Composite

Part:BBa_K5520005:Design

Designed by: Ruyi Shi   Group: iGEM24_Ulink-SZ   (2024-09-29)


pT7-LacO-His-CsnB


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 844
    Illegal NotI site found at 804
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 813
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 153
    Illegal AgeI site found at 636
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We suggest that the second step of the reaction catalyzes the generation of chitosan from chitosan to chitooligosaccharides during the generation of chitosan from chitin. We utilized the chitosanase CsnB to degrade chitosan in order to obtain chitooligosaccharides with high added value, which can somewhat alter the reaction equilibrium of the precursor reaction and thus improve the utilization of the reaction substrate. We cloned the CsnB gene into the pET28a vector to obtain the PET-28a-CsnB recombinant plasmid, which was transformed into E.coli BL21 (DE3) for overexpression of chitosanase CsnB.


Source

Escherichia coli and Bacillus pumilus

References