CN113402615A - 双位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原及其制备和应用 - Google Patents
双位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原及其制备和应用 Download PDFInfo
- Publication number
- CN113402615A CN113402615A CN202110553190.XA CN202110553190A CN113402615A CN 113402615 A CN113402615 A CN 113402615A CN 202110553190 A CN202110553190 A CN 202110553190A CN 113402615 A CN113402615 A CN 113402615A
- Authority
- CN
- China
- Prior art keywords
- cys
- gly
- ala
- thr
- plpe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000000427 antigen Substances 0.000 title claims abstract description 51
- 102000036639 antigens Human genes 0.000 title claims abstract description 51
- 108091007433 antigens Proteins 0.000 title claims abstract description 51
- 241000714203 Rabbit hemorrhagic disease virus Species 0.000 title claims abstract description 47
- 241000606860 Pasteurella Species 0.000 title claims abstract description 41
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 241000283973 Oryctolagus cuniculus Species 0.000 claims abstract description 48
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 31
- 229960005486 vaccine Drugs 0.000 claims abstract description 20
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 19
- 150000001413 amino acids Chemical class 0.000 claims abstract description 16
- 208000031169 hemorrhagic disease Diseases 0.000 claims abstract description 16
- 206010034107 Pasteurella infections Diseases 0.000 claims abstract description 14
- 201000005115 pasteurellosis Diseases 0.000 claims abstract description 14
- 230000003612 virological effect Effects 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 7
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 claims description 31
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 claims description 31
- 239000012634 fragment Substances 0.000 claims description 15
- 241000606856 Pasteurella multocida Species 0.000 claims description 11
- 229940051027 pasteurella multocida Drugs 0.000 claims description 11
- 210000004027 cell Anatomy 0.000 claims description 10
- 239000013612 plasmid Substances 0.000 claims description 9
- 229920001184 polypeptide Polymers 0.000 claims description 8
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 8
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 8
- 241000701447 unidentified baculovirus Species 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 5
- 239000013600 plasmid vector Substances 0.000 claims description 5
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 4
- 210000004899 c-terminal region Anatomy 0.000 claims description 4
- 238000010367 cloning Methods 0.000 claims description 4
- 239000002773 nucleotide Substances 0.000 claims description 4
- 125000003729 nucleotide group Chemical group 0.000 claims description 4
- 239000013604 expression vector Substances 0.000 claims description 2
- 239000013605 shuttle vector Substances 0.000 claims description 2
- 230000001131 transforming effect Effects 0.000 claims description 2
- 230000017105 transposition Effects 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 abstract description 15
- 238000002474 experimental method Methods 0.000 abstract description 8
- 239000000203 mixture Substances 0.000 abstract description 7
- 101710194807 Protective antigen Proteins 0.000 abstract description 5
- 230000008569 process Effects 0.000 abstract description 4
- 230000004224 protection Effects 0.000 abstract description 4
- 244000052769 pathogen Species 0.000 abstract 1
- 108010061238 threonyl-glycine Proteins 0.000 description 50
- 108010004073 cysteinylcysteine Proteins 0.000 description 41
- HYKFOHGZGLOCAY-ZLUOBGJFSA-N Cys-Cys-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O HYKFOHGZGLOCAY-ZLUOBGJFSA-N 0.000 description 27
- TVYMKYUSZSVOAG-ZLUOBGJFSA-N Cys-Ala-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O TVYMKYUSZSVOAG-ZLUOBGJFSA-N 0.000 description 25
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 25
- 108010016616 cysteinylglycine Proteins 0.000 description 24
- UWQJHXKARZWDIJ-ZLUOBGJFSA-N Ala-Ala-Cys Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CS)C(O)=O UWQJHXKARZWDIJ-ZLUOBGJFSA-N 0.000 description 21
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 21
- UQJNXZSSGQIPIQ-FBCQKBJTSA-N Gly-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)CN UQJNXZSSGQIPIQ-FBCQKBJTSA-N 0.000 description 20
- TVTZEOHWHUVYCG-KYNKHSRBSA-N Gly-Thr-Thr Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O TVTZEOHWHUVYCG-KYNKHSRBSA-N 0.000 description 20
- 108010047495 alanylglycine Proteins 0.000 description 20
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 19
- CVOZXIPULQQFNY-ZLUOBGJFSA-N Cys-Ala-Cys Chemical compound C[C@H](NC(=O)[C@@H](N)CS)C(=O)N[C@@H](CS)C(O)=O CVOZXIPULQQFNY-ZLUOBGJFSA-N 0.000 description 18
- ALNKNYKSZPSLBD-ZDLURKLDSA-N Cys-Thr-Gly Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O ALNKNYKSZPSLBD-ZDLURKLDSA-N 0.000 description 17
- IDOGEHIWMJMAHT-BYPYZUCNSA-N Gly-Gly-Cys Chemical compound NCC(=O)NCC(=O)N[C@@H](CS)C(O)=O IDOGEHIWMJMAHT-BYPYZUCNSA-N 0.000 description 16
- GVVKYKCOFMMTKZ-WHFBIAKZSA-N Gly-Cys-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CS)NC(=O)CN GVVKYKCOFMMTKZ-WHFBIAKZSA-N 0.000 description 15
- GQGAFTPXAPKSCF-WHFBIAKZSA-N Gly-Ala-Cys Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CS)C(=O)O GQGAFTPXAPKSCF-WHFBIAKZSA-N 0.000 description 14
- RCQRKPUXJAGEEC-ZLUOBGJFSA-N Ala-Cys-Cys Chemical compound C[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CS)C(O)=O RCQRKPUXJAGEEC-ZLUOBGJFSA-N 0.000 description 13
- AEJSNWMRPXAKCW-WHFBIAKZSA-N Cys-Ala-Gly Chemical compound SC[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O AEJSNWMRPXAKCW-WHFBIAKZSA-N 0.000 description 13
- UQJUGHFKNKGHFQ-VZFHVOOUSA-N Ala-Cys-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UQJUGHFKNKGHFQ-VZFHVOOUSA-N 0.000 description 12
- ZJBWJHQDOIMVLM-WHFBIAKZSA-N Cys-Cys-Gly Chemical compound SC[C@H](N)C(=O)N[C@@H](CS)C(=O)NCC(O)=O ZJBWJHQDOIMVLM-WHFBIAKZSA-N 0.000 description 12
- FTTZLFIEUQHLHH-BWBBJGPYSA-N Cys-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CS)N)O FTTZLFIEUQHLHH-BWBBJGPYSA-N 0.000 description 12
- JQFILXICXLDTRR-FBCQKBJTSA-N Gly-Thr-Gly Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)NCC(O)=O JQFILXICXLDTRR-FBCQKBJTSA-N 0.000 description 12
- KBBRNEDOYWMIJP-KYNKHSRBSA-N Thr-Gly-Thr Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O KBBRNEDOYWMIJP-KYNKHSRBSA-N 0.000 description 12
- DECCMEWNXSNSDO-ZLUOBGJFSA-N Ala-Cys-Ala Chemical compound C[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O DECCMEWNXSNSDO-ZLUOBGJFSA-N 0.000 description 11
- KRHRBKYBJXMYBB-WHFBIAKZSA-N Ala-Cys-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CS)C(=O)NCC(O)=O KRHRBKYBJXMYBB-WHFBIAKZSA-N 0.000 description 11
- OBVSBEYOMDWLRJ-BFHQHQDPSA-N Ala-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N OBVSBEYOMDWLRJ-BFHQHQDPSA-N 0.000 description 11
- SMYXEYRYCLIPIL-ZLUOBGJFSA-N Cys-Cys-Cys Chemical compound SC[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CS)C(O)=O SMYXEYRYCLIPIL-ZLUOBGJFSA-N 0.000 description 11
- XPNSAQMEAVSQRD-FBCQKBJTSA-N Thr-Gly-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)NCC(O)=O XPNSAQMEAVSQRD-FBCQKBJTSA-N 0.000 description 11
- KUFVXLQLDHJVOG-SHGPDSBTSA-N Ala-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](C)N)O KUFVXLQLDHJVOG-SHGPDSBTSA-N 0.000 description 10
