Difference between revisions of "Team:Macquarie Australia/Results"
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<li>Improved gBlock <i>hydG</i> which demonstrated a loss of functionality (2016) due to a point mutation.</li> | <li>Improved gBlock <i>hydG</i> which demonstrated a loss of functionality (2016) due to a point mutation.</li> | ||
<li>These gBlocks were inserted into one Biobrick (known as the Hydrogen Producing Gene Cluster) and transformed into <i>Escherichia coli</i> with a <i>lac</i> promoter and chloramphenicol resistance. </li> | <li>These gBlocks were inserted into one Biobrick (known as the Hydrogen Producing Gene Cluster) and transformed into <i>Escherichia coli</i> with a <i>lac</i> promoter and chloramphenicol resistance. </li> | ||
| − | <li>Once induced, we aimed to test the rate of hydrogen gas production in these cells.<li> | + | <li>Once induced, we aimed to test the rate of hydrogen gas production in these cells.</li> |
</ul> | </ul> | ||
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<ul> | <ul> | ||
| − | <li>Analyse, optimise and construct the necessary gBlocks.<li> | + | <li>Analyse, optimise and construct the necessary gBlocks.</li> |
| − | <li>Digest and ligate gblocks into Biobricks.<li> | + | <li>Digest and ligate gblocks into Biobricks.</li> |
| − | <li>Digest/Double digest in conjunction with sequencing to verify gBlocks.<li> | + | <li>Digest/Double digest in conjunction with sequencing to verify gBlocks.</li> |
| − | <li>Digest and ligate gBlocks together via standard assembly.<li> | + | <li>Digest and ligate gBlocks together via standard assembly.</li> |
| − | <li>Induce plasmid with IPTG for protein expression.<li> | + | <li>Induce plasmid with IPTG for protein expression.</li> |
| − | <li>Run cell lysate of <i>fer</i> on SDS-PAGE followed by Mass Spectrometry to analyse gel bands.<li> | + | <li>Run cell lysate of <i>fer</i> on SDS-PAGE followed by Mass Spectrometry to analyse gel bands.</li> |
| − | <li>Test hydrogen production using Clark electrode and gas volume measurement experiment.<li> | + | <li>Test hydrogen production using Clark electrode and gas volume measurement experiment.</li> |
</ul> | </ul> | ||
<br> | <br> | ||
Revision as of 10:39, 29 October 2017
Aim
- We designed and ordered the gBlocks for 4 genes, which encoded for proteins involved in hydrogen production (fer, hyd1, hydEF, hydG) from Chlamydomonas reinhardtii.
- Improved gBlock hydG which demonstrated a loss of functionality (2016) due to a point mutation.
- These gBlocks were inserted into one Biobrick (known as the Hydrogen Producing Gene Cluster) and transformed into Escherichia coli with a lac promoter and chloramphenicol resistance.
- Once induced, we aimed to test the rate of hydrogen gas production in these cells.
Experimental Design
- Analyse, optimise and construct the necessary gBlocks.
- Digest and ligate gblocks into Biobricks.
- Digest/Double digest in conjunction with sequencing to verify gBlocks.
- Digest and ligate gBlocks together via standard assembly.
- Induce plasmid with IPTG for protein expression.
- Run cell lysate of fer on SDS-PAGE followed by Mass Spectrometry to analyse gel bands.
- Test hydrogen production using Clark electrode and gas volume measurement experiment.
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