Difference between revisions of "Part:BBa K4488017"
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freeuse Green Fluorescent Protein-B, a blue-shifted GFP that fluoresces under blue light. | freeuse Green Fluorescent Protein-B, a blue-shifted GFP that fluoresces under blue light. | ||
− | This part is an improvement of fuGFP (Part:BBa K3814004) which was originally designed to create a GFP variant not protected by patents and able to facilitate any kind of research. The original fuGFP absorbs less blue light compared to sfGFP. The decreased absorption of blue light is an issue for fuGFP because plate readers with special UV filters are required to detect the excitation wavelength, restricting its accessibility. Thus, using PCR with degenerate primers we introduced the S66T mutation in the fluorophore to create fuGFPb. The improved variant has a blue-shifted excitation peak (485 nm) (figure 1) and absorbs more blue light making it more similar to sfGFP. As a result, fuGFPb has the advantage of being more easily measured | + | This part is an improvement of fuGFP (Part:BBa K3814004) which was originally designed to create a GFP variant not protected by patents and able to facilitate any kind of research. The original fuGFP absorbs less blue light compared to sfGFP. The decreased absorption of blue light is an issue for fuGFP because plate readers with special UV filters are required to detect the excitation wavelength, restricting its accessibility. Thus, using PCR with degenerate primers we introduced the S66T mutation in the fluorophore to create fuGFPb. The improved variant has a blue-shifted excitation peak (485 nm) (figure 1) and absorbs more blue light making it more similar to sfGFP. As a result, fuGFPb has the advantage of being more easily measured as plate readers that do not extend into the UV range for excitation can be used. Furthermore, it has promising potential as a fluorescent tag. Additionally, we also created a fusion protein of fuGFPb with a cellulose-binding domain allowing it to be immobilised on cellulose (Part:BBa K4488020). |
===Usage and Biology=== | ===Usage and Biology=== |
Revision as of 01:06, 12 October 2022
fuGFPb
freeuse Green Fluorescent Protein-B, a blue-shifted GFP that fluoresces under blue light.
This part is an improvement of fuGFP (Part:BBa K3814004) which was originally designed to create a GFP variant not protected by patents and able to facilitate any kind of research. The original fuGFP absorbs less blue light compared to sfGFP. The decreased absorption of blue light is an issue for fuGFP because plate readers with special UV filters are required to detect the excitation wavelength, restricting its accessibility. Thus, using PCR with degenerate primers we introduced the S66T mutation in the fluorophore to create fuGFPb. The improved variant has a blue-shifted excitation peak (485 nm) (figure 1) and absorbs more blue light making it more similar to sfGFP. As a result, fuGFPb has the advantage of being more easily measured as plate readers that do not extend into the UV range for excitation can be used. Furthermore, it has promising potential as a fluorescent tag. Additionally, we also created a fusion protein of fuGFPb with a cellulose-binding domain allowing it to be immobilised on cellulose (Part:BBa K4488020).
Usage and Biology
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 151
- 1000COMPATIBLE WITH RFC[1000]