- YFXFOZPXVFPBDH-VZFHVOOUSA-N Cys-Ala-Thr Chemical compound C[C@@H](O)[C@H](NC(=O)[C@H](C)NC(=O)[C@@H](N)CS)C(O)=O YFXFOZPXVFPBDH-VZFHVOOUSA-N 0.000 description 10
- CCQOOWAONKGYKQ-BYPYZUCNSA-N Gly-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)CN CCQOOWAONKGYKQ-BYPYZUCNSA-N 0.000 description 10
- 108010089804 glycyl-threonine Proteins 0.000 description 10
- 230000004083 survival effect Effects 0.000 description 10
- LJFNNUBZSZCZFN-WHFBIAKZSA-N Ala-Gly-Cys Chemical compound N[C@@H](C)C(=O)NCC(=O)N[C@@H](CS)C(=O)O LJFNNUBZSZCZFN-WHFBIAKZSA-N 0.000 description 9
- VNBNZUAPOYGRDB-ZDLURKLDSA-N Gly-Cys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)CN)O VNBNZUAPOYGRDB-ZDLURKLDSA-N 0.000 description 9
- DGOJNGCGEYOBKN-BWBBJGPYSA-N Thr-Cys-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CS)C(=O)O)N)O DGOJNGCGEYOBKN-BWBBJGPYSA-N 0.000 description 9
- LYGKYFKSZTUXGZ-ZDLURKLDSA-N Thr-Cys-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)NCC(O)=O LYGKYFKSZTUXGZ-ZDLURKLDSA-N 0.000 description 9
- COYHRQWNJDJCNA-NUJDXYNKSA-N Thr-Thr-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O COYHRQWNJDJCNA-NUJDXYNKSA-N 0.000 description 9
- 230000003053 immunization Effects 0.000 description 9
- 238000002649 immunization Methods 0.000 description 9
- WNHNMKOFKCHKKD-BFHQHQDPSA-N Ala-Thr-Gly Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O WNHNMKOFKCHKKD-BFHQHQDPSA-N 0.000 description 8
- URDUGPGPLNXXES-WHFBIAKZSA-N Cys-Gly-Cys Chemical compound SC[C@H](N)C(=O)NCC(=O)N[C@@H](CS)C(O)=O URDUGPGPLNXXES-WHFBIAKZSA-N 0.000 description 8
- NAPULYCVEVVFRB-HEIBUPTGSA-N Cys-Thr-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](N)CS NAPULYCVEVVFRB-HEIBUPTGSA-N 0.000 description 8
- 108010079364 N-glycylalanine Proteins 0.000 description 8
- WYKJENSCCRJLRC-ZDLURKLDSA-N Thr-Gly-Cys Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N[C@@H](CS)C(=O)O)N)O WYKJENSCCRJLRC-ZDLURKLDSA-N 0.000 description 8
- 108010084264 glycyl-glycyl-cysteine Proteins 0.000 description 8
- VGPWRRFOPXVGOH-BYPYZUCNSA-N Ala-Gly-Gly Chemical compound C[C@H](N)C(=O)NCC(=O)NCC(O)=O VGPWRRFOPXVGOH-BYPYZUCNSA-N 0.000 description 7
- DZLQXIFVQFTFJY-BYPYZUCNSA-N Cys-Gly-Gly Chemical compound SC[C@H](N)C(=O)NCC(=O)NCC(O)=O DZLQXIFVQFTFJY-BYPYZUCNSA-N 0.000 description 7
- NMROINAYXCACKF-WHFBIAKZSA-N Gly-Cys-Cys Chemical compound NCC(=O)N[C@@H](CS)C(=O)N[C@@H](CS)C(O)=O NMROINAYXCACKF-WHFBIAKZSA-N 0.000 description 7
- ZKJZBRHRWKLVSJ-ZDLURKLDSA-N Gly-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)CN)O ZKJZBRHRWKLVSJ-ZDLURKLDSA-N 0.000 description 7
- 241000699670 Mus sp. Species 0.000 description 7
- SLUWOCTZVGMURC-BFHQHQDPSA-N Thr-Gly-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O SLUWOCTZVGMURC-BFHQHQDPSA-N 0.000 description 7
- UQCNIMDPYICBTR-KYNKHSRBSA-N Thr-Thr-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O UQCNIMDPYICBTR-KYNKHSRBSA-N 0.000 description 7
- JBVSSSZFNTXJDX-YTLHQDLWSA-N Ala-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N JBVSSSZFNTXJDX-YTLHQDLWSA-N 0.000 description 6
- KOHBWQDSVCARMI-BWBBJGPYSA-N Cys-Cys-Thr Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KOHBWQDSVCARMI-BWBBJGPYSA-N 0.000 description 6
- NRVQLLDIJJEIIZ-VZFHVOOUSA-N Cys-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](CS)N)O NRVQLLDIJJEIIZ-VZFHVOOUSA-N 0.000 description 6
- FFJQHWKSGAWSTJ-BFHQHQDPSA-N Gly-Thr-Ala Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O FFJQHWKSGAWSTJ-BFHQHQDPSA-N 0.000 description 6
- YRNBANYVJJBGDI-VZFHVOOUSA-N Thr-Ala-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CS)C(=O)O)N)O YRNBANYVJJBGDI-VZFHVOOUSA-N 0.000 description 6
- QWMPARMKIDVBLV-VZFHVOOUSA-N Thr-Cys-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O QWMPARMKIDVBLV-VZFHVOOUSA-N 0.000 description 6
- NDZYTIMDOZMECO-SHGPDSBTSA-N Thr-Thr-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O NDZYTIMDOZMECO-SHGPDSBTSA-N 0.000 description 6
- TZQWJCGVCIJDMU-HEIBUPTGSA-N Thr-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)O)N)O TZQWJCGVCIJDMU-HEIBUPTGSA-N 0.000 description 6
- 241000700605 Viruses Species 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 description 5
- OEVCHROQUIVQFZ-YTLHQDLWSA-N Ala-Thr-Ala Chemical compound C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](C)C(O)=O OEVCHROQUIVQFZ-YTLHQDLWSA-N 0.000 description 5
- QKHWNPQNOHEFST-VZFHVOOUSA-N Ala-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](C)N)O QKHWNPQNOHEFST-VZFHVOOUSA-N 0.000 description 5
- CVLIHKBUPSFRQP-WHFBIAKZSA-N Cys-Gly-Ala Chemical compound [H]N[C@@H](CS)C(=O)NCC(=O)N[C@@H](C)C(O)=O CVLIHKBUPSFRQP-WHFBIAKZSA-N 0.000 description 5
- OXOQBEVULIBOSH-ZDLURKLDSA-N Cys-Gly-Thr Chemical compound [H]N[C@@H](CS)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O OXOQBEVULIBOSH-ZDLURKLDSA-N 0.000 description 5
- PYTZFYUXZZHOAD-WHFBIAKZSA-N Gly-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)CN PYTZFYUXZZHOAD-WHFBIAKZSA-N 0.000 description 5
- QSDKBRMVXSWAQE-BFHQHQDPSA-N Gly-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN QSDKBRMVXSWAQE-BFHQHQDPSA-N 0.000 description 5
- UZJDBCHMIQXLOQ-HEIBUPTGSA-N Thr-Cys-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O UZJDBCHMIQXLOQ-HEIBUPTGSA-N 0.000 description 5
- 230000035931 haemagglutination Effects 0.000 description 5
- 229940031551 inactivated vaccine Drugs 0.000 description 5
- 108010053725 prolylvaline Proteins 0.000 description 5
- RLMISHABBKUNFO-WHFBIAKZSA-N Ala-Ala-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O RLMISHABBKUNFO-WHFBIAKZSA-N 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 4
- CEXINUGNTZFNRY-BYPYZUCNSA-N Gly-Cys-Gly Chemical compound [NH3+]CC(=O)N[C@@H](CS)C(=O)NCC([O-])=O CEXINUGNTZFNRY-BYPYZUCNSA-N 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- IGROJMCBGRFRGI-YTLHQDLWSA-N Thr-Ala-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O IGROJMCBGRFRGI-YTLHQDLWSA-N 0.000 description 4
- TYVAWPFQYFPSBR-BFHQHQDPSA-N Thr-Ala-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)NCC(O)=O TYVAWPFQYFPSBR-BFHQHQDPSA-N 0.000 description 4
- 239000002671 adjuvant Substances 0.000 description 4
- 125000000539 amino acid group Chemical group 0.000 description 4
- 210000003719 b-lymphocyte Anatomy 0.000 description 4
- 238000013321 baculovirus-insect cell expression system Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 239000006069 physical mixture Substances 0.000 description 4
- 230000001681 protective effect Effects 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 229940031626 subunit vaccine Drugs 0.000 description 4
- CAJFZCICSVBOJK-SHGPDSBTSA-N Thr-Ala-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CAJFZCICSVBOJK-SHGPDSBTSA-N 0.000 description 3
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 230000005611 electricity Effects 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 238000010353 genetic engineering Methods 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 108010031424 isoleucyl-prolyl-proline Proteins 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- WOJJIRYPFAZEPF-YFKPBYRVSA-N 2-[[(2s)-2-[[2-[(2-azaniumylacetyl)amino]acetyl]amino]propanoyl]amino]acetate Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)CNC(=O)CN WOJJIRYPFAZEPF-YFKPBYRVSA-N 0.000 description 2
- SBGXWWCLHIOABR-UHFFFAOYSA-N Ala Ala Gly Ala Chemical compound CC(N)C(=O)NC(C)C(=O)NCC(=O)NC(C)C(O)=O SBGXWWCLHIOABR-UHFFFAOYSA-N 0.000 description 2
- MCKSLROAGSDNFC-ACZMJKKPSA-N Ala-Asp-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MCKSLROAGSDNFC-ACZMJKKPSA-N 0.000 description 2
