Composite
Mut

Part:BBa_K4575013

Designed by: Sarah Hutzl, Sumohit Sharma   Group: iGEM23_Stuttgart   (2023-10-06)


Expression cassette for Mutanase expression in Pichia pastoris

DNA fragment which was designed for the expression of Mutanase from Penicillium rubens in the Pichia pastoris expression system. Mutanase hydrolizes alpha 1,3 glycosidic bonds of glucan polymers. Thus, it can attack the exopolysaccharides in a biofilm.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2440
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1224
    Illegal AgeI site found at 2161
    Illegal AgeI site found at 2594
  • 1000
    COMPATIBLE WITH RFC[1000]


Employment in the DentoZym project

The goal was to use BBa_K4575013 for expression and secretion of mutanase in Pichia pastoris. Overhangs for Gibson assembly were generated on BBa_K4575013 (Figure 1).


mut-for-registry.jpg

Figure 1: Mutanase expression cassette with overhangs for Gibson assembly. The gel was stained with MIDORIGreen advance DNA stain (NIPPON Genetics).


But Gibson assembly with two other fragments failed.


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