Composite

Part:BBa_K4147001

Designed by: Ana Belem García González   Group: iGEM22_Tec-Chihuahua   (2022-09-30)


PelB-(CBD)2-DrsB1: A dermaseptin antimicrobial peptide from Phyllomedusa bicolor

Among a wide range of AMPs, polycationic dermaseptin peptides are produced by the skin glands of the frog Phyllomedusa bicolor [1]. It supplies both antibacterial and antifungal protection to a broad range of plant pathogens but shows no toxic effects on plant and mammalian cells [1]. This 3.2 kDa peptide is fused to a tandem repeat of a chitin binding domain (CBD) from Cladosporium fulvumas an improvement of dermaseptin-b1 gene (AKA MsrA2)(BBa_K2577001) which binds to chitin fungal cell wall, thereby increasing the lytic activity of the catalytic domain. The peptide is fused with a PelB sequence (BBa_J32015) to send the protein to periplasmic space for an easier purification. The sequence also contains a 6xHis tag for purification with IMAC. The final protein has a molecular mass of 25.1 kDa.

Usage and Biology

This fusion of DrsB1 to CBD not only protect DrsB1 from host degradation but its affinity for fungal cell wall chitin remains the same. It has been shown showed that this recombinant peptide possessed strong antifungal activity in vitro, inhibiting the growth of mycelia as well as the germination of conidia and spores[1]. This part has been used in the generation of transgenic lines of Tobacco plants which were resistant to Alternaria, Phytophthora, Fusarium, Pythium fungi [2].

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 165
    Illegal AgeI site found at 444
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[1] Khademi, M., Varasteh-Shams, M., Nazarian-Firouzabadi, F., & Ismaili, A. (2020). New Recombinant Antimicrobial Peptides Confer Resistance to Fungal Pathogens in Tobacco Plants. Frontiers in plant science, 11, 1236. https://doi.org/10.3389/fpls.2020.01236

[2] Shams, M. V., Nazarian-Firouzabadi, F., Ismaili, A., & Shirzadian-Khorramabad, R. (2019). Production of a Recombinant Dermaseptin Peptide in Nicotiana tabacum Hairy Roots with Enhanced Antimicrobial Activity. Molecular Biotechnology. doi:10.1007/s12033-019-00153-x

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