Difference between revisions of "Part:BBa K4882008"
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | Metarhizium anisopliae is an entomopathogenic fungus widely used as a biopesticide. To improve its safety, SuperNova is used as a part of a suicide switch for the engineered M. anisopliae. SuperNova is linked downstream of a hemolymph inducible promoter Pmcl1 (short | + | Metarhizium anisopliae is an entomopathogenic fungus widely used as a biopesticide. To improve its safety, SuperNova is used as a part of a suicide switch for the engineered M. anisopliae. SuperNova is linked downstream of a hemolymph inducible promoter Pmcl1 (short, [https://parts.igem.org/Part:BBa_K4882000 BBa BBa_K4882000]). When the fungi invade the insect body, Pmcl1(short) turns on, and they cumulate SuperNova protein. When the fungi grow out of the insect body and try to spread spores, the tissues are under sunlight and killed by ROS. |
===Characterization=== | ===Characterization=== |
Revision as of 13:44, 4 October 2023
SuperNova
SuperNova is a red fluorescent protein that produces reactive oxygen species (ROS) in the presence of green-yellow light (~579 nm) (Takemoto et al., 2013). Supernova is the monomeric version of KillerRed (BBa_K1184000) and should produce three times as much ROS as KillerRed can (Onukwufor et al., 2019).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 151
Illegal BsaI.rc site found at 442
Usage and Biology
Metarhizium anisopliae is an entomopathogenic fungus widely used as a biopesticide. To improve its safety, SuperNova is used as a part of a suicide switch for the engineered M. anisopliae. SuperNova is linked downstream of a hemolymph inducible promoter Pmcl1 (short, BBa BBa_K4882000). When the fungi invade the insect body, Pmcl1(short) turns on, and they cumulate SuperNova protein. When the fungi grow out of the insect body and try to spread spores, the tissues are under sunlight and killed by ROS.
Characterization
2023 Hangzhou-SDG Team characterized this part as part of a suicide switch
SuperNova can be a part of the suicide switch for M. anisopliae when being connected after the hemolymph inducible promoter Pmcl1/Pmcl1(short).
Larvae of Galleria mellonella were killed by engineered M. anisopliae. The corpses were placed under sunshine for days until spores formed outside the insect body. Spores were collected by vertexing and the concentrations of spore suspensions were checked under a microscope.
Results showed that suicide switches consisting of Supernova led to significantly fewer spores than WT did.
References
Onukwufor, J. O.; Trewin, A. J.; Baran, T. M.; Almast, A.; Foster, T. H.; Wojtovich, A. P. Quantification of Reactive Oxygen Species Production by the Red Fluorescent Proteins KillerRed, Supernova, and mCherry. Free Radical Biology and Medicine 2020, 147, 1–7. DOI:10.1016/j.freeradbiomed.2019.12.008.
Takemoto, K., Matsuda, T., Sakai, N. et al. SuperNova, a monomeric photosensitizing fluorescent protein for chromophore-assisted light inactivation. Sci Rep 3, 2629 (2013). https://doi.org/10.1038/srep02629