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Team:Hong Kong HKUST/Project/module1
From 2013.igem.org
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Modules
- FA Quantification & Cell Viability
- FA Sensing Mechanism
- Protein Trafficking
- Glyoxylate Shunt
Fatty Acid Quantification and Cell Viability
Cell Line
For our project we made use of two mammalian cell lines: HepG2 human hepatoma cells, and HEK293FT human embryonic kidney cells. HepG2 cells were used for characterizing inducible promoters and the glyoxylate systems. To take advantage of their higher transfection efficiency, the characterizations of mitochondria leader sequence and constitutive promoter were conducted using HEK293FT cells.Cell Culture
HepG2 and HEK293FT cells were grown in DMEM supplemented with 10% heat-inactivated FBS, 50 μg/mL penicillin and 50 μg/mL streptomycin. The cells were incubated at 37℃ in a humidified atmosphere containing 5%CO2. Cells were transfected in petri dishes and multi-well plates with Lipofectamine 2000 (Invitrogen; Carlsbard, CA) transfection reagent according to the manufacturer’s protocols. GFP signals were observed under fluorescent microscope or under confocal microscope, if necessary.
