Difference between revisions of "Part:BBa K3739058"
 
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<partinfo>BBa_K3739058 short</partinfo>
 
<partinfo>BBa_K3739058 short</partinfo>
  
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Aly01 is a signal peptide from the alginate lyase of  V.natriegens and GFP can show the secretion effciency of Aly01.
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Ag43 is a surface display protein and GFP can show the expression effciency of Ag43.
  
 
===Usage and Biology===
 
===Usage and Biology===
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Ag43 is a surface display protein that displays the protein molecule (GFP) it carries on the cell membrane of bacteria. pelB is an exopeptide that binds to Ag43, but is unable to do its job here, secreting the protein out of the bacteria.
 
Ag43 is a surface display protein that displays the protein molecule (GFP) it carries on the cell membrane of bacteria. pelB is an exopeptide that binds to Ag43, but is unable to do its job here, secreting the protein out of the bacteria.
  
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We used <partinfo>BBa_J23100</partinfo> to construct the expression system and obtained the composite <partinfo>BBa_K3739058</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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===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 05:01, 21 October 2021


J23100-B0030-pelB-Ag43-160N-his-GFP-B0010

Ag43 is a surface display protein and GFP can show the expression effciency of Ag43.

Usage and Biology

Ag43 is a surface display protein that displays the protein molecule (GFP) it carries on the cell membrane of bacteria. pelB is an exopeptide that binds to Ag43, but is unable to do its job here, secreting the protein out of the bacteria.

We used BBa_J23100 to construct the expression system and obtained the composite BBa_K3739058, 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.

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
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 118
    Illegal NgoMIV site found at 1076
    Illegal NgoMIV site found at 2300
    Illegal NgoMIV site found at 2810
    Illegal NgoMIV site found at 2831
    Illegal AgeI site found at 1826
    Illegal AgeI site found at 2570
    Illegal AgeI site found at 2984
    Illegal AgeI site found at 3226
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 206