- CZPAHAKGPDUIPJ-CIUDSAMLSA-N Ala-Gln-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(O)=O CZPAHAKGPDUIPJ-CIUDSAMLSA-N 0.000 description 2
- YMIYZAOBQDRCPP-UHFFFAOYSA-N Ala-Thr-Cys-Cys Chemical compound CC(N)C(=O)NC(C(O)C)C(=O)NC(CS)C(=O)NC(CS)C(O)=O YMIYZAOBQDRCPP-UHFFFAOYSA-N 0.000 description 2
- SZNGQSBRHFMZLT-IHRRRGAJSA-N Asn-Pro-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SZNGQSBRHFMZLT-IHRRRGAJSA-N 0.000 description 2
- 101800001383 Capsid protein VP60 Proteins 0.000 description 2
- FFVXLVGUJBCKRX-UKJIMTQDSA-N Gln-Ile-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CCC(=O)N)N FFVXLVGUJBCKRX-UKJIMTQDSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- CQGSYZCULZMEDE-SRVKXCTJSA-N Leu-Gln-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(O)=O CQGSYZCULZMEDE-SRVKXCTJSA-N 0.000 description 2
- CQGSYZCULZMEDE-UHFFFAOYSA-N Leu-Gln-Pro Natural products CC(C)CC(N)C(=O)NC(CCC(N)=O)C(=O)N1CCCC1C(O)=O CQGSYZCULZMEDE-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- ZIQQNOXKEFDPBE-BZSNNMDCSA-N Phe-Lys-His Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N ZIQQNOXKEFDPBE-BZSNNMDCSA-N 0.000 description 2
- FUMGHWDRRFCKEP-CIUDSAMLSA-N Ser-Leu-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O FUMGHWDRRFCKEP-CIUDSAMLSA-N 0.000 description 2
- HHJFMHQYEAAOBM-ZLUOBGJFSA-N Ser-Ser-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O HHJFMHQYEAAOBM-ZLUOBGJFSA-N 0.000 description 2
- PMTWIUBUQRGCSB-FXQIFTODSA-N Ser-Val-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O PMTWIUBUQRGCSB-FXQIFTODSA-N 0.000 description 2
- JZSLIZLZGWOJBJ-PMVMPFDFSA-N Trp-Phe-Lys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N JZSLIZLZGWOJBJ-PMVMPFDFSA-N 0.000 description 2
- VFOHXOLPLACADK-GVXVVHGQSA-N Val-Gln-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](C(C)C)N VFOHXOLPLACADK-GVXVVHGQSA-N 0.000 description 2
- JZWZACGUZVCQPS-RNJOBUHISA-N Val-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N JZWZACGUZVCQPS-RNJOBUHISA-N 0.000 description 2
- 230000004520 agglutination Effects 0.000 description 2
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 108010082286 glycyl-seryl-alanine Proteins 0.000 description 2
- 230000000521 hyperimmunizing effect Effects 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 230000003472 neutralizing effect Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- WXPZDDCNKXMOMC-AVGNSLFASA-N (2s)-1-[(2s)-2-[[(2s)-1-(2-aminoacetyl)pyrrolidine-2-carbonyl]amino]-5-(diaminomethylideneamino)pentanoyl]pyrrolidine-2-carboxylic acid Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@H](C(O)=O)CCC1 WXPZDDCNKXMOMC-AVGNSLFASA-N 0.000 description 1
- QMOQBVOBWVNSNO-UHFFFAOYSA-N 2-[[2-[[2-[(2-azaniumylacetyl)amino]acetyl]amino]acetyl]amino]acetate Chemical compound NCC(=O)NCC(=O)NCC(=O)NCC(O)=O QMOQBVOBWVNSNO-UHFFFAOYSA-N 0.000 description 1
- NJPMYXWVWQWCSR-ACZMJKKPSA-N Ala-Glu-Asn Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O NJPMYXWVWQWCSR-ACZMJKKPSA-N 0.000 description 1
- VWEWCZSUWOEEFM-WDSKDSINSA-N Ala-Gly-Ala-Gly Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(=O)NCC(O)=O VWEWCZSUWOEEFM-WDSKDSINSA-N 0.000 description 1
- NHLAEBFGWPXFGI-WHFBIAKZSA-N Ala-Gly-Asn Chemical compound C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)N)C(=O)O)N NHLAEBFGWPXFGI-WHFBIAKZSA-N 0.000 description 1
- SIGTYDNEPYEXGK-ZANVPECISA-N Ala-Gly-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)CNC(=O)[C@@H](N)C)C(O)=O)=CNC2=C1 SIGTYDNEPYEXGK-ZANVPECISA-N 0.000 description 1
- NINQYGGNRIBFSC-CIUDSAMLSA-N Ala-Lys-Ser Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CO)C(O)=O NINQYGGNRIBFSC-CIUDSAMLSA-N 0.000 description 1
- FEGOCLZUJUFCHP-CIUDSAMLSA-N Ala-Pro-Gln Chemical compound [H]N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O FEGOCLZUJUFCHP-CIUDSAMLSA-N 0.000 description 1
- XWFWAXPOLRTDFZ-FXQIFTODSA-N Ala-Pro-Ser Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O XWFWAXPOLRTDFZ-FXQIFTODSA-N 0.000 description 1
- RTZCUEHYUQZIDE-WHFBIAKZSA-N Ala-Ser-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RTZCUEHYUQZIDE-WHFBIAKZSA-N 0.000 description 1
- SYIFFFHSXBNPMC-UWJYBYFXSA-N Ala-Ser-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N SYIFFFHSXBNPMC-UWJYBYFXSA-N 0.000 description 1
- ARHJJAAWNWOACN-FXQIFTODSA-N Ala-Ser-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O ARHJJAAWNWOACN-FXQIFTODSA-N 0.000 description 1
- OMSKGWFGWCQFBD-KZVJFYERSA-N Ala-Val-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OMSKGWFGWCQFBD-KZVJFYERSA-N 0.000 description 1
- NABSCJGZKWSNHX-RCWTZXSCSA-N Arg-Arg-Thr Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H]([C@H](O)C)C(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N NABSCJGZKWSNHX-RCWTZXSCSA-N 0.000 description 1
- WVNFNPGXYADPPO-BQBZGAKWSA-N Arg-Gly-Ser Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O WVNFNPGXYADPPO-BQBZGAKWSA-N 0.000 description 1
- PCQXGEUALSFGIA-WDSOQIARSA-N Arg-His-Trp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O PCQXGEUALSFGIA-WDSOQIARSA-N 0.000 description 1
- HNJNAMGZQZPSRE-GUBZILKMSA-N Arg-Pro-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O HNJNAMGZQZPSRE-GUBZILKMSA-N 0.000 description 1
- ATABBWFGOHKROJ-GUBZILKMSA-N Arg-Pro-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O ATABBWFGOHKROJ-GUBZILKMSA-N 0.000 description 1
- KMFPQTITXUKJOV-DCAQKATOSA-N Arg-Ser-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O KMFPQTITXUKJOV-DCAQKATOSA-N 0.000 description 1
- HZPSDHRYYIORKR-WHFBIAKZSA-N Asn-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CC(N)=O HZPSDHRYYIORKR-WHFBIAKZSA-N 0.000 description 1
- XWGJDUSDTRPQRK-ZLUOBGJFSA-N Asn-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(N)=O XWGJDUSDTRPQRK-ZLUOBGJFSA-N 0.000 description 1
- IYVSIZAXNLOKFQ-BYULHYEWSA-N Asn-Asp-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O IYVSIZAXNLOKFQ-BYULHYEWSA-N 0.000 description 1
- JQSWHKKUZMTOIH-QWRGUYRKSA-N Asn-Gly-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)N)N JQSWHKKUZMTOIH-QWRGUYRKSA-N 0.000 description 1
- WIDVAWAQBRAKTI-YUMQZZPRSA-N Asn-Leu-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O WIDVAWAQBRAKTI-YUMQZZPRSA-N 0.000 description 1
- LZLCLRQMUQWUHJ-GUBZILKMSA-N Asn-Lys-Gln Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N LZLCLRQMUQWUHJ-GUBZILKMSA-N 0.000 description 1
- IDUUACUJKUXKKD-VEVYYDQMSA-N Asn-Pro-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O IDUUACUJKUXKKD-VEVYYDQMSA-N 0.000 description 1
- HNXWVVHIGTZTBO-LKXGYXEUSA-N Asn-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O HNXWVVHIGTZTBO-LKXGYXEUSA-N 0.000 description 1
- GOPFMQJUQDLUFW-LKXGYXEUSA-N Asn-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O GOPFMQJUQDLUFW-LKXGYXEUSA-N 0.000 description 1
- XPGVTUBABLRGHY-BIIVOSGPSA-N Asp-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)O)N XPGVTUBABLRGHY-BIIVOSGPSA-N 0.000 description 1
- CASGONAXMZPHCK-FXQIFTODSA-N Asp-Asn-Arg Chemical compound C(C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)O)N)CN=C(N)N CASGONAXMZPHCK-FXQIFTODSA-N 0.000 description 1
- UGIBTKGQVWFTGX-BIIVOSGPSA-N Asp-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)O)N)C(=O)O UGIBTKGQVWFTGX-BIIVOSGPSA-N 0.000 description 1
- SVFOIXMRMLROHO-SRVKXCTJSA-N Asp-Asp-Phe Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 SVFOIXMRMLROHO-SRVKXCTJSA-N 0.000 description 1
- KHGPWGKPYHPOIK-QWRGUYRKSA-N Asp-Gly-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O KHGPWGKPYHPOIK-QWRGUYRKSA-N 0.000 description 1
- KPNUCOPMVSGRCR-DCAQKATOSA-N Asp-His-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O KPNUCOPMVSGRCR-DCAQKATOSA-N 0.000 description 1
