Difference between revisions of "Part:BBa K3739023"

 
 
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Tevp can cut Ag43, leaving pelB as a signal peptide
 
Tevp can cut Ag43, leaving pelB as a signal peptide
  
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===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 04:58, 21 October 2021


pelB-Ag43-160N-Tevp

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 54
    Illegal NgoMIV site found at 892
    Illegal NgoMIV site found at 2116
    Illegal NgoMIV site found at 2626
    Illegal NgoMIV site found at 2647
    Illegal AgeI site found at 1642
    Illegal AgeI site found at 2386
    Illegal AgeI site found at 2800
    Illegal AgeI site found at 3042
  • 1000
    COMPATIBLE WITH RFC[1000]