Difference between revisions of "Part:BBa K3018000"

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[[Image:T--Tongji_China--indigo_powder.jpg|left|400px|thumb|Indigo powder extracted from culture medium]]
 
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[[Image:T--Tongji_China--tjindigo.jpg|left|400px|thumb|Tubes containing blue culture medium, color comes from IPTG induction. Tubes organized in the shape of TJ]]
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[[Image:T--Tongji_China--tjindigo.jpg|left|400px|thumb|Tubes containing blue culture medium, induced by IPTG. Tubes organized in the shape of TJ]]
 
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Revision as of 22:59, 21 October 2019


Bacterial flavin-containing monooxydase, codon-optimized

This part encodes a bacterial flavin-containing monooxygenase(bFMO), which can catalyze the oxidation of indole. We had the codon-optimized for E. coli. You can use this part to produce indigo through indoxyl and leucoindigo in E. coli. Indigo is a hydrophobic blue dye; the expression of functional FMO can be estimated by the color of culture solution. Clones on LB plates should be blue when it works.

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]


Tongji_China 2019 Characterization


All characterizations are performed in plasmid pET-28a(+), strain E. coli BL21(DE3).
Function Validation To validate the function of this part in E. coli BL21(DE3), we transformed pET-28a(+)-bFMO into E. coli BL21(DE3). Bacte and induced with IPTG (final concentration: 0.5mM) . Culture without induction was used as control.

Dark blue clones on LB plate with IPTG


Validation of function. Tube on the left was induced with IPTG, on the right was not induced as control. This figure shows a significant difference in color between induced group and control group, indicates the production of indigo. The result supports that this part can work in BL21(DE3).


Indigo powder extracted from culture medium


Tubes containing blue culture medium, induced by IPTG. Tubes organized in the shape of TJ