- DRCOAZZDQRCGGP-GHCJXIJMSA-N Asp-Ser-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O DRCOAZZDQRCGGP-GHCJXIJMSA-N 0.000 description 1
- PLOKOIJSGCISHE-BYULHYEWSA-N Asp-Val-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O PLOKOIJSGCISHE-BYULHYEWSA-N 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 238000011537 Coomassie blue staining Methods 0.000 description 1
- LPBUBIHAVKXUOT-FXQIFTODSA-N Cys-Val-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CS)N LPBUBIHAVKXUOT-FXQIFTODSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- RMOCFPBLHAOTDU-ACZMJKKPSA-N Gln-Asn-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O RMOCFPBLHAOTDU-ACZMJKKPSA-N 0.000 description 1
- GMGKDVVBSVVKCT-NUMRIWBASA-N Gln-Asn-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GMGKDVVBSVVKCT-NUMRIWBASA-N 0.000 description 1
- DRDSQGHKTLSNEA-GLLZPBPUSA-N Gln-Glu-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O DRDSQGHKTLSNEA-GLLZPBPUSA-N 0.000 description 1
- ICDIMQAMJGDHSE-GUBZILKMSA-N Gln-His-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(O)=O ICDIMQAMJGDHSE-GUBZILKMSA-N 0.000 description 1
- GQZDDFRXSDGUNG-YVNDNENWSA-N Gln-Ile-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(O)=O GQZDDFRXSDGUNG-YVNDNENWSA-N 0.000 description 1
- MLSKFHLRFVGNLL-WDCWCFNPSA-N Gln-Leu-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MLSKFHLRFVGNLL-WDCWCFNPSA-N 0.000 description 1
- CELXWPDNIGWCJN-WDCWCFNPSA-N Gln-Lys-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CELXWPDNIGWCJN-WDCWCFNPSA-N 0.000 description 1
- GQTNWYFWSUFFRA-KKUMJFAQSA-N Gln-Met-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O GQTNWYFWSUFFRA-KKUMJFAQSA-N 0.000 description 1
- OZEQPCDLCDRCGY-SOUVJXGZSA-N Gln-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CCC(=O)N)N)C(=O)O OZEQPCDLCDRCGY-SOUVJXGZSA-N 0.000 description 1
- HLRLXVPRJJITSK-IFFSRLJSSA-N Gln-Thr-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O HLRLXVPRJJITSK-IFFSRLJSSA-N 0.000 description 1
- BUZMZDDKFCSKOT-CIUDSAMLSA-N Glu-Glu-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O BUZMZDDKFCSKOT-CIUDSAMLSA-N 0.000 description 1
- ITBHUUMCJJQUSC-LAEOZQHASA-N Glu-Ile-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O ITBHUUMCJJQUSC-LAEOZQHASA-N 0.000 description 1
- ZGEJRLJEAMPEDV-SRVKXCTJSA-N Glu-Lys-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(=O)O)N ZGEJRLJEAMPEDV-SRVKXCTJSA-N 0.000 description 1
- KXTAGESXNQEZKB-DZKIICNBSA-N Glu-Phe-Val Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C(C)C)C(O)=O)CC1=CC=CC=C1 KXTAGESXNQEZKB-DZKIICNBSA-N 0.000 description 1
- NNQDRRUXFJYCCJ-NHCYSSNCSA-N Glu-Pro-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O NNQDRRUXFJYCCJ-NHCYSSNCSA-N 0.000 description 1
- MLILEEIVMRUYBX-NHCYSSNCSA-N Glu-Val-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O MLILEEIVMRUYBX-NHCYSSNCSA-N 0.000 description 1
- UGVQELHRNUDMAA-BYPYZUCNSA-N Gly-Ala-Gly Chemical compound [NH3+]CC(=O)N[C@@H](C)C(=O)NCC([O-])=O UGVQELHRNUDMAA-BYPYZUCNSA-N 0.000 description 1
- QXPRJQPCFXMCIY-NKWVEPMBSA-N Gly-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN QXPRJQPCFXMCIY-NKWVEPMBSA-N 0.000 description 1
- LJPIRKICOISLKN-WHFBIAKZSA-N Gly-Ala-Ser Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O LJPIRKICOISLKN-WHFBIAKZSA-N 0.000 description 1
- DWUKOTKSTDWGAE-BQBZGAKWSA-N Gly-Asn-Arg Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DWUKOTKSTDWGAE-BQBZGAKWSA-N 0.000 description 1
- JUGQPPOVWXSPKJ-RYUDHWBXSA-N Gly-Gln-Phe Chemical compound [H]NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JUGQPPOVWXSPKJ-RYUDHWBXSA-N 0.000 description 1
- MOJKRXIRAZPZLW-WDSKDSINSA-N Gly-Glu-Ala Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O MOJKRXIRAZPZLW-WDSKDSINSA-N 0.000 description 1
- BUEFQXUHTUZXHR-LURJTMIESA-N Gly-Gly-Pro zwitterion Chemical compound NCC(=O)NCC(=O)N1CCC[C@H]1C(O)=O BUEFQXUHTUZXHR-LURJTMIESA-N 0.000 description 1
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 1
- UHPAZODVFFYEEL-QWRGUYRKSA-N Gly-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)CN UHPAZODVFFYEEL-QWRGUYRKSA-N 0.000 description 1
- TVUWMSBGMVAHSJ-KBPBESRZSA-N Gly-Leu-Phe Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 TVUWMSBGMVAHSJ-KBPBESRZSA-N 0.000 description 1
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 1
- BXICSAQLIHFDDL-YUMQZZPRSA-N Gly-Lys-Asn Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O BXICSAQLIHFDDL-YUMQZZPRSA-N 0.000 description 1
- WMGHDYWNHNLGBV-ONGXEEELSA-N Gly-Phe-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 WMGHDYWNHNLGBV-ONGXEEELSA-N 0.000 description 1
- IGOYNRWLWHWAQO-JTQLQIEISA-N Gly-Phe-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 IGOYNRWLWHWAQO-JTQLQIEISA-N 0.000 description 1
- GGAPHLIUUTVYMX-QWRGUYRKSA-N Gly-Phe-Ser Chemical compound OC[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)C[NH3+])CC1=CC=CC=C1 GGAPHLIUUTVYMX-QWRGUYRKSA-N 0.000 description 1
- IRJWAYCXIYUHQE-WHFBIAKZSA-N Gly-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)CN IRJWAYCXIYUHQE-WHFBIAKZSA-N 0.000 description 1
- CSMYMGFCEJWALV-WDSKDSINSA-N Gly-Ser-Gln Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(N)=O CSMYMGFCEJWALV-WDSKDSINSA-N 0.000 description 1
- CQMFNTVQVLQRLT-JHEQGTHGSA-N Gly-Thr-Gln Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O CQMFNTVQVLQRLT-JHEQGTHGSA-N 0.000 description 1
- ONSARSFSJHTMFJ-STQMWFEESA-N Gly-Trp-Ser Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CO)C(O)=O ONSARSFSJHTMFJ-STQMWFEESA-N 0.000 description 1
- MREVELMMFOLESM-HOCLYGCPSA-N Gly-Trp-Val Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C(C)C)C(O)=O MREVELMMFOLESM-HOCLYGCPSA-N 0.000 description 1
- OCRQUYDOYKCOQG-IRXDYDNUSA-N Gly-Tyr-Phe Chemical compound C([C@H](NC(=O)CN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 OCRQUYDOYKCOQG-IRXDYDNUSA-N 0.000 description 1
- ZVXMEWXHFBYJPI-LSJOCFKGSA-N Gly-Val-Ile Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O ZVXMEWXHFBYJPI-LSJOCFKGSA-N 0.000 description 1
- KSOBNUBCYHGUKH-UWVGGRQHSA-N Gly-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN KSOBNUBCYHGUKH-UWVGGRQHSA-N 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- DRKZDEFADVYTLU-AVGNSLFASA-N His-Val-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(O)=O DRKZDEFADVYTLU-AVGNSLFASA-N 0.000 description 1
- TZCGZYWNIDZZMR-UHFFFAOYSA-N Ile-Arg-Ala Natural products CCC(C)C(N)C(=O)NC(C(=O)NC(C)C(O)=O)CCCN=C(N)N TZCGZYWNIDZZMR-UHFFFAOYSA-N 0.000 description 1
- RPZFUIQVAPZLRH-GHCJXIJMSA-N Ile-Asp-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C)C(=O)O)N RPZFUIQVAPZLRH-GHCJXIJMSA-N 0.000 description 1
- RGSOCXHDOPQREB-ZPFDUUQYSA-N Ile-Asp-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N RGSOCXHDOPQREB-ZPFDUUQYSA-N 0.000 description 1
- DMZOUKXXHJQPTL-GRLWGSQLSA-N Ile-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N DMZOUKXXHJQPTL-GRLWGSQLSA-N 0.000 description 1
- DVRDRICMWUSCBN-UKJIMTQDSA-N Ile-Gln-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N DVRDRICMWUSCBN-UKJIMTQDSA-N 0.000 description 1
- PHIXPNQDGGILMP-YVNDNENWSA-N Ile-Glu-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N PHIXPNQDGGILMP-YVNDNENWSA-N 0.000 description 1
- NYEYYMLUABXDMC-NHCYSSNCSA-N Ile-Gly-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)O)N NYEYYMLUABXDMC-NHCYSSNCSA-N 0.000 description 1
- RMNMUUCYTMLWNA-ZPFDUUQYSA-N Ile-Lys-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)O)C(=O)O)N RMNMUUCYTMLWNA-ZPFDUUQYSA-N 0.000 description 1
