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Team:GeorgiaTech/Biobricks
From 2010.igem.org
BioBricks also known as standard biological parts, are DNA sequences that when introduced into an organism, act as a component of a desired mechanism. For more information on BioBricks visit iGEM's BioBrick page here. |
The construct above is a fusion of the HybB cold shock promoter, the Ompa signal peptide, and AOXa. Aoxa is an alternative oxidase found in Sacred Lotus and shuffles electrons through an alternative path, thereby producing a thermogenesis event in Sacred Lotus. Our construct aims to produce an increase of heat above ambient temperature when cells expressing the construct are cold shocked (20c). The HybB promoter will express at lower temperatures (20 to 30c) but not at higher temperatures (37c). More information can be found at the construct page: |
The construct above containing mRFP (red fluorescent protein) at the c-terminus of AOX will allow for imaging in the bacterial periplasm. |
To quantify the activity of the HyBb promoter at different temperatures, we will express mRFP using HyBb in the construct above. Note that the final BioBricks will be in the pSB1C3 assembly plasmid. |
CONTROL: Picture shows nonlocalized mRFP expression throughout E Coli. They carry the biobrick sequence for mRFP expression (which is why they are red.) in psb1a3. They were grown overnight at 37C. |
EXPERIMENTAL: Picture shows localized periplasmic mRFP expression in E coli. These cells carry the coldshock promoter hybB, the targeting sequence for periplasmic expression ompA, and the red flourescent protein mRFP. This shows that we can direct the sequence to the periplasm. |
