Coding

Part:BBa_K5301002

Designed by: Jiaxin Guan   Group: iGEM24_BNU-China   (2024-09-21)
Revision as of 13:06, 30 September 2024 by Guanjiaxin (Talk | contribs)


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.

sds-page-result-of-sncsdt-for-sdytag.png

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===


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


  1. 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.
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