- FQYQMFCIJNWDQZ-CYDGBPFRSA-N Ile-Pro-Pro Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 FQYQMFCIJNWDQZ-CYDGBPFRSA-N 0.000 description 1
- KTNGVMMGIQWIDV-OSUNSFLBSA-N Ile-Pro-Thr Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O KTNGVMMGIQWIDV-OSUNSFLBSA-N 0.000 description 1
- XMYURPUVJSKTMC-KBIXCLLPSA-N Ile-Ser-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N XMYURPUVJSKTMC-KBIXCLLPSA-N 0.000 description 1
- JJQQGCMKLOEGAV-OSUNSFLBSA-N Ile-Thr-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)O)N JJQQGCMKLOEGAV-OSUNSFLBSA-N 0.000 description 1
- BZUOLKFQVVBTJY-SLBDDTMCSA-N Ile-Trp-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC(=O)N)C(=O)O)N BZUOLKFQVVBTJY-SLBDDTMCSA-N 0.000 description 1
- DTPGSUQHUMELQB-GVARAGBVSA-N Ile-Tyr-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 DTPGSUQHUMELQB-GVARAGBVSA-N 0.000 description 1
- APQYGMBHIVXFML-OSUNSFLBSA-N Ile-Val-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N APQYGMBHIVXFML-OSUNSFLBSA-N 0.000 description 1
- IBMVEYRWAWIOTN-UHFFFAOYSA-N L-Leucyl-L-Arginyl-L-Proline Natural products CC(C)CC(N)C(=O)NC(CCCN=C(N)N)C(=O)N1CCCC1C(O)=O IBMVEYRWAWIOTN-UHFFFAOYSA-N 0.000 description 1
- DBVWMYGBVFCRBE-CIUDSAMLSA-N Leu-Asn-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O DBVWMYGBVFCRBE-CIUDSAMLSA-N 0.000 description 1
- OIARJGNVARWKFP-YUMQZZPRSA-N Leu-Asn-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O OIARJGNVARWKFP-YUMQZZPRSA-N 0.000 description 1
- VDIARPPNADFEAV-WEDXCCLWSA-N Leu-Thr-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O VDIARPPNADFEAV-WEDXCCLWSA-N 0.000 description 1
- FDBTVENULFNTAL-XQQFMLRXSA-N Leu-Val-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N FDBTVENULFNTAL-XQQFMLRXSA-N 0.000 description 1
- RVOMPSJXSRPFJT-DCAQKATOSA-N Lys-Ala-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RVOMPSJXSRPFJT-DCAQKATOSA-N 0.000 description 1
- KWUKZRFFKPLUPE-HJGDQZAQSA-N Lys-Asp-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KWUKZRFFKPLUPE-HJGDQZAQSA-N 0.000 description 1
- MRWXLRGAFDOILG-DCAQKATOSA-N Lys-Gln-Gln Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MRWXLRGAFDOILG-DCAQKATOSA-N 0.000 description 1
- WWEWGPOLIJXGNX-XUXIUFHCSA-N Lys-Met-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCCN)N WWEWGPOLIJXGNX-XUXIUFHCSA-N 0.000 description 1
- VHTOGMKQXXJOHG-RHYQMDGZSA-N Lys-Thr-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O VHTOGMKQXXJOHG-RHYQMDGZSA-N 0.000 description 1
- DNDVVILEHVMWIS-LPEHRKFASA-N Met-Asp-Pro Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N DNDVVILEHVMWIS-LPEHRKFASA-N 0.000 description 1
- GPVLSVCBKUCEBI-KKUMJFAQSA-N Met-Gln-Phe Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O GPVLSVCBKUCEBI-KKUMJFAQSA-N 0.000 description 1
- LRALLISKBZNSKN-BQBZGAKWSA-N Met-Gly-Ser Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O LRALLISKBZNSKN-BQBZGAKWSA-N 0.000 description 1
- AEQVPPGEJJBFEE-CYDGBPFRSA-N Met-Ile-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AEQVPPGEJJBFEE-CYDGBPFRSA-N 0.000 description 1
- MIAZEQZXAFTCCG-UBHSHLNASA-N Met-Phe-Ala Chemical compound CSCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=CC=C1 MIAZEQZXAFTCCG-UBHSHLNASA-N 0.000 description 1
- TWEWRDAAIYBJTO-ULQDDVLXSA-N Met-Tyr-His Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N TWEWRDAAIYBJTO-ULQDDVLXSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- LSXGADJXBDFXQU-DLOVCJGASA-N Phe-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=CC=C1 LSXGADJXBDFXQU-DLOVCJGASA-N 0.000 description 1
- CDNPIRSCAFMMBE-SRVKXCTJSA-N Phe-Asn-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O CDNPIRSCAFMMBE-SRVKXCTJSA-N 0.000 description 1
- RTUWVJVJSMOGPL-KKUMJFAQSA-N Phe-Met-Glu Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N RTUWVJVJSMOGPL-KKUMJFAQSA-N 0.000 description 1
- ABEFOXGAIIJDCL-SFJXLCSZSA-N Phe-Thr-Trp Chemical compound C([C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=CC=C1 ABEFOXGAIIJDCL-SFJXLCSZSA-N 0.000 description 1
- KIQUCMUULDXTAZ-HJOGWXRNSA-N Phe-Tyr-Tyr Chemical compound N[C@@H](Cc1ccccc1)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N[C@@H](Cc1ccc(O)cc1)C(O)=O KIQUCMUULDXTAZ-HJOGWXRNSA-N 0.000 description 1
- XBCOOBCTVMMQSC-BVSLBCMMSA-N Phe-Val-Trp Chemical compound C([C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=CC=C1 XBCOOBCTVMMQSC-BVSLBCMMSA-N 0.000 description 1
- 206010035148 Plague Diseases 0.000 description 1
- XWYXZPHPYKRYPA-GMOBBJLQSA-N Pro-Asn-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O XWYXZPHPYKRYPA-GMOBBJLQSA-N 0.000 description 1
- FUVBEZJCRMHWEM-FXQIFTODSA-N Pro-Asn-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O FUVBEZJCRMHWEM-FXQIFTODSA-N 0.000 description 1
- JARJPEMLQAWNBR-GUBZILKMSA-N Pro-Asp-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O JARJPEMLQAWNBR-GUBZILKMSA-N 0.000 description 1
- KPDRZQUWJKTMBP-DCAQKATOSA-N Pro-Asp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@@H]1CCCN1 KPDRZQUWJKTMBP-DCAQKATOSA-N 0.000 description 1
- QVIZLAUEAMQKGS-GUBZILKMSA-N Pro-Asp-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@@H]1CCCN1 QVIZLAUEAMQKGS-GUBZILKMSA-N 0.000 description 1
- ODPIUQVTULPQEP-CIUDSAMLSA-N Pro-Gln-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@@H]1CCCN1 ODPIUQVTULPQEP-CIUDSAMLSA-N 0.000 description 1
- FISHYTLIMUYTQY-GUBZILKMSA-N Pro-Gln-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1 FISHYTLIMUYTQY-GUBZILKMSA-N 0.000 description 1
- WGAQWMRJUFQXMF-ZPFDUUQYSA-N Pro-Gln-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WGAQWMRJUFQXMF-ZPFDUUQYSA-N 0.000 description 1
- LANQLYHLMYDWJP-SRVKXCTJSA-N Pro-Gln-Lys Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O LANQLYHLMYDWJP-SRVKXCTJSA-N 0.000 description 1
- FEPSEIDIPBMIOS-QXEWZRGKSA-N Pro-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 FEPSEIDIPBMIOS-QXEWZRGKSA-N 0.000 description 1
- FKLSMYYLJHYPHH-UWVGGRQHSA-N Pro-Gly-Leu Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O FKLSMYYLJHYPHH-UWVGGRQHSA-N 0.000 description 1
- MCWHYUWXVNRXFV-RWMBFGLXSA-N Pro-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2 MCWHYUWXVNRXFV-RWMBFGLXSA-N 0.000 description 1
- KIDXAAQVMNLJFQ-KZVJFYERSA-N Pro-Thr-Ala Chemical compound C[C@@H](O)[C@H](NC(=O)[C@@H]1CCCN1)C(=O)N[C@@H](C)C(O)=O KIDXAAQVMNLJFQ-KZVJFYERSA-N 0.000 description 1
- DCHQYSOGURGJST-FJXKBIBVSA-N Pro-Thr-Gly Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O DCHQYSOGURGJST-FJXKBIBVSA-N 0.000 description 1
- GZNYIXWOIUFLGO-ZJDVBMNYSA-N Pro-Thr-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZNYIXWOIUFLGO-ZJDVBMNYSA-N 0.000 description 1
- WTWGOQRNRFHFQD-JBDRJPRFSA-N Ser-Ala-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WTWGOQRNRFHFQD-JBDRJPRFSA-N 0.000 description 1
- OOKCGAYXSNJBGQ-ZLUOBGJFSA-N Ser-Asn-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O OOKCGAYXSNJBGQ-ZLUOBGJFSA-N 0.000 description 1
- SWIQQMYVHIXPEK-FXQIFTODSA-N Ser-Cys-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O SWIQQMYVHIXPEK-FXQIFTODSA-N 0.000 description 1
- UQFYNFTYDHUIMI-WHFBIAKZSA-N Ser-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CO UQFYNFTYDHUIMI-WHFBIAKZSA-N 0.000 description 1
- BPMRXBZYPGYPJN-WHFBIAKZSA-N Ser-Gly-Asn Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O BPMRXBZYPGYPJN-WHFBIAKZSA-N 0.000 description 1
- RIAKPZVSNBBNRE-BJDJZHNGSA-N Ser-Ile-Leu Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O RIAKPZVSNBBNRE-BJDJZHNGSA-N 0.000 description 1
- FPCGZYMRFFIYIH-CIUDSAMLSA-N Ser-Lys-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O FPCGZYMRFFIYIH-CIUDSAMLSA-N 0.000 description 1
