Difference between revisions of "Part:BBa K4147002"
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<partinfo>BBa_K4147002 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4147002 SequenceAndFeatures</partinfo>
  
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===Characterization of ===
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<center>[[File:Expression_construct_for_(CBD)2-DrsB1_sequence_7.png |300px]]</center>
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<center><b>Figure 1</b>. SnapGene®️ map of BioBrick BBa_K4147002.</center>
  
 
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Revision as of 21:14, 9 October 2022


Expression construct for (CBD)2-DrsB1: Dermaseptin AMP from Phyllomedusa bicolor

This part contains the linear construct BBa_K4147001 for (CBD)2-DrsB1 which already incorporates a PelB signal peptide on the N- terminal as well as a 6X-His tag at the end of the dermaseptin CDS (C- terminal) for later protein purification. This composite construct includes a LacI regulated promoter. Additionally, it contains an RBS under the part name BBa_B0032. Finally, it has a double terminator (BBa_B0015) to ensure correct termination. As a linear 1075 bp construct, it contains the RFC10 prefix and suffix, making the part compatible with other iGEM parts.


Usage and Biology

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 [2]. This peptide of molecular weight of 22.1 kDa contains a tandem repeat of a chitin binding domain (CBD) as 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.

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 392
    Illegal AgeI site found at 671
  • 1000
    COMPATIBLE WITH RFC[1000]

Characterization of

Expression construct for (CBD)2-DrsB1 sequence 7.png
Figure 1. SnapGene®️ map of BioBrick BBa_K4147002.


REFERENCES

[1] 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

[2] 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. doi:10.3389/fpls.2020.01236