Difference between revisions of "Part:BBa K3788011"
 
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BBa_K3788011 is coding for pelB, sfGFP and AidaA-I (D>N). pelB is a leader sequence, it allows the protein to join the periplasm via the SEC system (unfolded proteins). Aida-I, on the other hand, is an autotransporter from the T5SS; it allow the addressing to the extracellular medium, the mutation of the 846 – 847 amino acid permit to no have protein cleavage, and attach this one to the extracellular membrane. In this contruction sfGFP permit to understand the mechanism, this protein can at the end be replaced by an other protein. sfGFP is supposed to be address to the preriplasm thanks to pelB leader and the T5SS system will address sfGFP to the extracellular membrane without cleavage. Sequence was optimised for E. coli K12.
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NOTOC
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<partinfo>BBa_K3788011 short</partinfo>
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<p>BBa_K3788011 is coding for AidaA-I (D>N). It is an autotransporter from the T5SS; it allow the addressing to the extracellular medium, with a mutation that permit to have no protein cleavage, and keep it the passenger protein attached to the extracellular membrane.</p>
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<p>The sequence is coding for the beta-barrel sequence that forms a pore in the external membrane and a linker that allows a passenger protein to go through the beta barrel to join the extracellular medium.</p>
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<p>Wild-Type Aida-I has an autolytic activity that allows the passenger protein cleavage from the rest of the complex so it is released in the extracellular medium, but we introduced D:33N substitution the the cleavage site is not recognized anymore and the passenger protein remains attached to the cell.</p>
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<!-- Add more about the biology of this part here
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3788011 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K3788011 parameters</partinfo>
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Latest revision as of 15:52, 21 October 2021

NOTOC Aida-I (D33:N) T5SS


BBa_K3788011 is coding for AidaA-I (D>N). It is an autotransporter from the T5SS; it allow the addressing to the extracellular medium, with a mutation that permit to have no protein cleavage, and keep it the passenger protein attached to the extracellular membrane.

The sequence is coding for the beta-barrel sequence that forms a pore in the external membrane and a linker that allows a passenger protein to go through the beta barrel to join the extracellular medium.

Wild-Type Aida-I has an autolytic activity that allows the passenger protein cleavage from the rest of the complex so it is released in the extracellular medium, but we introduced D:33N substitution the the cleavage site is not recognized anymore and the passenger protein remains attached to the cell.


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 1084
  • 1000
    COMPATIBLE WITH RFC[1000]