Part:BBa_K4046600
tdTomato (red fluorescence gene)
tdTomato is a red fluorescent protein that was modified from previous fluorescence proteins in order to have enhanced fluorescence properties. When excited with yellow light, this protein fluoresces red light. The brightness of a sample should be correlated with the amount of protein present, which allows for dynamic tracking of the concentration of a compound of interest.
In order to test the expression of this gene, we created two different plasmids. One contained the tdTomato gene downstream of a CMV promoter, and the other one contained the tdTomato gene downstream of a hUBC promoter.
Figure 1: Gel images for tdTomato
These constructs were transfected into HEK cells, and plated cells were tested for fluorescence activity two days later. Overall, the imaging data shows that transfected cells demonstrate fluorescent activity as expected, validating the utilized sequence. Previous iGEM work has utilized this gene (BBa_J96029). However, our sequence is noted to differ from the one published in this part.
Figure 2: Microscopy images from hUBC-tdTomato construct
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 337
Illegal PstI site found at 1063 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 337
Illegal PstI site found at 1063 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 337
Illegal PstI site found at 1063 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 337
Illegal PstI site found at 1063 - 1000COMPATIBLE WITH RFC[1000]
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