Difference between revisions of "Part:BBa K5301002"
Guanjiaxin (Talk | contribs) (→Characterization) |
Guanjiaxin (Talk | contribs) (→Characterization) |
||
| Line 13: | Line 13: | ||
===Characterization=== | ===Characterization=== | ||
| − | We used SDS-PAGE to test whether the protein containing SdyTag had been expressed successfully(Figure 1). The molecular weight of SnCSdT (containing SdyTag) is 42.1 kDa.And we purified the target protein with a molecular weight of approximately 43 kD (Lane 5 - Lane 8), which demonstrated that we had successfully expressed the protein containing SdyTag. | + | We used SDS-PAGE to test whether the protein containing SdyTag had been expressed successfully(Figure 1). The molecular weight of SnCSdT (containing SdyTag) is 42.1 kDa. And we purified the target protein with a molecular weight of approximately 43 kD (Lane 5 - Lane 8), which demonstrated that we had successfully expressed the protein containing SdyTag. |
<div class="center"><div class="thumb tnone"><div class="thumbinner" style="width:min-content;"><div style="zoom:0.5;overflow:hidden;"> | <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/sds-page-result-of-sncsdt-for-sdytag.png | https://static.igem.wiki/teams/5301/parts/sds-page-result-of-sncsdt-for-sdytag.png | ||
Revision as of 13:05, 30 September 2024
SdyTag can achieve covalent binding of proteins through Tag-Catcher interaction.
SdyTag is a fibronectin-binding protein. It can cooperate with SdyCatcher to achieve covalent binding of proteins through Tag-Catcher interaction. SdyTag and SdyCatcher can also be used alongside SpyTag-SpyCatcher partners, to demonstrate kinetically controlled directed protein ligation, domain specific protein circularization and macromolecular assembly [1].
Usage and Biology
We devised the fusion expression of SdyTag with spNW15 and SnoopCatcher (namely SnCSdT).We employed SdyTag and SdyCatcher to link two proteins - SCSdC-mCh[1-10] and SnCSdT.
Characterization
We used SDS-PAGE to test whether the protein containing SdyTag had been expressed successfully(Figure 1). The molecular weight of SnCSdT (containing SdyTag) is 42.1 kDa. And we purified the target protein with a molecular weight of approximately 43 kD (Lane 5 - Lane 8), which demonstrated that we had successfully expressed the protein containing SdyTag.
Figure 1.SDS-PAGE analysis of the extraction results of SnCSdT (containing SdyTag).The gel was run at 80 V for 10 minutes and then at 150 V for 20 minutes, followed by staining with Coomassie Brilliant Blue. The molecular weight of SnCSdT-M15-GS is 42.1 kDa.
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]
- ↑ Tan, L.L., S.S. Hoon and F.T. Wong, Kinetic Controlled Tag-Catcher Interactions for Directed Covalent Protein Assembly. PLOS ONE, 2016. 11(10): p. e0165074.