Team:TU Delft-Leiden/Project/Life science/curli/cloning/GoldenGate

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Revision as of 18:44, 12 October 2014

Curli Module - Cloning - Golden Gate Assembly

3 pieces of DNA had to be cloned into pSB1C3 to construct our BioBricks BBa_K1316013 and BBa_K1316014 . Both BioBricks encode a rhamnose promoter and the coding sequences of csgB and csgA or csgA-HIS. To work efficiently, we used the Golden Gate Assembly to construct our BioBricks. In this way, we were able to restrict and ligate all the DNA fragments at once (see figure 1).

Figure 1. **Golden Gate Assembly of BBa_K1316013 and BBa_K1316014 .** The BioBricks both encode a rhamnose promoter (red rectangle) and the coding sequences of *csgB* and *csgA* or *csgA-HIS*, indicated with grey arrows. The beam beneath the arrows visualizes the regions and relative sizes of the 3 DNA pieces that were ligated into pSB1C3.

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 <h2> Curli Module - Cloning - Golden Gate Assembly</h2>
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-To construct the BioBricks <a href="http://parts.igem.org/Part:BBa_K1316013">BBa_K1316013 </a> and <a href="http://parts.igem.org/Part:BBa_K1316014">BBa_K1316014 </a>, both encoding a rhamnose promoter and the coding sequences of <i>csgB</i> and <i>csgA</i> or <i>csgA-HIS</i>, 3 pieces of DNA had to be cloned into pSB1C3. To work efficiently, we used the LINK Golden Gate Assembly to construct our BioBricks, which made it possible to clone these BioBricks in one step only (see figure 1).
+pieces of DNA had to be cloned into pSB1C3 to construct our BioBricks <a href="http://parts.igem.org/Part:BBa_K1316013">BBa_K1316013 </a> and <a href="http://parts.igem.org/Part:BBa_K1316014">BBa_K1316014 </a>. Both BioBricks encode a rhamnose promoter and the coding sequences of <i>csgB</i> and <i>csgA</i> or <i>csgA-HIS</i>. To work efficiently, we used the Golden Gate Assembly to construct our BioBricks. In this way, we were able to restrict and ligate all the DNA fragments at once (see figure 1).
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