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Gold Medal

Part:BBa_K2948005

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:

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.

Bronze Medal

Part:BBa_K2559003

We linked the gene of ATAPX1 to the existing promoter PrplJ, measured its enzyme activity in different states (different temperature, different illumination, different pH), and obtained correlation results between different factors and expressed enzyme protein activity.

Figure 1: This figure shows L-ascorbate peroxidase activity under different temperature, while ATAPX1 is control by PrplJ.

Figure 2: This figure shows L-ascorbate peroxidase activity under different illumination, while ATAPX1 is control by PrplJ.

Figure 3: This figure shows L-ascorbate peroxidase activity under different pH, while ATAPX1 is control by PrplJ.

Data which are obtained by our results, the above results indicate that we verify of the type of the promoter that PrplJ stability and reusability, especially in the experimental projects require different promoter can play a good role, such as the need to build contains multiple promoter and the need to use Gisbon assembly which is required to prevent the homologous recombination failure. When there is a must to use different promoter, PrplJ is a reliable choice.

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© 2019 Team:GZHS-United IGEM

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