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Team:Hong Kong HKUST/future
From 2013.igem.org
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| - | <strong>Construction of <i>aceB</i> constitutive expression construct | + | <strong>Construction of <i>aceB</i> constitutive expression construct.</strong> While all components of our intended <i>aceB</i> construct were obtained, the construct is yet to be completed. The full construct with EF-1alpha promoter, mitochondrial leader sequence, <i>aceB</i> gene, reporter protein and polyadenylation sequence are to be assembled in pSB1C3 backbone.<br><br> |
| - | <strong>Tests of promoter efficiency | + | <strong>Tests of promoter efficiency.</strong> GRP78 and FABP1 promoter efficiency can be measured by fluorescence intensity of green fluorescence protein at different fatty acid concentrations.<br><br> |
| - | <strong>Construction of inducible glyoxylate shunt construct | + | <strong>Construction of inducible glyoxylate shunt construct.</strong> The inducible promoter is to be fused with two functional genes, <i>aceA</i> and <i>aceB</i>.<br><br> |
| - | <strong>Transfection of construct for the inducible glyoxylate shunt | + | <strong>Transfection of construct for the inducible glyoxylate shunt.</strong> The construct developed to establish the inducible glyoxylate shunt is to be transfected into HepG2 cells to test its functionality. We plan to place the cell in media of containing different concentrations of fatty acids. The fatty acid uptake rate of our cells engineered for inducible glyoxylate shunt will be compared with that of those expressing constitutive glyoxylate shunt. |
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Revision as of 16:23, 27 September 2013
Future Work
Construction of aceB constitutive expression construct. While all components of our intended aceB construct were obtained, the construct is yet to be completed. The full construct with EF-1alpha promoter, mitochondrial leader sequence, aceB gene, reporter protein and polyadenylation sequence are to be assembled in pSB1C3 backbone.
Tests of promoter efficiency. GRP78 and FABP1 promoter efficiency can be measured by fluorescence intensity of green fluorescence protein at different fatty acid concentrations.
Construction of inducible glyoxylate shunt construct. The inducible promoter is to be fused with two functional genes, aceA and aceB.
Transfection of construct for the inducible glyoxylate shunt. The construct developed to establish the inducible glyoxylate shunt is to be transfected into HepG2 cells to test its functionality. We plan to place the cell in media of containing different concentrations of fatty acids. The fatty acid uptake rate of our cells engineered for inducible glyoxylate shunt will be compared with that of those expressing constitutive glyoxylate shunt.
