Difference between revisions of "Part:BBa K3739113"

 
 
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<partinfo>BBa_K3739113 short</partinfo>
 
<partinfo>BBa_K3739113 short</partinfo>
  
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Tevp can cut Ag43, leaving pelB as a signal peptide.
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Tevp can cut Ag43, leaving pelB as a signal peptide
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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Ag43 is a surface display protein that displays protein molecule it carries on the cell membrane of bacteria. pelB is an exopeptide linked to Ag43. Tevp can shear Ag43 and release the exopeptide pelB to secrete the protein molecules out of bacteria.
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We used <partinfo>BBa_K525998</partinfo> to construct the expression system and obtained the composite <partinfo>BBa_K3739113</partinfo>, which is assembled on the expression vector pET-28(+) by standard assembly. The constructed plasmid was transformed into ''Vibrio natriegens'' , then the positive transformants were selected by kanamycin and confirmed by colony PCR and sequencing.
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===Reference===
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1. Ahan, R. E.; Kirpat, B. M.; Saltepe, B.; Seker, U. O. S., A Self-Actuated Cellular Protein Delivery Machine. ACS Synth Biol 2019, 8 (4), 686-696.
  
 
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Latest revision as of 05:04, 21 October 2021


K525998(T7-RBS)-pelB-Ag43-160N-Tevp-B0010

Tevp can cut Ag43, leaving pelB as a signal peptide

Usage and Biology

Ag43 is a surface display protein that displays protein molecule it carries on the cell membrane of bacteria. pelB is an exopeptide linked to Ag43. Tevp can shear Ag43 and release the exopeptide pelB to secrete the protein molecules out of bacteria.

We used BBa_K525998 to construct the expression system and obtained the composite BBa_K3739113, which is assembled on the expression vector pET-28(+) by standard assembly. The constructed plasmid was transformed into Vibrio natriegens , then the positive transformants were selected by kanamycin and confirmed by colony PCR and sequencing.

Reference

1. Ahan, R. E.; Kirpat, B. M.; Saltepe, B.; Seker, U. O. S., A Self-Actuated Cellular Protein Delivery Machine. ACS Synth Biol 2019, 8 (4), 686-696.

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 NgoMIV site found at 92
    Illegal NgoMIV site found at 930
    Illegal NgoMIV site found at 2154
    Illegal NgoMIV site found at 2664
    Illegal NgoMIV site found at 2685
    Illegal AgeI site found at 1680
    Illegal AgeI site found at 2424
    Illegal AgeI site found at 2838
    Illegal AgeI site found at 3080
  • 1000
    COMPATIBLE WITH RFC[1000]