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Experiment 4: Cell Viability & Fatty Acid Quantification
Measuring cell viability at different fatty acid concentration & measure fatty acid uptake rate
Team:Hong Kong HKUST/results
From 2013.igem.org
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<li>Successfully constructed two parts</a> carrying mitochondrial leader sequence (MLS) compatible with RFC 10 and RFC 25 assembly standards respectively, (<a href="http://parts.igem.org/Part:BBa_K1119000">BBa_K1119000</a>, <a href="http://parts.igem.org/Part:BBa_K1119001">BBa_K1119001</a>).</li> | <li>Successfully constructed two parts</a> carrying mitochondrial leader sequence (MLS) compatible with RFC 10 and RFC 25 assembly standards respectively, (<a href="http://parts.igem.org/Part:BBa_K1119000">BBa_K1119000</a>, <a href="http://parts.igem.org/Part:BBa_K1119001">BBa_K1119001</a>).</li> | ||
| - | <li>Successfully constructed </a>parts containing the<i>aceA</i> and <i>aceB</i> elements of the glyoxylate shunt (<a href="http://parts.igem.org/Part:BBa_K1119002">BBa_K1119002</a>, <a href="http://parts.igem.org/Part:BBa_K1119003">BBa_K1119003</a>). </li> | + | <li>Successfully constructed </a>parts containing the <i>aceA</i> and <i>aceB</i> elements of the glyoxylate shunt (<a href="http://parts.igem.org/Part:BBa_K1119002">BBa_K1119002</a>, <a href="http://parts.igem.org/Part:BBa_K1119003">BBa_K1119003</a>). </li> |
<li>Successfully constructed a part containing the constitutive regulatory promoter of Human Elongation Factor 1-alpha (<a href="http://parts.igem.org/Part:BBa_K1119004">BBa_K1119004</a>).</li> | <li>Successfully constructed a part containing the constitutive regulatory promoter of Human Elongation Factor 1-alpha (<a href="http://parts.igem.org/Part:BBa_K1119004">BBa_K1119004</a>).</li> | ||
| - | <li>Successfully constructed a part containing the constitutive | + | <li>Successfully constructed a part containing the constitutive CMV promoter (<a href="http://parts.igem.org/Part:BBa_K1119006">BBa_K1119006</a>).</li> |
<li>Successfully characterized the constitutive promoter P<i>cmv</i> in HEK293FT cells (<a href="http://parts.igem.org/Part:BBa_K1119008">BBa_K1119008</a>).</li> | <li>Successfully characterized the constitutive promoter P<i>cmv</i> in HEK293FT cells (<a href="http://parts.igem.org/Part:BBa_K1119008">BBa_K1119008</a>).</li> | ||
<li>Successfully characterized the mitochondrial leader sequence (MLS) in HEK293FT cells (<a href="http://parts.igem.org/Part:BBa_K1119009">BBa_K1119009</a>).</li> | <li>Successfully characterized the mitochondrial leader sequence (MLS) in HEK293FT cells (<a href="http://parts.igem.org/Part:BBa_K1119009">BBa_K1119009</a>).</li> | ||
Revision as of 21:26, 27 September 2013
Result
Experiment 1: Glyoxylate Shunt
Introduce glyoxylate enzymes responsible for the shunt
Experiment 2: Protein Trafficking
Target ACE proteins into mitochondria
Experiment 3: Fatty Acid Sensing Mechanism
Introduce inducible system that allows tunable fatty acid uptake by sensing fatty acid concentration
Our project was first proposed in mid-March and after a series of improvement and elaboration, we started our wet lab work from mid-June to late-September. After 3 months of memorable diligent work, we are proud to claim that we have achieved the results below:
- Successfully constructed two parts carrying mitochondrial leader sequence (MLS) compatible with RFC 10 and RFC 25 assembly standards respectively, (BBa_K1119000, BBa_K1119001).
- Successfully constructed parts containing the aceA and aceB elements of the glyoxylate shunt (BBa_K1119002, BBa_K1119003).
- Successfully constructed a part containing the constitutive regulatory promoter of Human Elongation Factor 1-alpha (BBa_K1119004).
- Successfully constructed a part containing the constitutive CMV promoter (BBa_K1119006).
- Successfully characterized the constitutive promoter Pcmv in HEK293FT cells (BBa_K1119008).
- Successfully characterized the mitochondrial leader sequence (MLS) in HEK293FT cells (BBa_K1119009).
- Successfully tested HepG2 cell viability in different sodium palmitate concentrations.