- QMCDMHWAKMUGJE-IHRRRGAJSA-N Ser-Phe-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O QMCDMHWAKMUGJE-IHRRRGAJSA-N 0.000 description 1
- ADJDNJCSPNFFPI-FXQIFTODSA-N Ser-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO ADJDNJCSPNFFPI-FXQIFTODSA-N 0.000 description 1
- NUEHQDHDLDXCRU-GUBZILKMSA-N Ser-Pro-Arg Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O NUEHQDHDLDXCRU-GUBZILKMSA-N 0.000 description 1
- OLKICIBQRVSQMA-SRVKXCTJSA-N Ser-Ser-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OLKICIBQRVSQMA-SRVKXCTJSA-N 0.000 description 1
- SQHKXWODKJDZRC-LKXGYXEUSA-N Ser-Thr-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O SQHKXWODKJDZRC-LKXGYXEUSA-N 0.000 description 1
- ZSDXEKUKQAKZFE-XAVMHZPKSA-N Ser-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N)O ZSDXEKUKQAKZFE-XAVMHZPKSA-N 0.000 description 1
- ZKOKTQPHFMRSJP-YJRXYDGGSA-N Ser-Thr-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZKOKTQPHFMRSJP-YJRXYDGGSA-N 0.000 description 1
- VEVYMLNYMULSMS-AVGNSLFASA-N Ser-Tyr-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O VEVYMLNYMULSMS-AVGNSLFASA-N 0.000 description 1
- HAYADTTXNZFUDM-IHRRRGAJSA-N Ser-Tyr-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O HAYADTTXNZFUDM-IHRRRGAJSA-N 0.000 description 1
- UKKROEYWYIHWBD-ZKWXMUAHSA-N Ser-Val-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O UKKROEYWYIHWBD-ZKWXMUAHSA-N 0.000 description 1
- HSWXBJCBYSWBPT-GUBZILKMSA-N Ser-Val-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)C(O)=O HSWXBJCBYSWBPT-GUBZILKMSA-N 0.000 description 1
- SWIKDOUVROTZCW-GCJQMDKQSA-N Thr-Asn-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C)C(=O)O)N)O SWIKDOUVROTZCW-GCJQMDKQSA-N 0.000 description 1
- YLXAMFZYJTZXFH-OLHMAJIHSA-N Thr-Asn-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O YLXAMFZYJTZXFH-OLHMAJIHSA-N 0.000 description 1
- JHBHMCMKSPXRHV-NUMRIWBASA-N Thr-Asn-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O JHBHMCMKSPXRHV-NUMRIWBASA-N 0.000 description 1
- PQLXHSACXPGWPD-GSSVUCPTSA-N Thr-Asn-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PQLXHSACXPGWPD-GSSVUCPTSA-N 0.000 description 1
- JEDIEMIJYSRUBB-FOHZUACHSA-N Thr-Asp-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O JEDIEMIJYSRUBB-FOHZUACHSA-N 0.000 description 1
- ODSAPYVQSLDRSR-LKXGYXEUSA-N Thr-Cys-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(O)=O ODSAPYVQSLDRSR-LKXGYXEUSA-N 0.000 description 1
- GCXFWAZRHBRYEM-NUMRIWBASA-N Thr-Gln-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O GCXFWAZRHBRYEM-NUMRIWBASA-N 0.000 description 1
- SHOMROOOQBDGRL-JHEQGTHGSA-N Thr-Glu-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O SHOMROOOQBDGRL-JHEQGTHGSA-N 0.000 description 1
- LHEZGZQRLDBSRR-WDCWCFNPSA-N Thr-Glu-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LHEZGZQRLDBSRR-WDCWCFNPSA-N 0.000 description 1
- KZSYAEWQMJEGRZ-RHYQMDGZSA-N Thr-Leu-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O KZSYAEWQMJEGRZ-RHYQMDGZSA-N 0.000 description 1
- WTMPKZWHRCMMMT-KZVJFYERSA-N Thr-Pro-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O WTMPKZWHRCMMMT-KZVJFYERSA-N 0.000 description 1
- MXDOAJQRJBMGMO-FJXKBIBVSA-N Thr-Pro-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O MXDOAJQRJBMGMO-FJXKBIBVSA-N 0.000 description 1
- VTMGKRABARCZAX-OSUNSFLBSA-N Thr-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)[C@@H](C)O VTMGKRABARCZAX-OSUNSFLBSA-N 0.000 description 1
- AAZOYLQUEQRUMZ-GSSVUCPTSA-N Thr-Thr-Asn Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(N)=O AAZOYLQUEQRUMZ-GSSVUCPTSA-N 0.000 description 1
- BBPCSGKKPJUYRB-UVOCVTCTSA-N Thr-Thr-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O BBPCSGKKPJUYRB-UVOCVTCTSA-N 0.000 description 1
- ZESGVALRVJIVLZ-VFCFLDTKSA-N Thr-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@@H]1C(=O)O)N)O ZESGVALRVJIVLZ-VFCFLDTKSA-N 0.000 description 1
- QGVBFDIREUUSHX-IFFSRLJSSA-N Thr-Val-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O QGVBFDIREUUSHX-IFFSRLJSSA-N 0.000 description 1
- KPMIQCXJDVKWKO-IFFSRLJSSA-N Thr-Val-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KPMIQCXJDVKWKO-IFFSRLJSSA-N 0.000 description 1
- PNHABSVRPFBUJY-UMPQAUOISA-N Trp-Arg-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N)O PNHABSVRPFBUJY-UMPQAUOISA-N 0.000 description 1
- MHNHRNHJMXAVHZ-AAEUAGOBSA-N Trp-Asn-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)O)N MHNHRNHJMXAVHZ-AAEUAGOBSA-N 0.000 description 1
- RQKMZXSRILVOQZ-GMVOTWDCSA-N Trp-Tyr-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N RQKMZXSRILVOQZ-GMVOTWDCSA-N 0.000 description 1
- NZFCWALTLNFHHC-JYJNAYRXSA-N Tyr-Glu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NZFCWALTLNFHHC-JYJNAYRXSA-N 0.000 description 1
- QAYSODICXVZUIA-WLTAIBSBSA-N Tyr-Gly-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O QAYSODICXVZUIA-WLTAIBSBSA-N 0.000 description 1
- AOIZTZRWMSPPAY-KAOXEZKKSA-N Tyr-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)O AOIZTZRWMSPPAY-KAOXEZKKSA-N 0.000 description 1
- JHDZONWZTCKTJR-KJEVXHAQSA-N Tyr-Thr-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 JHDZONWZTCKTJR-KJEVXHAQSA-N 0.000 description 1
- ZLFHAAGHGQBQQN-AEJSXWLSSA-N Val-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N ZLFHAAGHGQBQQN-AEJSXWLSSA-N 0.000 description 1
- ZLFHAAGHGQBQQN-GUBZILKMSA-N Val-Ala-Pro Natural products CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O ZLFHAAGHGQBQQN-GUBZILKMSA-N 0.000 description 1
- CVUDMNSZAIZFAE-TUAOUCFPSA-N Val-Arg-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@@H]1C(=O)O)N CVUDMNSZAIZFAE-TUAOUCFPSA-N 0.000 description 1
- CVUDMNSZAIZFAE-UHFFFAOYSA-N Val-Arg-Pro Natural products NC(N)=NCCCC(NC(=O)C(N)C(C)C)C(=O)N1CCCC1C(O)=O CVUDMNSZAIZFAE-UHFFFAOYSA-N 0.000 description 1
- VUTHNLMCXKLLFI-LAEOZQHASA-N Val-Asp-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N VUTHNLMCXKLLFI-LAEOZQHASA-N 0.000 description 1
- NXRAUQGGHPCJIB-RCOVLWMOSA-N Val-Gly-Asn Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O NXRAUQGGHPCJIB-RCOVLWMOSA-N 0.000 description 1
- HGJRMXOWUWVUOA-GVXVVHGQSA-N Val-Leu-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N HGJRMXOWUWVUOA-GVXVVHGQSA-N 0.000 description 1
- BTWMICVCQLKKNR-DCAQKATOSA-N Val-Leu-Ser Chemical compound CC(C)[C@H]([NH3+])C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C([O-])=O BTWMICVCQLKKNR-DCAQKATOSA-N 0.000 description 1
- MBGFDZDWMDLXHQ-GUBZILKMSA-N Val-Met-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](C(C)C)N MBGFDZDWMDLXHQ-GUBZILKMSA-N 0.000 description 1
- SJRUJQFQVLMZFW-WPRPVWTQSA-N Val-Pro-Gly Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O SJRUJQFQVLMZFW-WPRPVWTQSA-N 0.000 description 1
- YQYFYUSYEDNLSD-YEPSODPASA-N Val-Thr-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O YQYFYUSYEDNLSD-YEPSODPASA-N 0.000 description 1
- WUFHZIRMAZZWRS-OSUNSFLBSA-N Val-Thr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](C(C)C)N WUFHZIRMAZZWRS-OSUNSFLBSA-N 0.000 description 1
- JVGDAEKKZKKZFO-RCWTZXSCSA-N Val-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)N)O JVGDAEKKZKKZFO-RCWTZXSCSA-N 0.000 description 1
- 108010087302 Viral Structural Proteins Proteins 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 108010087049 alanyl-alanyl-prolyl-valine Proteins 0.000 description 1
