Part:BBa_K3606084
PlacIq_lacI_Ptac driven PhoA-CaAP
PlacIq_lacI_Ptac driven PhoA-CaAP is a part of our signal-peptide guided CaAP secretion system. <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.
The 5 signal peptides candidates for CaAP secretion are listed below.
Signal Peptide | Number for Registry |
---|---|
NSP4 | BBa_K3606042 |
OmpA | BBa_K3606043 |
DsbA | BBa_K3606030 |
PelB | BBa_K208004 |
PhoA | BBa_K808028 |
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1233
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1414
- 1000COMPATIBLE 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.
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