Bacillus based delivery system and methods of use
AssigneeThe Curators of the University of Missouri
InventorsGeorge C. Stewart, Brian Matthew Thompson
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Abstract
Herein a Bacillus exosporium antigen delivery (BEAD) system that provides a means to introduce recombinant proteins or small molecules into the exosporium of members of the B. cereus family of bacteria, i.e. B. anthracis, B. cereus, and B. thuringiensis, is disclosed. The system results in the surface display of recombinant proteins or small molecules such that they can stimulate an immune response. In addition, methods of making and using the system are described.
Claims20
A Bacillus exosporium molecule delivery (BEMD) system comprising a recombinant Bacillus bacterium expressing a fusion construct, wherein the fusion construct is expressed under the control of a sporulation-phase specific promoter and wherein the fusion construct comprises at least one molecule of interest (MOI) and:
- (a) a targeting sequence comprising an amino acid sequence having at least 43% identity with amino acids 20-35 of SEQ ID NO:
- 7, wherein the identity with amino acids 25-35 is at least 54%;
- (b) a targeting sequence comprising amino acids 1-35 of SEQ ID NO:
- 7;
- (c) a targeting sequence comprising amino acids 20-35 of SEQ ID NO:
- 7;
- (d) a targeting sequence comprising SEQ ID NO:
- 13;
- (e) a targeting sequence comprising SEQ ID NO:
- 7;
- (f) a targeting sequence comprising amino acids 1-27 of SEQ ID NO:
- 8;
- (g) a targeting sequence comprising amino acids 12-27 of SEQ ID NO:
- 8;
- (h) a targeting sequence comprising SEQ ID NO:
- 8;
- (i) a targeting sequence comprising an amino acid sequence having at least 43% identity with amino acids 12-27 of SEQ ID NO:
- 8;
- wherein the identity with amino acids 17-27 is at least 63%;
- (j) a targeting sequence comprising amino acids 1-38 of SEQ ID NO:
- 9;
- (k) a targeting sequence comprising amino acids 23-38 of SEQ ID NO:
- 9;
- (l) a targeting sequence comprising SEQ ID NO:
- 9;
- (m) a targeting sequence comprising an amino acid sequence having at least 43% identity with amino acids 23-38 of SEQ ID NO:
- 9, wherein the identity with amino acids 28-38 is at least 54%;
- (n) a targeting sequence comprising amino acids 1-28 of SEQ ID NO:
- 10;
- (o) a targeting sequence comprising amino acids 13-28 of SEQ ID NO:
- 10;
- (p) a targeting sequence comprising SEQ ID NO:
- 10;
- or
- (q) a targeting sequence comprising an amino acid sequence having at least 43% identity with amino acids 13-28 of SEQ ID NO:
- 10, wherein the identity with amino acids 18-28 is at least 54%;
- and wherein expression of the fusion construct results in display of the MOI on the exosporium of the Bacillus bacterium.
A Bacillus exosporium molecule delivery (BEMD) system comprising a recombinant Bacillus bacterium expressing a fusion construct, wherein the fusion construct comprises at least one molecule of interest (MOI) and a targeting sequence comprising a methionine residue directly covalently bound to a targeting sequence comprising an amino acid sequence comprising 16 amino acids having at least 43% identity with amino acids 20-35 of SEQ ID NO:
- 7, wherein the identity with amino acids 25-35 is at least 54%.
Dependencies are inferred from the claim text.