BACULOVIRUS EXPRESSION VECTOR
The invention concerns a baculovirus expression vector for recombinantly expressing an FMDV capsid precursor protein under control of a promoter, the expression vector comprising a nucleic acid sequence encoding the FMDV capsid precursor protein and the translational enhancers Syn2 land p10UTR. The invention further relates to a host cell comprising the baculovirus expression vector, a method of producing FMDV virus-like particles (VLPs), and a method of producing a vaccine.
1 . A baculovirus expression vector capable of recombinantly expressing a Foot and mouth disease virus (FMDV) capsid precursor protein under control of a promoter, the expression vector comprising:
(i) a nucleic acid sequence encoding the FMDV capsid precursor protein,
(ii) a translational enhancer Syn21 located within the 5′ untranslated region (UTR) of the nucleic acid sequence (i) encoding the FMDV capsid precursor protein, and
(iii) a translational enhancer P10UTR, located within the 3′UTR of the nucleic acid sequence (i) encoding the FMDV capsid precursor protein.
2 . The baculovirus expression vector according to claim 1 ,
wherein the translational enhancer Syn21 has a nucleic acid sequence corresponding to the nucleic acid sequence of SEQ ID NO. 1.
3 . The baculovirus expression vector according to claim 1 ,
wherein the translational enhancer P10UTR has a nucleic acid sequence corresponding to the nucleic acid sequence of SEQ ID NO. 2.
4 . The baculovirus expression vector according to claim 1 , wherein the FMDV is of the A serotype.
5 . The baculovirus expression vector according to claim 1 , wherein the FMDV is of the O serotype.
6 . The baculovirus expression vector according to claim 1 , wherein the capsid precursor protein comprises the capsid precursor P1.
7 . The baculovirus expression vector according to claim 1 , wherein the vector further comprises:
(iv) a nucleic acid sequence encoding a protease capable of cleaving the capsid precursor protein into one or more capsid proteins.
8 . The baculovirus expression vector according to claim 7 , wherein the capsid precursor protein comprises the capsid precursor P1 and the 2A peptide and the protease is 3C.
9 . A host cell comprising the baculovirus expression vector according to claim 1 .
10 . The host cell according to claim 9 , which is an insect cell.
11 . A method of producing FMDV capsid precursor protein, the method comprising the steps of:
(i) infecting a host cell with the baculovirus expression vector according to claim 1 , and
(ii) harvesting FMDV capsid precursor protein produced by the host cell.
12 . A method of producing FMDV virus-like particles (VLPs), the method comprising the steps of:
(i) infecting a host cell with the baculovirus expression vector according to claim 7 , and
(ii) harvesting FMDV VLPs produced by the host cell.
13 - 14 . (canceled)
15 . A method of producing a vaccine, which comprises the steps of:
(i) producing FMDV VLPs by the method according to claim 12 and
(ii) incorporating the FMDV VLPs into a vaccine by addition of a pharmaceutically acceptable carrier.
16 . A method of protecting a subject against an infection with FMDV, which comprises the step of expressing an FMDV capsid precursor protein from the baculovirus expression vector according to claim 7 in a host cell to produce a VLP, incorporating the VLP into a vaccine by addition of a pharmaceutically acceptable carrier and administering the vaccine to the subject.
17 . (canceled)