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 108010041407 alanylaspartic acid Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 238000003277 amino acid sequence analysis Methods 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 108010068380 arginylarginine Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940031416 bivalent vaccine Drugs 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000000234 capsid Anatomy 0.000 description 1
- 230000000120 cytopathologic effect Effects 0.000 description 1
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000011013 endotoxin removal Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010001064 glycyl-glycyl-glycyl-glycine Proteins 0.000 description 1
- 108010051307 glycyl-glycyl-proline Proteins 0.000 description 1
- 108010077435 glycyl-phenylalanyl-glycine Proteins 0.000 description 1
- 108010017446 glycyl-prolyl-arginyl-proline Proteins 0.000 description 1
- 108010010147 glycylglutamine Proteins 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 108010085325 histidylproline Proteins 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 230000016784 immunoglobulin production Effects 0.000 description 1
- 230000002480 immunoprotective effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 108010078274 isoleucylvaline Proteins 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010051673 leucyl-glycyl-phenylalanine Proteins 0.000 description 1
- 108010030617 leucyl-phenylalanyl-valine Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 108010016686 methionyl-alanyl-serine Proteins 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- MXHCPCSDRGLRER-UHFFFAOYSA-N pentaglycine Chemical compound NCC(=O)NCC(=O)NCC(=O)NCC(=O)NCC(O)=O MXHCPCSDRGLRER-UHFFFAOYSA-N 0.000 description 1
- 108010070409 phenylalanyl-glycyl-glycine Proteins 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 1
- 108010083476 phenylalanyltryptophan Proteins 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 1
- 108010029020 prolylglycine Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 108010005834 tyrosyl-alanyl-glycine Proteins 0.000 description 1
- IBIDRSSEHFLGSD-UHFFFAOYSA-N valinyl-arginine Natural products CC(C)C(N)C(=O)NC(C(O)=O)CCCN=C(N)N IBIDRSSEHFLGSD-UHFFFAOYSA-N 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/02—Bacterial antigens
- A61K39/102—Pasteurellales, e.g. Actinobacillus, Pasteurella; Haemophilus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/285—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Pasteurellaceae (F), e.g. Haemophilus influenza
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/70—Multivalent vaccine
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/14011—Baculoviridae
- C12N2710/14111—Nucleopolyhedrovirus, e.g. autographa californica nucleopolyhedrovirus
- C12N2710/14141—Use of virus, viral particle or viral elements as a vector
- C12N2710/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/16011—Caliciviridae
- C12N2770/16022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/16011—Caliciviridae
- C12N2770/16034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2800/00—Nucleic acids vectors
- C12N2800/10—Plasmid DNA
- C12N2800/103—Plasmid DNA for invertebrates
- C12N2800/105—Plasmid DNA for invertebrates for insects
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Virology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Molecular Biology (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Communicable Diseases (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Oncology (AREA)
- Immunology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明属生物医学技术领域，提供了一种双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原，所述重组抗原通过在分析巴氏杆菌保护性抗原PlpE表位组成的基础上，将PlpE表位集中分布的一段区域（长度合计70个氨基酸）分两段嵌合展示到兔出血症病毒VP60蛋白N端和C端而获得。免疫保护力实验表明该嵌合重组抗原对兔出血症病毒和巴氏杆菌均具有良好的免疫保护力。本发明仅耗费一份人工就可以制备出针对两种病原的抗原，简化了工艺，缩短了制备时间，显著降低了生产成本，为后期生产更低成本的兔病毒性出血症、兔巴氏杆菌病疫苗提供了基础。
Description
技术领域
本发明属生物医学技术领域，具体涉及一种嵌合了巴氏杆菌抗原表位的兔出血症病毒VP60重组抗原及其制备方法和应用。
背景技术
兔病毒性出血症，俗称兔瘟，是由兔出血症病毒( Rabbit hemorrhagic diseasevirus，RHDV) 引起的一种具有高度传染性、高发病率、高致死性的疾病，主要发生于 2 月龄以上成年兔。RHDV 衣壳蛋白 VP60 由 579 个氨基酸组成，是病毒衣壳组成的最基本单位。衣壳蛋白 VP60 是病毒的免疫保护性抗原，与诱导机体抗感染免疫直接相关，单独表达VP60蛋白能够自聚成病毒样颗粒，由其制成的疫苗已经用于兔病毒性出血症的预防。
兔巴氏杆菌病（Pasteurellosis）是主要由多杀性巴氏杆菌（Pasteurella multocida）引起的一种危害养兔业的主要细菌性疫病。各种年龄、品种的家兔对于多杀性巴氏杆菌均易感，尤其2-6月龄的家兔易感染发病，该病是造成9周龄到6月龄家兔死亡的主要疫病之一。目前已经有PlpE等保护性抗原报道，动物实验显示出良好的免疫保护效力。
目前已经有兔病毒性出血症、兔巴氏杆菌病二联灭活疫苗用于同时防控兔病毒性出血症和兔巴氏杆菌病。但是该疫苗存在着生产成本偏高等、对巴氏杆菌病的免疫保护效果并不理想等问题。传统兔病毒性出血症、兔巴氏杆菌病二联灭活疫苗的制备工艺是先分别制备兔病毒性出血症、兔巴氏杆菌灭活疫苗，再进行物理混合制成联苗。这种二联苗的生产过程需要经历两个疫苗的生产过程，耗费两份人工及物料、水电等，增加了生产成本。虽然近年来有使用基因工程亚单位疫苗（以VP60和PlpE等为抗原）取代传统细菌或病毒灭活疫苗的趋势，但是目前基因工程二联亚单位疫苗的生产工艺依然需要经历分别制备单苗然后物理混合的过程，生产成本并未显著降低。因而将不同抗原及其表位嵌合连接在一起构成新抗原的方案正成为新的联苗发展方向，这将使二联疫苗生产过程简化，有效降低生产成本。
本发明的目的是将巴氏杆菌保护性抗原PlpE的表位嵌合展示到兔出血症病毒VP60上，从而获得一种新的兔病毒性出血症、兔巴氏杆菌病二联亚单位疫苗所需抗原。
发明内容
本发明所解决的技术问题是：通过构建嵌合有巴氏杆菌保护性抗原PlpE抗原表位的兔出血症病毒VP60重组蛋白抗原，获得一种只耗费一份人工就能够生产出对兔出血症病毒和巴氏杆菌都具有免疫保护作用的重组蛋白抗原，从而应用于兔病毒性出血症、兔巴氏杆菌病二联疫苗制备
为了解决上述技术问题，本发明提供一种双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原，所述重组抗原包括兔出血症病毒VP60蛋白，且其N端和C端分别连接了PlpE up多肽片段和PlpE down多肽片段；其中，PlpE up多肽片段和PlpE down多肽片段源自巴氏杆菌PlpE蛋白抗原表位。
所述VP60蛋白N端和C端所嵌入的PlpE抗原表位氨基酸序列分别为SEQ ID NO:1和SEQ ID NO:2。
编码所述VP60蛋白N端和C端所嵌入的PlpE抗原表位核苷酸序列分别为SEQ IDNO:3和SEQ ID NO:4。
所述双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原氨基酸序列为SEQ ID NO:5；编码所述双位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原的核苷酸序列为SEQ ID NO:6。
此外，提供了一种重组抗原的表达载体，所述载体用于表达双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原，含有权利要求7所述核苷酸序列。
本发明所述双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原制备方法，其步骤如下：
(1)通过PCR扩增巴氏杆菌PlpE的第26位到第50位氨基酸所对应的DNA序列、第51位到第95位氨基酸对应的DNA序列;
(2)将步骤(1)得到PCR产物依次分别连接到VP60基因的5’端和3’端上，并将重组基因连接到pFastBac1克隆位点上,得到重组穿梭质粒载体;
(3)将步骤(2)得到的重组穿梭载体转化到DH10Bac宿主菌中，通过该菌的转座作用获得杆状病毒质粒；
（4）将重组杆状病毒质粒转入sf9细胞后获得嵌合了巴氏杆菌PlpE表位的兔出血症病毒VP60重组蛋白抗原。
上述双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原在制备预防兔病毒性出血症和兔巴氏杆菌病疫苗方面的应用。
本发明的有益效果是：与现有技术相比，尽管目前现有技术采用兔病毒性出血症、兔巴氏杆菌二联灭活疫苗或基因工程亚单位疫苗来同时防控兔病毒性出血症、兔巴氏杆菌病，但是传统二联苗的制备工艺是先分别制备两种疫苗再进行物理混合，二联苗的生产过程需要经历两个疫苗的生产过程，耗费两份人工及物料、水电等，增加了生产成本，且带来不同组分相互干扰的风险。虽然理论上兔出血症病毒结构蛋白VP60的N端和C端具有展示外源表位的能力，但既要尽可能多的展示外源表位（特别是亲缘关系比较远的细菌抗原表位），又要保证自身表位不受影响则面临较大的困难（目前技术条件下VP60蛋白N端和C端最长仅能展示合计为40个氨基酸左右的外源动物病毒表位）。本发明在分析巴氏杆菌保护性抗原PlpE表位组成的基础上，通过将PlpE表位集中分布的一段区域（长度合计70个氨基酸）分两段嵌合展示到兔出血症病毒VP60蛋白N端和C端，意外发现该重组抗原仍然具有血凝活性（即能够形成病毒样颗粒，兔出血症病毒病毒中和表位未受影响）。免疫保护力实验结果表明该重组抗原既对巴氏杆菌有免疫保护作用，又对兔出血症病毒具有免疫保护作用。
总之，本发明通过构建嵌合了巴氏杆菌保护性抗原PlpE表位的兔出血症病毒Vp60重组蛋白，获得了对兔出血症病毒和巴氏杆菌都具有免疫保护作用的重组抗原，为后期生产更低成本的兔病毒性出血症、兔巴氏杆菌病疫苗提供了基础。
附图说明
图1为PlpE的氨基酸残基构成B细胞表位能力预测结果(使用在线软件Bepipred-2.0（http://tools.immuneepitope.org/bcell/），预测分值高于0.5构成B细胞表位的几率较大)；
图2 为PlpE表位双位点嵌合VP60重组抗原的重组穿梭质粒载体构建过程示意图；
图3为PlpE表位嵌合VP60重组蛋白VP60-PlpE的表达情况，其中M表示分子大小标识； VP60-PlpE为杆状病毒-昆虫细胞表达系统表达的VP60-PlpE重组蛋白（68 kDa）；VP60表示杆状病毒-昆虫细胞表达系统表达的兔出血症病毒VP60蛋白（60 kDa）；
图4为本发明所述PlpE表位嵌合VP60重组蛋白后VP60-PlpE的血凝特性，左图为VP60-PlpE对人O型红细胞的凝集；右图为对照，即野生型杆状病毒培养物对人O型红细胞的凝集；
图5为本发明所述PlpE表位嵌合VP60重组蛋白VP60-PlpE与PlpE高免血清的反应情况，其中M表示分子大小标识；VP60-PlpE为杆状病毒-昆虫细胞表达系统表达的VP60-PlpE重组蛋白；VP60表示杆状病毒-昆虫细胞表达系统表达的兔出血症病毒VP60蛋白；
图6为免疫保护力实验中特异性抗体产生情况（A）和巴氏杆菌攻毒后小鼠的存活情况（B）
图7免疫保护力实验中兔出血症病毒攻毒（A）和巴氏杆菌攻毒（B）后家兔的存活情况。
具体实施方式
为了使本领域的技术人员更好地理解本发明的技术方案，下面结合具体实施例对本发明作进一步的详细说明。
实施例1：制备PlpE表位嵌合的VP60重组蛋白
(1)质粒、菌种及培养载体质粒pFastBac1、pFastBac1-VP60 由本实验室构建并保存; E．coli DH5α 感受态购买自南京诺唯赞生物技术有限公司; E．coli DH10 Bac 感受态购自上海唯地生物科技有限公司; 多杀性巴氏杆菌C51-17株由本实验室保存提供；sf9昆虫细胞由本实验室保存培养。
蛋白质氨基酸序列分析以Genbank公布的多杀性巴氏杆菌C51-17株的全基因组序列中的PlpE基因序列为参考序列进行分析。使用Bepipred-2.0（http://tools.immuneepitope.org/bcell/）确定PlpE的B细胞表位分布情况。
上述PlpE的B细胞表位预测结果如图1所示:PlpE一级结构共包含336个氨基酸残基。选择Bepipred-2.0软件默认的0.5作为氨基酸残基构成表位的阈值，预测结果表明PlpE的表位主要分布于N端，特别是第26位-105位氨基酸残基序列。因此本发明选择了将N端26位到95位氨基酸分段连接到兔出血症病毒VP60的N端和C端获得表位嵌合VP60，即本发明所述用作兔病毒性出血症、兔巴氏杆菌病疫苗制备的重组蛋白抗原,其氨基酸序列为SEQ IDNO :5.
