</P><br />
<UL><br />
<LI><P STYLE="margin-right: 0.15in; margin-bottom: 0in; background: transparent; line-height: 114%"><br />
<FONT SIZE=2 STYLE="font-size: 11pt"><FONT COLOR="#222222"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><B><SPAN STYLE="background: #ffffff">sD001+pD004+pD006</SPAN></B></SPAN></FONT></SPAN></FONT><FONT COLOR="#222222"><SPAN STYLE="text-decoration: none"><SPAN STYLE="background: #ffffff"><br />
</SPAN></SPAN></FONT><FONT COLOR="#222222"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"><SPAN STYLE="background: #ffffff">E.coli:</SPAN></SPAN></SPAN></FONT></SPAN></FONT><FONT COLOR="#ff9900"><SPAN STYLE="text-decoration: none"><SPAN STYLE="background: #ffffff"><br />
</SPAN></SPAN></FONT><FONT COLOR="#ff9900"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"><SPAN STYLE="background: #ffffff">ΔcysI,<br />
Δfpr, ΔydbK</SPAN></SPAN></SPAN></FONT></SPAN></FONT><FONT COLOR="#000000"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"><SPAN STYLE="background: #ffffff">::</SPAN></SPAN></SPAN></FONT></SPAN></FONT><FONT COLOR="#ff00ff"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><B><SPAN STYLE="background: #ffffff">zmFNR</SPAN></B></SPAN></FONT></SPAN></FONT><FONT COLOR="#000000"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"><SPAN STYLE="background: #ffffff">;<br />
Spectinomycin, </SPAN></SPAN></SPAN></FONT></SPAN></FONT><FONT COLOR="#ff0000"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><B><SPAN STYLE="background: #ffffff">zmSIR</SPAN></B></SPAN></FONT></SPAN></FONT><FONT COLOR="#000000"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"><SPAN STYLE="background: #ffffff">;<br />
Chloramphenicol</SPAN></SPAN></SPAN></FONT></SPAN></FONT></FONT></P><br />
</UL><br />
<P STYLE="margin-right: 0.15in; margin-bottom: 0in; background: transparent; line-height: 114%"><br />
<BR><br />
</P><br />
<P STYLE="margin-right: 0.15in; margin-bottom: 0in; background: transparent; font-style: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">These<br />
transformations were done by using two strains of competent cells<br />
prepared using Protocol 2 :</SPAN></FONT></FONT></FONT></P><br />
<UL><br />
<LI><P STYLE="margin-bottom: 0in; background: transparent; font-style: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">sD001<br />
competent cells were transformed with:</SPAN></FONT></FONT></FONT></P><br />
<UL><br />
<LI><P STYLE="margin-bottom: 0in; background: transparent; font-style: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">pD004</SPAN></FONT></FONT></FONT></P><br />
<LI><P STYLE="margin-bottom: 0in; background: transparent; font-style: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">Double<br />
transformation: pD005 +pD004</SPAN></FONT></FONT></FONT></P><br />
</UL><br />
<LI><P STYLE="margin-bottom: 0in; background: transparent; font-style: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">sD007<br />
(sD001+pD005) competent cells were transformed with:</SPAN></FONT></FONT></FONT></P><br />
<UL><br />
<LI><P STYLE="margin-bottom: 0in; background: transparent; font-style: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">pD004</SPAN></FONT></FONT></FONT></P><br />
</UL><br />
</UL><br />
<P STYLE="margin-bottom: 0in; background: transparent; font-style: normal; line-height: 114%; text-decoration: none"><br />
<FONT COLOR="#000000"><FONT FACE="Arial"><FONT SIZE=2 STYLE="font-size: 11pt"><SPAN STYLE="background: #ffffff">These<br />
transformations were done by heat shock using protocol 1.</SPAN></FONT></FONT></FONT></P><br />
<P STYLE="font-weight: normal"><FONT SIZE=2 STYLE="font-size: 11pt"><BR></FONT><BR><BR><br />
</P><br />
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