Modular Synthetic Receptor System

From 2008.igem.org

(Difference between revisions)
m
Line 15: Line 15:
<h2>'''Discussion:'''</h2>
<h2>'''Discussion:'''</h2>
<h2>'''Literature:'''</h2>
<h2>'''Literature:'''</h2>
 +
 +
<h2>'''Tabelle fuer Kathrin'''</h2>
 +
<table class="MsoTableGrid"
 +
style="border: medium none ; border-collapse: collapse;" border="1"
 +
cellpadding="0" cellspacing="0" width="679">
 +
    <tr style="">
 +
      <td
 +
style="border: 1pt solid windowtext; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Step 1</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: solid solid solid none; border-color: windowtext windowtext windowtext -moz-use-text-color; border-width: 1pt 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Vector</span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">EcoRI + PstI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
      <td
 +
style="border-style: solid solid solid none; border-color: windowtext windowtext windowtext -moz-use-text-color; border-width: 1pt 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Insert
 +
      </span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">EcoRI + PstI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">&nbsp;</span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">BBa-J52017<b
 +
style=""><span style="color: purple;"></span></b></span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style="">CMV</b>-promotor</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border: 1pt solid windowtext; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Step
 +
2</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Vector</span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">AgeI+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Insert
 +
      </span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">NgoMIV+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">&nbsp;</span></p>
 +
      </td>
 +
      <td rowspan="2"
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">pMA-BBFR
 +
_<b style=""><span style="color: rgb(255, 102, 0);"> </span><span
 +
style="color: blue;">SPLIT-Linker</span></b></span></p>
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: rgb(255, 102, 0);" lang="EN-GB">C</span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-YFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: rgb(255, 102, 0);" lang="EN-GB">C</span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-CFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border: 1pt solid windowtext; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Step
 +
3</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Vector</span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">AgeI+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Insert
 +
      </span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">NgoMIV+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">&nbsp;</span></p>
 +
      </td>
 +
      <td rowspan="8"
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">pMA-BBFR
 +
_<b style=""><span style="color: maroon;">egfR</span>-<span
 +
style="color: maroon;">Tm</span></b></span></p>
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_ <span style="color: rgb(255, 102, 0);"><span style="">&nbsp;</span>N</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-&#946;-Lactamase</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_ <span style="color: rgb(255, 102, 0);"><span style="">&nbsp;</span>C</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-&#946;-Lactamase</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_ <span style="color: blue;">SPLIT-Linker</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_<b style=""><span
 +
style="color: rgb(255, 102, 0);"> C</span></b>-YFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_ <span style="color: rgb(255, 102, 0);">N</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-YFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_ <span style="color: blue;">SPLIT-Linker</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_<b style=""><span
 +
style="color: rgb(255, 102, 0);"> C</span></b>-CFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_ <span style="color: rgb(255, 102, 0);">N</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-CFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: rgb(255, 102, 0);"> </span></b>BB0<b
 +
style=""><span style="color: rgb(255, 102, 0);">58 </span></b>(Luciferase)</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="height: 3.5pt;">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt; height: 3.5pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt; height: 3.5pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: rgb(255, 102, 0);"> </span></b>BB0<b
 +
style=""><span style="color: rgb(255, 102, 0);">57<span style="">&nbsp; </span></span></b>(Luciferase)</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border: 1pt solid windowtext; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Step
