Intermediate

Part:BBa_S04059

Designed by: Doug Tischer   Group: Guests   (2008-08-17)


katG (LAA).B0015

Construction intermediate

Characterization by team Mingdao 2020

ldhp-AQP-RBS-SpxB-Tr-tpxp-KatG-Tr/pSB1C3 (BBa_K3376012)
Catalase converts H2O2 to O2 and H2O preventing cells from oxidative stress. When E. coli Nissle strain (EcN) was expressing aquaporin (AQP) and pyruvate oxidase (SpxB) to produce H2O2, the cell growth was inhibited remarkably compared to a vector-only control. By adding of catalase (KatG), the growth rate became normal and have similar response to glucose concentrations with the control.
T--Mingdao--ww5.png


The lysates of the transformed E. coli Nissle carrying the indicated gene were subjected to SDS-PAGE and Coomassie blue staining. When overexpression under the promoter of ldhp, KatG (80kDa) and AQP (17kDa) have sharp band on the PAGE. However, the predicted 65-kDa of SpxB was not detected, possibly because of the oxidative stress induced by the production of H2O2. In the presence of KatG, SpxB was shown up along with AQP, though expressed at a week level.
T--Mingdao--ww10.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 228
    Illegal BglII site found at 1163
    Illegal BglII site found at 1290
    Illegal BamHI site found at 1080
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None