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.
Claims23
A Bacillus exosporium molecule delivery (BEMD) system comprising a recombinant Bacillus cereus family member expressing a fusion construct, wherein the fusion construct comprises at least one molecule of interest (MOI) and:
- (a) a targeting sequence comprising amino acids 1-28 of SEQ ID NO:
- 10;
- (b) a targeting sequence comprising amino acids 13-28 of SEQ ID NO:
- 10;
- (c) a targeting sequence comprising SEQ ID NO:
- 10;
- or
- (d) a targeting sequence having at least 50% identity with amino acids 13-28 of SEQ ID NO:
- 10, wherein the identity with amino acids 18-28 of SEQ ID NO:
- 10 is at least 63%;
- and wherein expression of the fusion construct results in display of the MOI on the exosporium of the recombinant Bacillus cereus family member.
A method of making a delivery system comprising:
- a) constructing a fusion construct, wherein the fusion construct comprises at least one MOI and:
- (i) a targeting sequence comprising amino acids 1-28 of SEQ ID NO:
- 10;
- (ii) a targeting sequence comprising amino acids 13-28 of SEQ ID NO:
- 10;
- (iii) a targeting sequence comprising SEQ ID NO:
- 10;
- or
- (iv) a targeting sequence having at least 50% identity with amino acids 13-28 of SEQ ID NO:
- 10, wherein the identity with amino acids 18-28 of SEQ ID NO:
- 10 is at least 63%;
- b) cloning the fusion construct into a shuttle plasmid;
- and
- c) electroporating the shuttle plasmid containing the fusion construct into a Bacillus cereus family member, wherein expression of the fusion construct results in display of the MOI on the exosporium of the Bacillus cereus family member.
Dependencies are inferred from the claim text.