 +
4</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Vector</span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">AgeI+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Insert
 +
      </span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">NgoMIV+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">&nbsp;</span></p>
 +
      </td>
 +
      <td rowspan="2"
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">pMA-BBFR
 +
_<b style=""><span style="color: purple;">SP</span></b></span></p>
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: rgb(51, 153, 102);" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: rgb(51, 102, 255);" lang="EN-GB">_</span></b><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 51, 0);"
 +
lang="EN-GB">scFv-anti-NIP</span></b><span style="font-size: 10pt;"
 +
lang="EN-GB"></span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">&nbsp;</span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span><span
 +
style="color: rgb(51, 102, 255);"></span></b></span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border: 1pt solid windowtext; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Step
 +
5</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Vector</span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">AgeI+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Insert
 +
      </span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">NgoMIV+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">&nbsp;</span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">pMA-BBFR
 +
_<b style=""><span style="color: purple;">SP</span></b>_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(153, 51, 0);">scFv-anti-NIP</span></b></span></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt; color: black;"
 +
lang="EN-GB">and</span><span style="font-size: 11pt; color: black;"
 +
lang="EN-GB"></span></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">pMA-BBFR-+<b
 +
style=""><span style="color: purple;">SP</span></b>_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span><span
 +
style="color: rgb(51, 153, 102);"></span></b></span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style="">GGGS-linker</b> (produced by Klenow fill in)</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border: 1pt solid windowtext; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Step
 +
6</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Vector</span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">AgeI+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Insert
 +
      </span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion: </span><span
 +
style="font-size: 8.5pt;">NgoMIV+SpeI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">&nbsp;</span></p>
 +
      </td>
 +
      <td rowspan="8"
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">pMA-BBFR
 +
_<b style=""><span style="color: purple;">SP</span></b>_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(153, 51, 0);">scFv-anti-NIP</span></b> _ <b style="">GGGS-Li</b></span></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">and</span></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">pMA-BBFR
 +
_<b style=""><span style="color: purple;"> SP</span></b>_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span></b> _</span></p>
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">GGGS-Li</span></b><span style="font-size: 10pt;"
 +
lang="EN-GB"></span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"> <span
 +
lang="EN-GB">_ <span style="color: rgb(255, 102, 0);"><span style="">&nbsp;</span>N</span></span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-&#946;-Lactamase</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"> <span
 +
lang="EN-GB">_ <span style="color: rgb(255, 102, 0);"><span style="">&nbsp;</span>C</span></span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-&#946;-Lactamase</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"> <span
 +
lang="EN-GB">_ SPLIT-Linker</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_<b style=""><span
 +
style="color: rgb(255, 102, 0);"> C</span></b>-YFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"> <span
 +
lang="EN-GB">_ <span style="color: rgb(255, 102, 0);">N</span></span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-YFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"> <span
 +
lang="EN-GB">_ SPLIT-Linker</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_<b style=""><span
 +
style="color: rgb(255, 102, 0);"> C</span></b>-CFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"> <span
 +
lang="EN-GB">_ <span style="color: rgb(255, 102, 0);">N</span></span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-CFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: maroon;" lang="EN-GB">_</span></b><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><span style="font-size: 10pt;" lang="EN-GB"> _<b
