Difference between revisions of "Team:UNSW Australia/Results"
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<h2>Hypervesiculation Strain Library</h2> | <h2>Hypervesiculation Strain Library</h2> | ||
<p>Our first step was to generate some strains of <i>E. coli</i> that our research suggested would hypervesiculate. These are shown below:</p> | <p>Our first step was to generate some strains of <i>E. coli</i> that our research suggested would hypervesiculate. These are shown below:</p> | ||
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<img src="https://static.igem.org/mediawiki/2016/9/92/T--UNSW_Australia--StrainLibrary.png"> | <img src="https://static.igem.org/mediawiki/2016/9/92/T--UNSW_Australia--StrainLibrary.png"> | ||
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<h2>Assembly of g3p, tolR, PP-RFP and INPNC-GFP Expression Plasmids</h2> | <h2>Assembly of g3p, tolR, PP-RFP and INPNC-GFP Expression Plasmids</h2> | ||
<p>Our knock-in mutations were achieved by expressing g3p, TolR, PP-RFP, or INPNC-GFP in a T7-promoter driven vector (either pET or pRSF Duet). Final assembly of these plasmids was achieved using Gibson assembly.</p> | <p>Our knock-in mutations were achieved by expressing g3p, TolR, PP-RFP, or INPNC-GFP in a T7-promoter driven vector (either pET or pRSF Duet). Final assembly of these plasmids was achieved using Gibson assembly.</p> | ||
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<img src="https://static.igem.org/mediawiki/2016/d/d7/T--UNSW_Australia--CloningIntoDuet.png" style="vertical-align:middle" align="center"> | <img src="https://static.igem.org/mediawiki/2016/d/d7/T--UNSW_Australia--CloningIntoDuet.png" style="vertical-align:middle" align="center"> | ||
| − | <p>Figure 2A: Gel electrophoresis showing PP-RFP (lane 2, 1203bp) and INPNC-GFP (lane 3, 2133bp) inserted into the pET Duet expression plasmid. Lane 1 (554) is a negative control of pET with no insert.< | + | <p><i><b>Figure 2A:</b> Gel electrophoresis showing PP-RFP (lane 2, 1203bp) and INPNC-GFP (lane 3, 2133bp) inserted into the pET Duet expression plasmid. Lane 1 (554) is a negative control of pET with no insert. <b>Figure 2B</b>: Gel electrophoresis showing g3p inserted into the pRSF Duet expression plasmid (649bp). Lane 1 (551bp) is a negative control of pRSF with no insert. <b>Figure 2C</b>: Gel electrophoresis showing TolR (Lane 1, 687bp) inserted into the pET Duet expression plasmid. Lane 2 (554) is a negative control of pET with no insert.</i></p> |
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</div> | </div> | ||
</div> | </div> | ||
</section> | </section> | ||
Revision as of 00:57, 20 October 2016
Hypervesiculation Strain Library
Our first step was to generate some strains of E. coli that our research suggested would hypervesiculate. These are shown below:
Assembly of g3p, tolR, PP-RFP and INPNC-GFP Expression Plasmids
Our knock-in mutations were achieved by expressing g3p, TolR, PP-RFP, or INPNC-GFP in a T7-promoter driven vector (either pET or pRSF Duet). Final assembly of these plasmids was achieved using Gibson assembly.
Figure 2A: Gel electrophoresis showing PP-RFP (lane 2, 1203bp) and INPNC-GFP (lane 3, 2133bp) inserted into the pET Duet expression plasmid. Lane 1 (554) is a negative control of pET with no insert. Figure 2B: Gel electrophoresis showing g3p inserted into the pRSF Duet expression plasmid (649bp). Lane 1 (551bp) is a negative control of pRSF with no insert. Figure 2C: Gel electrophoresis showing TolR (Lane 1, 687bp) inserted into the pET Duet expression plasmid. Lane 2 (554) is a negative control of pET with no insert.
