Difference between revisions of "Part:BBa K4727004"

 
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was based on the fundamental work of DA Marvin16, which suggested that mutants lacking the N2 domain can infect bacteria without pilus with a 100-fold higher efficiency16, as N2 masks the interaction of N1 with the co-receptor TolA. Building on this, the idea was to design a gene with a deleted N2 domain, drawing from the findings of Nilsson et al.10, who precisely delineated the amino acid regions corresponding to individual domains. Therefore, an alternative gene was designed for tropism that could ideally enable E. coli infection in the absence of a pilus, thus extending the tropism. This gene, named "Gp3_cut," was obtained by in silico deletion of the region between residues 87 and 256, corresponding to the N2 domain10. However, considering this protein's ability to interact with the TolA protein, specifically the TolAIII domain9,10, it was hypothesized that this same modified gene could also interact with homologous TolA proteins. Through bioinformatic analysis, using the BLASTp tool, a surface protein of Klebsiella pneumoniae, called "cell envelope integrity protein TolA" (protein ID: MBS4159696.1), was identified, sharing 100% similarity with the TolAIII domain of E. coli (residues 305-42113). This suggests that such modification could potentially allow the engineered bacteriophage's entry into K. pneumoniae as well.
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The design of this part was based on the fundamental work of DA Marvin[1], which suggested that mutants lacking the N2 domain og Gp3 protein of M13 bacteriphage can infect bacteria without pilus with a 100-fold higher efficiency[1], as N2 masks the interaction of N1 with the co-receptor TolA. Building on this, the idea was to design a gene with a deleted N2 domain, drawing from the findings of Nilsson et al.[2], who precisely delineated the amino acid regions corresponding to individual domains. Therefore, an alternative gene was designed for tropism that could ideally enable <i>E. coli</i> infection in the absence of a pilus, thus extending the tropism. This gene, named was obtained by in silico deletion of the region between residues 87 and 256, corresponding to the N2 domain[2].
  
 
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<span class='h3bb'>Sequence and Features</span>
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== Sequence and Features ==
 
<partinfo>BBa_K4727004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4727004 SequenceAndFeatures</partinfo>
  
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== References ==
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[1] Marvin, D. Filamentous phage structure, infection and assembly. Curr. Opin. Struct. Biol. 8, 150–158 (1998).
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[2] Nilsson, N., Malmborg, A.-C. & Borrebaeck, C. A. K. The Phage Infection Process: a Functional Role for the Distal Linker Region of Bacteriophage Protein 3. J. Virol. 74, 4229–4235 (2000)
  
 
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Latest revision as of 20:45, 24 July 2023


M13 Gene3 N2 deletion

The design of this part was based on the fundamental work of DA Marvin[1], which suggested that mutants lacking the N2 domain og Gp3 protein of M13 bacteriphage can infect bacteria without pilus with a 100-fold higher efficiency[1], as N2 masks the interaction of N1 with the co-receptor TolA. Building on this, the idea was to design a gene with a deleted N2 domain, drawing from the findings of Nilsson et al.[2], who precisely delineated the amino acid regions corresponding to individual domains. Therefore, an alternative gene was designed for tropism that could ideally enable E. coli infection in the absence of a pilus, thus extending the tropism. This gene, named was obtained by in silico deletion of the region between residues 87 and 256, corresponding to the N2 domain[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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 205

References

[1] Marvin, D. Filamentous phage structure, infection and assembly. Curr. Opin. Struct. Biol. 8, 150–158 (1998). [2] Nilsson, N., Malmborg, A.-C. & Borrebaeck, C. A. K. The Phage Infection Process: a Functional Role for the Distal Linker Region of Bacteriophage Protein 3. J. Virol. 74, 4229–4235 (2000)