Snow mountain virus genome sequence, virus-like particles and methods of use
View Patent ↗Snow Mountain Virus (SMV) belongs to the Norovirus genus of the Caliciviridae family. SMV is a genogroup II (GII) reference strain of human enteric caliciviruses associated with epidemic gastroenteritis. The positive sense RNA genome sequence of SMV was determined to be 7,537 nucleotides in length excluding the 3′ polyadenylated tract. The genome is organized into three open reading frames. Pairwise sequence alignments showed SMV ORF1 is highly conserved with other GII noroviruses, and most closely related to GII strains Melksham and Hawaii viruses. Comparative sequence analyses showed the SMV is a recombinant norovirus. VP1/NP2 proteins assembled into virus-like particles (VLPs) when expressed in insect cells by a recombinant baculovirus. Characterization of one clone that expressed VP1 but failed to assemble into VLPs, identified histidine residue 91 as important for particle assembly.
1. A composition comprising a virus-like particle comprising Snow Mountain virus VP1 protein and Snow Mountain virus VP2 protein.
2. The composition according to claim 1 , wherein said VP2 protein is greater than 98% identical to SEQ ID NO: 28.
3. The composition according to claim 1 , wherein said VP2 protein is SEQ ID NO: 28.
4. The composition according to claim 1 , wherein said VP2 protein is encoded by a nucleic acid that is greater than 97% identical to SEQ ID NO: 31.
5. The composition according to claim 1 , wherein said VP2 protein is encoded by a nucleic acid that encodes for an amino acid sequence that is greater than 98% identical to SEQ ID NO: 28.
6. The composition according to claim 1 , wherein said VP1 protein is greater than 98% identical to SEQ ID NO: 27.
7. The composition according to claim 1 , wherein said VP1 protein is SEQ ID NO:27.
8. The composition according to claim 1 , wherein said VP1 protein is encoded by a nucleic acid that is greater than 94% identical to SEQ ID NO: 30.
9. The composition according to claim 1 , wherein said VP1 protein is encoded by a nucleic acid identical to SEQ ID NO: 30.
10. The composition according to claim 1 , wherein said VP1 is encoded by a nucleic acid that encodes an amino acid sequence that is greater than 98% identical to SEQ ID NO: 27.
11. The composition according to claim 1 , wherein said VP1 protein is identical to SEQ ID NO: 27 and said VP2 protein is identical to SEQ ID NO: 28.
12. The composition according to claim 1 , wherein said composition further comprises and adjuvant and an amount of said virus-like particle suitable for inducing an immune response to a subject.
13. The composition according to claim 12 , wherein said immune response is an antibody response.
14. The composition according to claim 12 , wherein said composition further comprises a carrier.
15. A method of making a virus-like particle comprising co-expressing Snow Mountain virus VP1 protein and Snow Mountain virus VP2 protein in a recombinant nucleic acid expression system under conditions suitable for assembly of the expressed VP1 and VP2 proteins into a virus-like particle.
16. The method according to claim 15 , wherein said expression system comprises a eukaryotic cell.
17. The method according to claim 16 , wherein said eukaryotic cell is an insect cell.
18. The method according to claim 15 , wherein said system is a baculovirus expression system.
19. The method according to claim 15 , wherein said VP2 protein is greater than 98% identical to SEQ ID NO: 28.
20. The method according to claim 15 , wherein said VP2 protein is SEQ ID NO: 28.
21. The method according to claim 15 , wherein said VP2 protein is expressed from a nucleic acid that is greater than 97% identical to SEQ ID NO: 31.
22. The method according to claim 15 , wherein said VP2 protein is expressed from a nucleic acid that encodes for an amino acid sequence that is greater than 98% identical to SEQ ID NO: 28.
23. The method according to claim 15 , wherein said VP1 protein is greater than 98% identical to SEQ ID NO: 27.
24. The method according to claim 15 , wherein said VP1 protein is SEQ ID NO: 27.
25. The method according to claim 15 , wherein said VP1 protein is encoded by a nucleic acid that is greater than 94% identical to SEQ ID NO: 30.
26. The method according to claim 15 , wherein said VP1 protein is encoded by a nucleic acid identical to SEQ ID NO: 30.
27. The method according to claim 15 , wherein said VP1 protein is encoded by a nucleic acid that encodes an amino acid sequence that is greater than 98% identical to SEQ ID NO: 27.
28. A method of making an antibody comprising administering composition of claim 1 to an animal.
29. The method according to claim 17 , wherein said insect cell is an Sf9 cell.