Composite

Part:BBa_K3819012

Designed by: Zhou Ying   Group: iGEM21_UESTC-China   (2021-09-28)


Self-design Scaffoldin protein

This is a scaffoldin protein designed by ourselves. It has two Ccscohesion, one CBM3, one Ctcohesion, and one Type II dockerin.

Finally, we successfully expressed this recombinant plasmid, and the Scaffoldin protein has been used in the following-up experiments of cellulosome assembly. And the expression results are as follows.

Figure 1. The Dot-Blot results of recombined GS115(pScaf) fermentation broth. The sample was gained after fermentation in YPD for 96h. Scaffoldin(Myc-Tag) was tested by adding 2ul fermentation supernatant on NC membrane and exposing 60s.


Figure 2. The SDS-PAGE analysis of recombined GS115(pScaf). Line1, 2, 3, 4, 5, 6 are concentrated GS115(pEGL7) supernate, GS115(pEGL7) supernate, GS115(pEGl7 ) lysate, concentrated GS115 supernate, GS115 supernate, and GS115 lysate respectively. Scaffoldin is 98.5KDa in theory but 130-180KDa in practice because of glycosylation.


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]


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