Composite

Part:BBa_K2948005

Designed by: Zhang Xinyu   Group: iGEM19_GZHS-United   (2019-10-16)


Heme Oxygenase with GroE & RBS (E. coli Codon Optimised)

Hemeoxygenase with RBS (E. coli Codon Optimised), sequences obtained from BBa_K953000, and we turned the T7 promoter(Part BBa_I712074) into GroE promoter (BBa_K763001) This complex can then absorb red light (620-750 nm) to excite the bacteriophytochrome and result in a phenotypic change from blue to green. This can be reversed by far-red light (700-800 nm) or will revert from green to blue over time. As E. coli does not produce biliverdin, heme oxygenase must be coupled with a bacteriophytochrome to activate the oxidation of heme to produce biliverdin.

We changed the sequence of heme device in BBa K953000. We replace the T7 promoter with the promoter GroE. By taking photos and comparing the effects of different induction temperatures of plasmid pGroE-heme on the expression of target protein, we explored the expression induced by promoter GroE at different induction temperatures. The experimental results are as follows:

T--GZHS-United--Gold-Westernblot-GroE.png

Figure 1: It can be inferred from the figure that the change of T7 promoter to GroE promoter has a positive effect on protein expression in higher temperature.

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 312
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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