Team:Hong Kong HKUST/results

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

Experiment 4: Cell Viability & Fatty Acid Quantification

Measuring cell viability at different fatty acid concentration & measure fatty acid uptake rate

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 theaceA 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 Pcmv 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.