 +
style=""><span style="color: rgb(255, 102, 0);"> </span></b>BB0<b
 +
style=""><span style="color: rgb(255, 102, 0);">58 </span></b>(Luciferase)</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><span style="font-size: 10pt;" lang="EN-GB"> _<b
 +
style=""><span style="color: rgb(255, 102, 0);"> </span></b>BB0<b
 +
style=""><span style="color: rgb(255, 102, 0);">57<span style="">&nbsp; </span></span></b>(Luciferase)</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border: 1pt solid windowtext; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Step
 +
7</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Vector</span></b></p>
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">digestion:
 +
SpeI + PstI</span></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><b style=""><span style="font-size: 10pt;">Insert
 +
      </span></b></p>
 +
      <p class="MsoNormal" style="text-indent: -5.4pt;"><span
 +
style="font-size: 10pt;"><span style="">&nbsp; </span>digestion: </span><span
 +
style="font-size: 8.5pt;">XbaI + PstI</span><span
 +
style="font-size: 10pt;"></span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">&nbsp;</span></p>
 +
      </td>
 +
      <td rowspan="16"
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 180pt;"
 +
valign="top" width="240">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;">BBa-J52017_CMV</span></p>
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt;">SP</span></b><span
 +
style="font-size: 10pt;">_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span></b></span><b style=""><span
 +
style="font-size: 10pt; color: rgb(153, 51, 0);" lang="EN-GB">scFv-anti-NIP</span></b><span
 +
style="font-size: 10pt;">_<b style="">GGGS-Li</b>_<b style=""><span
 +
style="color: rgb(153, 153, 153);">egfR-Tm</span>_<span
 +
style="color: rgb(255, 102, 0);">N</span></b>-</span><span
 +
style="font-size: 10pt;" lang="EN-GB">&#946;</span><span
 +
style="font-size: 10pt;">-Lactamase</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt; color: purple;" lang="EN-GB"> </span></b><b
 +
style=""><span style="font-size: 10pt;">SP</span></b><span
 +
style="font-size: 10pt;">_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span></b></span><b style=""><span
 +
style="font-size: 10pt; color: rgb(153, 51, 0);" lang="EN-GB">scFv-anti-NI</span></b><span
 +
style="font-size: 10pt;" lang="EN-GB"> </span><span
 +
style="font-size: 10pt;">_<b style="">GGGS-Li</b>_<b style=""><span
 +
style="color: rgb(153, 153, 153);"> egfR-Tm</span></b>_<b style=""><span
 +
style="color: rgb(255, 102, 0);">C</span></b>-</span><span
 +
style="font-size: 10pt;" lang="EN-GB">&#946;</span><span
 +
style="font-size: 10pt;">-Lactamase</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt;"> SP</span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(153, 51, 0);">scFv-anti-NIP</span></b>_<b style="">GGGS-Li</b>_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_<span style="color: blue;">SPLIT-Linker</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_</span><b style=""><span
 +
style="font-size: 10pt; color: rgb(255, 102, 0);">C</span></b><span
 +
style="font-size: 10pt;">-YFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt;"> SP</span></b><span
 +
style="font-size: 10pt;">_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span></b></span><b style=""><span
 +
style="font-size: 10pt; color: rgb(153, 51, 0);" lang="EN-GB">scFv-anti-NIP</span></b><span
 +
style="font-size: 10pt;">_<b style="">GGGS-Li</b>_<b style=""><span
 +
style="color: rgb(153, 153, 153);"> egfR-Tm</span>_<span
 +
style="color: rgb(255, 102, 0);">N</span></b>-YFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_</span><b
 +
style=""><span style="font-size: 10pt;"> SP</span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(153, 51, 0);">scFv-anti-NIP</span></b>_<b style="">GGGS-Li</b>_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_<span style="color: blue;">SPLIT-Linker</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_<b style=""><span
 +
style="color: rgb(255, 102, 0);">C</span></b>-CFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""> SP</b>_<b style=""><span style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(153, 51, 0);">scFv-anti-NIP</span></b>_<b style="">GGGS-Li</b>_<b
 +
style=""><span style="color: rgb(153, 153, 153);"> egfR-Tm</span></b>_<b
 +
style=""><span style="color: rgb(255, 102, 0);">N</span></b>-CFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""> SP</b>_<b style=""><span style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(153, 51, 0);">scFv-anti-NIP</span></b>_<b style="">GGGS-Li<span
 +
style="color: maroon;"> _</span><span
 +
style="color: rgb(153, 153, 153);"> egfR-Tm</span></b>_BB0<b style=""><span
 +
