Composite

Part:BBa_K5066008

Designed by: Abner Tseng   Group: iGEM24_GEMS-Taiwan   (2024-09-21)

P20-Cyt2Ba

Description

The combination of Bt toxins and scorpion toxins is a promising strategy providing a potential layer of protection against insecticide resistance. The ribosome binding site(RBS) is a sequence of nucleotides before the start codon of an mRNA transcript and functions to recruit ribosomes for the translation of proteins. The His-tag, 6X His, is a commonly used purification tag that contains 6 consecutive histidine residues. The His-tag can be stained with His-tag antibodies after translation, labelling the target recombinant protein. It is typically placed on either the N or C terminus of a protein.

Use in Biology

p20 is a chaperone that aids in the proteins production of the Bt toxins. Cyt2Ba is a potent larvicide that works in the digestive tract of Aedes mosquitoes. The Cyt protein’s reactivity to alkaline conditions in Aedes mosquito larvae’s midgut causes it to solubilize by increasing membrane permeability, and undergo proteolytic cleavage. This activates the protein, leading to its binding to unsaturated phospholipids present in the cell membrane of the larvae’s epithelial cells lining the alimentary canal. Such a disruption in an epithelial cell’s membranes changes its membrane permeability, hindering normal cell processes, and ultimately, cell lysis. Once this takes place on a large scale, larvae midgut tissues lose their function and cause symptoms in the Aedes mosquito larvae including starvation, electrolyte imbalance and eventually death.[1][2][3]

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 264
    Illegal EcoRI site found at 1039
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 264
    Illegal EcoRI site found at 1039
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 264
    Illegal EcoRI site found at 1039
    Illegal BglII site found at 777
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 264
    Illegal EcoRI site found at 1039
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 264
    Illegal EcoRI site found at 1039
  • 1000
    COMPATIBLE WITH RFC[1000]

Reference

[1] Bravo, A., Likitvivatanavong, S., Gill, S. S., & Soberón, M. (2011). Bacillus thuringiensis: A story of a successful bioinsecticide. Insect Biochemistry and Molecular Biology, 41(7), 423–431. https://doi.org/10.1016/j.ibmb.2011.02.006

[2] Wu, J., Wei, L., He, J., Fu, K., Li, X., Jia, L., Wang, R., & Zhang, W. (2021). Characterization of a novel Bacillus thuringiensis toxin active against Aedes aegypti larvae. Acta Tropica, 223, 106088. https://doi.org/10.1016/j.actatropica.2021.106088

[3] Gu, J.-B., Dong, Y.-Q., Peng, H.-J., & Chen, X.-G. (2010). A Recombinant AeDNA Containing the Insect-Specific Toxin, BmK IT1, Displayed an Increasing Pathogenicity on Aedes albopictus. American Journal of Tropical Medicine and Hygiene, 83(3), 614–623. https://doi.org/10.4269/ajtmh.2010.10-0074

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