Part:BBa_K2083011
Human Beta Globin With a Premature Termination Codon Fused to GFP Reporter
Coding region for human beta globin gene with GFP reporter fused to the end. The beta globin has it's first two exons fused together. There is a premature stop codon (PTC) before the intron. When being translated, the ribosome recognizes the exon junction that is formed when the intron is spliced out and sends the RNA for degradation. Therefore there should be little to no fluorescence. Overall, in experiments there was decreased production of GFP compared to BBa_K2083012.
Coding region for human beta globin with GFP reporter fused to the end. GFP fused to beta globin with a premature termination codon (PTC) can be found at BBa K2083011.
Figure 1: Wild Type Globin Fluorescence
Figure 2: PTC Globin Fluorescence
The validation of the globin reporters followed the same procedures as the ATG/ACG reporters. The wild type PTC globin reporter showed a much lower level of expression compared to the ATG reporter (BBa_K2083009) (14.77% of positive control).
Figure 3: Efficiency of WT/PTC β-globin reporters compared to positive control. Error bars represent standard error between normalized results from 3 biological replicates of the experiment.
For how the values were calculated see this: https://static.igem.org/mediawiki/2016/c/c7/T--WPI_Worcester--Imaging_Analysis.pdf
For the fluorescence values see here:
https://static.igem.org/mediawiki/2016/4/45/T--WPI_Worcester--globin_Trial1.pdf: Trial 1
https://static.igem.org/mediawiki/2016/8/89/T--WPI_Worcester--globin_Trial2.pdf: Trial 2
https://static.igem.org/mediawiki/2016/3/3d/T--WPI_Worcester--globin_Trial3.pdf: Trial 3
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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