style="color: rgb(255, 102, 0);">58 </span></b>(Luciferase)</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""> SP</b>_<b style=""><span style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(153, 51, 0);">scFv-anti-NIP</span></b>_<b style="">GGGS-Li<span
 +
style="color: maroon;"> _</span><span
 +
style="color: rgb(153, 153, 153);"> egfR-Tm</span></b>_BB0<b style=""><span
 +
style="color: rgb(255, 102, 0);">57<span style="">&nbsp; </span></span></b>(Luciferase)</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: purple;"> </span>SP</b>_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span></b> _<b style="">GGGS-Li</b>_<b
 +
style=""><span style="color: rgb(153, 153, 153);"> egfR-Tm</span>_<span
 +
style="color: rgb(255, 102, 0);">N</span></b>-&#946;-Lactamase</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: purple;"> </span>SP</b>_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span></b> _<b style="">GGGS-Li</b>_<b
 +
style=""><span style="color: rgb(153, 153, 153);"> egfR-Tm</span>_<span
 +
style="color: rgb(255, 102, 0);">C</span></b>-&#946;-Lactamase</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: purple;"> </span></b></span><b style=""><span
 +
style="font-size: 10pt;">SP</span></b><span style="font-size: 10pt;"
 +
lang="EN-GB">_<b style=""><span style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span></b> _<b style="">GGGS-Li</b>_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_<span style="color: blue;">SPLIT-Linker</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_<b style=""><span
 +
style="color: rgb(255, 102, 0);"> C</span></b>-YFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: purple;"> </span></b></span><b style=""><span
 +
style="font-size: 10pt;">SP</span></b><span style="font-size: 10pt;"
 +
lang="EN-GB">_<b style=""><span style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span></b> _<b style="">GGGS-Li</b>_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_<span style="color: rgb(255, 102, 0);">N</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">-YFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: purple;"> </span></b></span><b style=""><span
 +
style="font-size: 10pt;">SP</span></b><span style="font-size: 10pt;"
 +
lang="EN-GB">_<b style=""><span style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span></b> _<b style="">GGGS-Li</b>_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><b style=""><span style="font-size: 10pt;"
 +
lang="EN-GB">_<span style="color: blue;">SPLIT-Linker</span></span></b><span
 +
style="font-size: 10pt;" lang="EN-GB">_<b style=""><span
 +
style="color: rgb(255, 102, 0);"> C</span></b>-CFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: purple;"> </span></b></span><b style=""><span
 +
style="font-size: 10pt;">SP</span></b><span style="font-size: 10pt;"
 +
lang="EN-GB">_<b style=""><span style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span></b> _<b style="">GGGS-Li</b>_</span><b
 +
style=""><span style="font-size: 10pt; color: rgb(153, 153, 153);">
 +
egfR-Tm</span></b><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: rgb(255, 102, 0);">N</span></b>-CFP</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: purple;"> </span>SP</b>_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span></b> _<b style="">GGGS-Li</b>_<b
 +
style=""><span style="color: maroon;">_</span><span
 +
style="color: rgb(153, 153, 153);"> egfR-Tm</span></b> _<b style=""><span
 +
style="color: rgb(255, 102, 0);"> </span></b>BB0<b style=""><span
 +
style="color: rgb(255, 102, 0);">58 </span></b>(Luciferase)</span></p>
 +
      </td>
 +
    </tr>
 +
    <tr style="">
 +
      <td
 +
style="border-style: none solid none none; border-color: -moz-use-text-color windowtext -moz-use-text-color -moz-use-text-color; border-width: medium 1pt medium medium; padding: 0cm 5.4pt; width: 41.4pt;"
 +
valign="top" width="55">
 +
      <p class="MsoNormal"><b style=""><span
 +
style="font-size: 10pt; color: purple;" lang="EN-GB">&nbsp;</span></b></p>
 +
      </td>
 +
      <td
 +
style="border-style: none solid solid none; border-color: -moz-use-text-color windowtext windowtext -moz-use-text-color; border-width: medium 1pt 1pt medium; padding: 0cm 5.4pt; width: 288pt;"
 +
valign="top" width="384">
 +
      <p class="MsoNormal"><span style="font-size: 10pt;" lang="EN-GB">_<b
 +
style=""><span style="color: purple;"> </span>SP</b>_<b style=""><span
 +
style="color: rgb(51, 102, 255);"> </span><span
 +
style="color: rgb(255, 102, 0);">Lipocalin</span></b> _<b style="">GGGS-Li</b>_<b
 +
style=""><span style="color: maroon;">_</span><span
 +
style="color: rgb(153, 153, 153);"> egfR-Tm</span></b> _<b style=""><span
 +
style="color: rgb(255, 102, 0);"> </span></b>BB0<b style=""><span
 +
style="color: rgb(255, 102, 0);">57<span style="">&nbsp; </span></span></b>(Luciferase)</span></p>
 +
      </td>
 +
    </tr>
 +
</table>
}}
}}

