Part:BBa_K2994001
Temperature Sensitive GFP Production Construct
Produces Temperature Sensitive GFP
Initial observation shows that gTEMP changes its intensity when temperature changes. Further quantitative testings are required to confirm these observations.
Usage and Biology
Figure 1: Gel for gTEMP. Lane 1 is the long ladder and Lanes 2-8 are K2994001. The white box shows the area where the bands are located in. The bands are in the correct size. The gel is poorly imaged due to the light source.
Figure 2: Test Data for gTEMP. Data were collected using a SpectroVis Plus from Vernier with the excitation wavelength set at 405nm and the peak emission from 410-415nm. If the construct were working, we would expect a linear increase in absorbance as temperature increases. These points are clearly not linear and not significantly different from each other (n=8). Bars represent average ± SEM.
Possible reasons for the lack of function: The recommended excitation wavelength is 360nm and our equipment cannot use that wavelength. No one in our local area has a spectrophotometer/fluorometer that can use the recommended wavelength. The codon usage was optimized for use in E. coli using the IDT website. This should not have changed the function, but It is possible that this did alter the function. We cannot use the sequence without optimization due to repeats that make the complexity score too high for IDT to synthesize.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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