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Part:BBa_K4274005:Design

Designed by: Jiawen Chen   Group: iGEM22_KEYSTONE   (2022-09-30)


PmanP


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]


Design Notes

The manP promoter (PmanP) is situated in the upstream and same orientation of the mannose operon of Bacillus subtilis. It is reported that the presence of mannose could increase the beta-galactosidase activity, then result in a 4 to 7 fold increase in expression of lacZ[1]. Besides,this promoter is not active in E. coli, but is activated by manR in Bacillus subtilis, which constitutes the mannose operon of B. subtilis.Based on it, the pJOE8999 plasmid system, which allows genome editing of B. subtilis, was designed to carry cas9 gene under the control of the B. subtilis mannose-inducible promoter PmanP[2]. In this experiment, we are using a 0.2% concentration of mannose to induce cas9 expression of vector pJOE8999 to enable CRISPR-Cas9-based genome editing on B. subtilis strain 168.


Source

Bacillus subtilis

References