Revision as of 19:13, 27 October 2008


Freiburg2008 small header.gif



Home

The Team

Project Report

Parts

Modeling

Notebook

Safety

CoLABoration

Contents

Introduction:

This year´s main project is the attempt to create an "artificial receptor-system", featuring extra- and intracellular modules as well as suitable transmembrane regions. The intracellular domaine of our receptor-device is build by halves of split reporter-proteins that can reassemble and will then produce readable output, e. g. fluorescence. Each one of these protein-halves is connected to its extracellular domaine by a single-span transmembrane-helix. The extracellular or detecting domaine consists of a protein or peptide with the ability to bind a certain molecule.
Now, if a system with two matching receptors is presented these molecules in a strict, pairwise spatial arrangement, the receptor-devices are brought together, the split reporter-protein reassembles inside the cell and the output can be detected. We employ so-called "Origami-DNA" to create the exactly defined molecule-patterns that are needed to activate our receptors.

Material and Methods:

DNA-Origami

Results:

Discussion:

Literature:

Tabelle fuer Kathrin

Step 1

Vector

digestion: EcoRI + PstI

Insert

digestion: EcoRI + PstI

 

BBa-J52017

_CMV-promotor

Step 2

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _ SPLIT-Linker

 

C-YFP

 

C-CFP

Step 3

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _egfR-Tm

 

_  N-β-Lactamase

 

_  C-β-Lactamase

 

_ SPLIT-Linker_ C-YFP

 

_ N-YFP

 

_ SPLIT-Linker_ C-CFP

 

_ N-CFP

 

_ BB058 (Luciferase)

 

_ BB057  (Luciferase)

Step 4

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _SP

 

_scFv-anti-NIP

 

_ Lipocalin

Step 5

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _SP_ scFv-anti-NIP

and

pMA-BBFR-+SP_ Lipocalin

_GGGS-linker (produced by Klenow fill in)

Step 6

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _SP_ scFv-anti-NIP _ GGGS-Li

and

pMA-BBFR _ SP_ Lipocalin _

GGGS-Li

_ egfR-Tm _  N-β-Lactamase

 

_ egfR-Tm _  C-β-Lactamase

 

_ egfR-Tm _ SPLIT-Linker_ C-YFP

 

_ egfR-Tm _ N-YFP

 

_ egfR-Tm _ SPLIT-Linker_ C-CFP

 

_ egfR-Tm _ N-CFP

 

_ egfR-Tm _ BB058 (Luciferase)

 

_ egfR-Tm _ BB057  (Luciferase)

Step 7

Vector

digestion: SpeI + PstI

Insert

  digestion: XbaI + PstI

 

BBa-J52017_CMV

 

_SP_ scFv-anti-NIP_GGGS-Li_egfR-Tm_N-β-Lactamase

 

_ SP_ scFv-anti-NI _GGGS-Li_ egfR-Tm_C-β-Lactamase

 

_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_SPLIT-Linker_C-YFP

 

_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_N-YFP

 

_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_SPLIT-Linker_C-CFP

 

_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_N-CFP

 

_ SP_ scFv-anti-NIP_GGGS-Li _ egfR-Tm_BB058 (Luciferase)

 

_ SP_ scFv-anti-NIP_GGGS-Li _ egfR-Tm_BB057  (Luciferase)

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_N-β-Lactamase

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_C-β-Lactamase

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_SPLIT-Linker_ C-YFP

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_N-YFP

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_SPLIT-Linker_ C-CFP

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_N-CFP

 

_ SP_ Lipocalin _GGGS-Li__ egfR-Tm _ BB058 (Luciferase)

 

_ SP_ Lipocalin _GGGS-Li__ egfR-Tm _ BB057  (Luciferase)

Freiburg08 FT3.png

Retrieved from "http://2008.igem.org/Modular_Synthetic_Receptor_System"