Difference between revisions of "Part:BBa K4015002"

 
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<partinfo>BBa_K4015002 short</partinfo>
  
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DsbA is a soluble periplasmic protein. It belongs to the thioredoxin family and acts as a type of signal peptide which serve as the primary source of disulfide bonds to secreted proteins. The protein catalyzes the oxidation of the two cysteine sulfhydryls and form a covalent S-S bond connecting amino acid, then donates to the secreted protein, catalyzing their oxidation and their folding. DsbA’s size is about 21-kDa, primarily active in type II secretion peptide.
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===Usage and Biology===
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In our project, we pick three signal peptides to help secrete our target proteins, which are HlyA、GeneIII and DsbA .
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We used p47 vector containing GFP to compare the effectiveness of these signal peptides. p47-GFP-HlyA displayed a lower level of fluorescence comparing to p47-GeneIII-GFP, suggesting HlyA is more effective in secretion since more GFP has entered the medium. p47-DsbA-GFP showed no fluorescence, meaning protein expression was inhibited or the folding of the protein has failed. 
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<div>https://2021.igem.org/wiki/images/f/f2/T--KEYSTONE--GFP.png<div>
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 Fig. Result of GFP secretion experiment 
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4015002 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K4015002 parameters</partinfo>
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Revision as of 11:30, 20 October 2021

DsbA

DsbA is a soluble periplasmic protein. It belongs to the thioredoxin family and acts as a type of signal peptide which serve as the primary source of disulfide bonds to secreted proteins. The protein catalyzes the oxidation of the two cysteine sulfhydryls and form a covalent S-S bond connecting amino acid, then donates to the secreted protein, catalyzing their oxidation and their folding. DsbA’s size is about 21-kDa, primarily active in type II secretion peptide.

Usage and Biology

In our project, we pick three signal peptides to help secrete our target proteins, which are HlyA、GeneIII and DsbA . We used p47 vector containing GFP to compare the effectiveness of these signal peptides. p47-GFP-HlyA displayed a lower level of fluorescence comparing to p47-GeneIII-GFP, suggesting HlyA is more effective in secretion since more GFP has entered the medium. p47-DsbA-GFP showed no fluorescence, meaning protein expression was inhibited or the folding of the protein has failed. 

T--KEYSTONE--GFP.png

 Fig. Result of GFP secretion experiment 


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
    COMPATIBLE WITH RFC[1000]