Difference between revisions of "Part:BBa K3606084"

 
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<partinfo>BBa_K3606084 short</partinfo>
 
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<p>To test the secretion efficiency of our 5 secretion peptides fused CaAP(Calcium Absorption Peptide) guided by 5 secretion peptides, we put them under the regulation of the iPTG inducible promoter, Ptac, and expressed them in E.coli. BL21 strain respectively. Thus, quantified characterization of these secretion peptides fused with CaAP is expected to be achieved through the different quantity of the target protein, namely CaAP, in the bacterial lysate and the culture medium.</p>
  
 
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<span class='h3bb'>Sequence and Features</span>
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<h4>Sequence and Features</h4>
 
<partinfo>BBa_K3606084 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3606084 SequenceAndFeatures</partinfo>
  
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===Characterization===
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<p>We have successfully cloned NSP4/PhoA/OmpA guided CaAP(Calcium Absorption Peptide) under the regulation of the iPTG inducible promoter, Ptac, and expressed it in E.coli.BL21. As shown in the figure below,PhoA can guide CaAP to secrete into the extracellular space.</p>
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[[File:T--Fudan--denoted_gel2_signalpeptides.png|none|400px|thumb|Figure1.NSP4/PhoA/OmpA-CaAP expression with and without iPTG induction in the bacterial lysate.]]
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[[File:T--Fudan--denoted_gel3_signalpeptides.png|none|400px|thumb|Figure2.NSP4/PhoA/OmpA-CaAP expression expression with and without iPTG induction in the culture medium concentrated by TCA.]]
  
 
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Revision as of 02:01, 28 October 2020


PlacIq_lacI_Ptac driven PhoA-CaAP

To test the secretion efficiency of our 5 secretion peptides fused CaAP(Calcium Absorption Peptide) guided by 5 secretion peptides, we put them under the regulation of the iPTG inducible promoter, Ptac, and expressed them in E.coli. BL21 strain respectively. Thus, quantified characterization of these secretion peptides fused with CaAP is expected to be achieved through the different quantity of the target protein, namely CaAP, in the bacterial lysate and the culture medium.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1233
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1414
  • 1000
    COMPATIBLE WITH RFC[1000]

Characterization

We have successfully cloned NSP4/PhoA/OmpA guided CaAP(Calcium Absorption Peptide) under the regulation of the iPTG inducible promoter, Ptac, and expressed it in E.coli.BL21. As shown in the figure below,PhoA can guide CaAP to secrete into the extracellular space.

Figure1.NSP4/PhoA/OmpA-CaAP expression with and without iPTG induction in the bacterial lysate.
Figure2.NSP4/PhoA/OmpA-CaAP expression expression with and without iPTG induction in the culture medium concentrated by TCA.