Difference between revisions of "Part:BBa K5301017"
| Line 3: | Line 3: | ||
<partinfo>BBa_K5301017 short</partinfo> | <partinfo>BBa_K5301017 short</partinfo> | ||
| − | sGFP1-10 tether is composed of mSA, a 3C linker, a | + | sGFP1-10 tether is composed of mSA, a 3C linker, a 6×His tag, and sGFP 1-10.We construct membrane protein dimers through the interaction between biotin and streptavidin, as well as the self-assembly mechanism of sGFP. |
===Usage and Biology=== | ===Usage and Biology=== | ||
| Line 13: | Line 13: | ||
We conducted colony PCR validation to assess the feasibility of the plasmid pathway, as shown in Figure 1, where the corresponding bands are very clear, indicating that the target fragments were successfully introduced into the plasmid and transformed into the bacterial strain. | We conducted colony PCR validation to assess the feasibility of the plasmid pathway, as shown in Figure 1, where the corresponding bands are very clear, indicating that the target fragments were successfully introduced into the plasmid and transformed into the bacterial strain. | ||
| − | <div class="center"><div class="thumb tnone"><div class="thumbinner" style="width:min-content;"><div style="zoom:0. | + | <div class="center"><div class="thumb tnone"><div class="thumbinner" style="width:min-content;"><div style="zoom:0.5;overflow:hidden;"> |
https://static.igem.wiki/teams/5301/parts/sgfp1-10-pcr.png | https://static.igem.wiki/teams/5301/parts/sgfp1-10-pcr.png | ||
</div><div class="thumbcaption"> | </div><div class="thumbcaption"> | ||
Revision as of 06:49, 1 October 2024
sGFP1-10 tether is composed of mSA, a 3C linker, a 6His tag, and sGFP 1-10.
sGFP1-10 tether is composed of mSA, a 3C linker, a 6×His tag, and sGFP 1-10.We construct membrane protein dimers through the interaction between biotin and streptavidin, as well as the self-assembly mechanism of sGFP.
Usage and Biology
mSA is used for binding to biotinylated proteins, and sGFP1-10 and sGFP11 self-assemble to form GFP, thereby creating a protein dimer that can report the outcome with fluorescence.
Characterization
We conducted colony PCR validation to assess the feasibility of the plasmid pathway, as shown in Figure 1, where the corresponding bands are very clear, indicating that the target fragments were successfully introduced into the plasmid and transformed into the bacterial strain.
Figure 1.PCR result of the sGFP1-10 tether.The base sequence length of the sGFP1-10 tether is 1071 base pairs (bp)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 82
Illegal AgeI site found at 142 - 1000COMPATIBLE WITH RFC[1000]