Coding

Part:BBa_K3381007

Designed by: Jeriann Hsiao   Group: iGEM20_Waterloo   (2020-10-21)
Revision as of 03:14, 22 October 2020 by Jlhsiao (Talk | contribs)

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CBM2a + Flexible Linker

3D model of the flexible linker (highlighted in green) between Mst-CopC, a copper-binding protein, and CBM2a (BBa_K863101)

To facilitate the use of our desired metal-binding proteins within a column, we sought to immobilize our proteins on cellulose. We first located the cellulose binding module 2a (CBM2a) in the iGEM parts registry under BBa_K863101. While the part should be functional without modification, we decided to insert a linker between our functional metal-binding protein and the CBM. We thought it may improve the efficiency of our separations by increasing the rate at which the column could equilibrate as the binding site would be less likely to be hidden against the cellulose surface. In our literature review, a paper described a naturally occurring flexible linker between a CBM2 domain and the catalytic domain in a lytic polysaccharide monooxygenase found in Streptomyces coelicolor A3 (Courtade et al., 2018). By including this linker, part BBa_K863101 would be functionally improved for applications where the CBM is used to immobilise the fused protein on cellulose. The effect of the presence of the linker could be tested by measuring the binding constants of both the metal-binding domain and the cellulose-binding domain as described on our Measurement page.


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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Categories
Parameters
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