TD1, Transdermal peptide
TD1 is a short synthetic peptide(ACSSSPSKHCG) identified by in vivo phage display, facilitated efficient transdermal protein delivery through intact skin. Studies suggested that the peptide creates a transient opening in the skin barrier to enable macromolecular material to reach systemic circulation.
Pgrac+RBS+SamyQ+TD1
+GFP
This is the main circuit of our project to allow high expression of target protein(antigen,adjuvant).GFP can be substituted by various proteins via a modular PCR or standard cut/ligation method.
PHT43
PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis.
HBsAg
HBsAg is the surface antigen of the hepatitis B virus (HBV). It indicates current hepatitis B infection.
Ag85b
The antigen 85 proteins (FbpA, FbpB, FbpC) are responsible for the high affinity of mycobacteria for fibronectin, a large adhesive glycoprotein, which facilitates the attachment of M.tuberculosis to murine alveolar macrophages (AMs).
Protective Antigen Domain 4
Protective antigen (PA) is the central component of the three-part protein toxin secreted by Bacillus anthracis, the organism responsible for anthrax.
LTB
This eltB gene encodes for the Heat-labile enterotox(LT) in certain virulent strains of E.coli.
Pctc+RBS+amilCP
Promoter ctc(BBa_K143010) is a sigma factor B-dependent promoter in B. subtilis. activated by endogenous sigma factor B under mild stress( nutrient stress response or physical stress response).
SdpA,SdpB,SpbC
Killing factor of Bacillus subtilis ,SpbC Induces the lysis of other B.subtilis cells that have not entered the sporulation pathway, providing a source of nutrients to support sporulation.
Pgrac+RBS+SDP
Spbc in SDP(sdpA,sdpB,spbC)(BBa_K1074009)operon is a killing factor of Bacillus subtilis.
PsdpRI+RBS+SDP
We construct this gene circuit as our kill switch to kill the engineered Bacillus subtilis for the safety purpose.Killing is mediated by the exported toxic protein SpbC.
Pgrac promoter
Prgac promoter(consisting of the groE promoter,the lacO operator and the gsiBSD sequence) allow induction by addition of ITPG.
promoter sdpR/I
This part is a sigma factor A-dependent promotor of the gene sdpR derived from the B.subtilis ,and it contains an operon for the signaling pathway of sdpc,a bacterial toxin of B.subtilis.
SamyQ
SamyQ is the signal peptide of the amyQ gene encoding an α-amylase in Bacillus subtilis WB800N. Fuse it with the recombinant proteins to obtain the secretion of recombinant proteins in Bacillus subtilis.
RBS for Pgrac
An efficient RBS for promoter Pgrac(BBa_K1074012) in Bacillus subtilis. We used it in all of our gene circuits containing promoter Pgrac.
RBS for Pctc
This is an efficient RBS for promoter Pctc(BBa_K143010) in Bacillus subtilis. It's also an universal RBS for promoters in Bacillus subtilis. We also used it with the promoter PsdpRI(BBa_K1074013) in our project.
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