(3)重组杆粒构建以多杀性巴氏杆菌C51-17株的PlpE基因序列为参考序列，设计引物PlpEupF：5’-aggcatgcggtaccaagcttatgTGTAGCGGTGGTGGCGGT-3’和PlpEupR：5’-gggctttgccctcATTTGATTGAACCGGTTGTGCT-3’用于扩增PlpE的第26位到第50位氨基酸所对应的DNA片段，PlpEdownF：5’-ggcttttcttatgtcACTCCAATCAAACATCCTATGACTAATAG-3’和PlpEdownR：5’-atcctctagtacttctcgacctaTTTTTCTTGTTCTAGAGGGGCTTG-3’扩增PlpE第51位到95位氨基酸所对应的DNA片段。提取巴氏杆菌C51-17菌株基因组并进行PCR扩增目的PlpE基因片段。使用引物VP60F 5’-atcaaatGAGGGCAAAGCCCGCACA-3’和VP60R 5’ggagtGACATAAGAAAAGCCATTGGTTGTG3’扩增pFastBac1-VP60质粒载体制备VP60基因片段。使用引物pFastBacF5’-GTCGAGAAGTACTAGAGGATCATAATCAGCC-3’和pFastBacR5’-AAGCTTGGTACCGCATGCC-3’PCR扩增pFastBac1质粒载体制备线性化pFastBac1线性化载体。按照同源重组克隆试剂盒（诺唯赞，南京）说明书中的操作将PlpE第26-50位、第51-95位氨基酸对应的基因片段分别连接到VP60基因的5’端和3’端上，然后将阳性重组质粒转入DH5α宿主菌。挑取阳性克隆PCR鉴定正确后送北京擎科新业有限公司南京分部测序，鉴定所克隆基因是否完整及是否正确连接到载体上,测序结果显示目的基因片段已经正确连接到载体上。提取阳性克隆质粒，按照E． coli DH10Bac 感受态细胞说明书中的操作，将重组质粒转化到E． coli DH10Bac 感受态细胞中并通过蓝白斑筛选和 PCR 鉴定获得确定阳性克隆，提取重组杆粒Bacmid-VP60-PlpE。所构建的重组杆粒如图2所示。
转染、表达与鉴定将Bacmid-VP60-PlpE通过脂质体 LipofectaminTM3000 转染至对数生长期的Sf9 昆虫细胞。转染后每 12 h 观察 1 次，细胞病变明显时收集细胞及上清液，作为重组杆状病毒原液，4 ℃保存。将第 1 代病毒液反复冻融 3 次后以 1% 体积比接种 Sf9 细胞，进行传代，得到第 2 代重组病毒。重复上述操作获得3代毒后将重组杆状病毒接种到对数期的悬浮sf9细胞中进行转瓶培养，培养约100h后收集培养物。反复冻融培养物3次后获得VP60-PlpE重组蛋白，进行SDS-PAGE，考马斯亮蓝染色后进行观察，样品中存在约68kDa的条带（图3），符合VP60-PlpE理论预测大小。
实施例2 双位点嵌合VP60-PlpE重组蛋白的血凝试验
在50孔U型板上两个孔内分别加入VP60-PlpE和野生型杆状病毒培养物，每孔 50μL。然后每孔加入 50 μL 1%人 O 型红细胞悬液，4 ℃作用30 min 后，观察血凝情况。结果表明VP60-PlpE重组蛋白存在明显的血凝现象（图4），即认为VP60-PlpE重组蛋白仍然可以形成兔出血症病毒样颗粒，有效展现兔出血症病毒中和表位。
实施例3双位点嵌合VP60-PlpE重组蛋白与PlpE高免血清反应
将VP60-PlpE重组蛋白和VP60重组蛋白（对照）进行SDS-PAGE凝胶电泳后将蛋白转印到NC膜上。经过5%的脱脂乳封闭后，加入本实验室保存的1/500稀释的小鼠抗PlpE血清，37℃孵育1 h；PBST漂洗5遍，加入1/10000稀释的HRP偶联的羊抗鼠IgG，37℃孵育1 h； PBST漂洗5遍，进行ECL显色。Western blot结果显示，VP60-PlpE泳道存在一条68 kDa左右的条带（图5），但作为对照的VP60泳道则不存在符合预期大小的条带，这就表明VP60-PlpE重组蛋白能够与小鼠抗PlpE血清发生特异性反应，即PlpE部分免疫原性片段顺利整合到VP60上。
实施例4免疫保护力实验
(1)小鼠免疫保护力实验取20 只 4-6 周龄ICR雌鼠，随机分成2组，1组对照，另一组实验，每组10只。实验组免疫VP60-PlpE重组蛋白：200μg/μl重组蛋白中加入20%体积铝胶后，腹腔注射免疫小鼠，每只100μl。21天后加强免疫，剂量、方式等与初免相同。阴性对照组：PBS代替重组蛋白，使用的佐剂与实验组的佐剂相同。加强免疫10天后对所有小鼠尾静脉采血，确认已经产生高水平特异性抗体（图6A）以后，使用巴氏杆菌C51-17株对小鼠皮下注射攻毒，每只50CFU左右（约5×LD50）。攻毒后小鼠的存活情况如图6B所示，对照组仅有2只小鼠存活，实验组小鼠存活8只。这就表明VP60-PlpE使小鼠具有了对巴氏杆菌攻毒的免疫保护力。
(2)家兔免疫保护力实验选取60只2-3月龄健康易感家兔，随机分成6组，2组对照，另4组实验，每组10只。两个实验组免疫VP60-PlpE重组蛋白组：200mg/ml重组蛋白中加入20%体积的铝胶佐剂后皮下注射免疫家兔，每只1ml；另两个实验组免疫VP60和PlpE重组蛋白的物理混合物（Mixture），其中包含VP60重组蛋白和PlpE重组蛋白各100mg/ml，加入20%体积的铝胶佐剂后免疫家兔。剩余两组作为阴性对照组：免疫PBS代替重组蛋白，使用的佐剂与实验组的佐剂相同。免疫3周后使用兔出血症病毒皖阜株对VP60-PlpE嵌合抗原免疫组、物理混合Mixture组和一组对照组攻毒，100×LD50/只，连续7天观察家兔存活情况。免疫3周后使用巴氏杆菌C51-17株对剩余VP60-PlpE嵌合抗原免疫组、物理混合mixture组和另一组对照组攻毒，100CFU（10×MLD）/只，连续7天观察家兔存活情况。兔出血症病毒攻毒后实验组家（免疫VP60-PlpE嵌合抗原重组蛋白组和免疫两种蛋白物理混合物组）兔存活率100%，对照组全部死亡（存活率0/10，图7A）。巴氏杆菌攻毒后实验组家兔存活率100%，而对照组仅存活20%（图7B）。这些实验结果表明VP60-PlpE重组蛋白的免疫能使家兔同时对兔出血症病毒和巴氏杆菌的攻毒产生免疫保护作用，且Vp60-PlpE重组蛋白具有与两种蛋白物理混合物疫苗相同的保护力。
实施例 5 工艺及耗时比较
本专利所提供的嵌合抗原与两种蛋白混合物疫苗虽然具有相似的免疫保护效果，但是制备工艺差别明显。如表1所示，嵌合抗原制备过程不需要经历纯化抗原、内毒素去除、物理混合等步骤，制备时间缩短了50%，也不存在相应步骤的物料、水电消耗，生产成本显著降低，具有很好的应用前景。
表1制备工艺比较
以上显示和描述了本发明的基本原理、主要特征和本发明的优点。本行业的技术人员应该了解，本发明不受上述实施例的限制，上述实施例和说明书中描述的只是说明本发明的原理，在不脱离本发明精神和范围的前提下，本发明还会有各种变化和改进，这些变化和改进都落入要求保护的本发明范围内。本发明要求保护范围由所附的权利要求书及其等效物界定。
序列表
<110> 江苏省农业科学院
<120> 单一位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原及其制备和应用
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Gly Ser Ala Gly Asn Arg Ala Asp Gln Ala Gln Pro Val Gln Leu Pro
1 5 10 15
Gln Ile Val Ile Pro Pro Gln Lys Thr
20 25
<210> 2
<211> 45
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 2
Ser Leu Ala Asn Lys Gln Gln Ile Gln Ile Pro Thr Thr Gln Asn Ser
1 5 10 15
Phe Lys His Ile Glu Glu Glu Lys Met Ile Lys Asp Thr Thr Asn Asp
20 25 30
Trp Phe Lys Ser Cys Val Ser Ser Tyr Gln Asn Thr Cys
35 40 45
<210> 3
<211> 75
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
ggtagcgctg gaaatcgtgc tgaccaagca caaccggttc aactacccca aattgttatc 60
ccacctcaaa aaacg 75
<210> 4
<211> 135
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
tcactagcaa ataaacaaca aattcaaata cctacaactc aaaattcttt taaacatatc 60
gaagaagaga aaatgataaa ggataccaca aacgattggt ttaaaagttg cgtttcctct 120
tatcagaata cttgt 135
<210> 5
<211> 649
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 5
Met Gly Ser Ala Gly Asn Arg Ala Asp Gln Ala Gln Pro Val Gln Leu
1 5 10 15
Pro Gln Ile Val Ile Pro Pro Gln Lys Thr Glu Gly Lys Ala Arg Thr
20 25 30
Ala Pro Gln Gly Glu Ala Ala Gly Thr Ala Thr Thr Ala Ser Val Pro
35 40 45
Gly Thr Thr Thr Asp Gly Met Asp Pro Gly Val Val Ala Ala Thr Ser
50 55 60
Val Val Thr Ala Glu Asn Ser Ser Ala Ser Val Ala Thr Ala Gly Ile
65 70 75 80
Gly Gly Pro Pro Gln Gln Val Asp Gln Gln Glu Thr Trp Arg Thr Asn
85 90 95
Phe Tyr Tyr Asn Asp Val Phe Thr Trp Ser Val Ala Asp Ala Pro Gly
100 105 110
Ser Ile Leu Tyr Thr Val Gln His Ser Pro Gln Asn Asn Pro Phe Thr
115 120 125
Ala Val Leu Ser Gln Met Tyr Ala Gly Trp Ala Gly Gly Met Gln Phe
130 135 140
Arg Phe Ile Val Ala Gly Ser Gly Val Phe Gly Gly Arg Leu Val Ala
145 150 155 160
Ala Val Ile Pro Pro Gly Ile Glu Ile Gly Pro Gly Leu Glu Val Arg
165 170 175
Gln Phe Pro His Val Val Ile Asp Ala Arg Ser Leu Glu Pro Val Thr
180 185 190
Ile Thr Met Pro Asp Leu Arg Pro Asn Met Tyr His Pro Thr Gly Asp
195 200 205
Pro Gly Leu Val Pro Thr Leu Val Leu Ser Val Tyr Asn Asn Leu Ile
210 215 220
Asn Pro Phe Gly Gly Ser Thr Asn Ala Ile Gln Val Thr Val Glu Thr
225 230 235 240
Arg Pro Ser Asp Asp Phe Glu Phe Val Met Ile Arg Ala Pro Ser Ser
245 250 255
Lys Thr Val Asp Ser Ile Ser Pro Ala Gly Leu Leu Thr Thr Pro Val
260 265 270
Leu Thr Gly Val Gly Asn Asp Asn Arg Trp Asn Gly Gln Ile Val Gly
275 280 285
Leu Gln Pro Val Pro Gly Gly Phe Ser Thr Cys Asn Arg His Trp Asn
290 295 300
Leu Asn Gly Ser Thr Tyr Gly Trp Ser Ser Pro Arg Phe Ala Asp Ile
305 310 315 320
Asp His Arg Arg Gly Ser Ala Ser Tyr Ser Gly Asn Asn Ser Thr Asn
325 330 335
Val Leu Gln Phe Trp Tyr Ala Asn Ala Gly Ser Ala Ile Asp Asn Pro
340 345 350
Ile Ser Gln Val Ala Pro Asp Gly Phe Pro Asp Met Ser Phe Val Pro
355 360 365
Phe Asn Ser Pro Asn Ile Pro Thr Ala Gly Trp Val Gly Phe Gly Gly
370 375 380
Ile Trp Asn Ser Asn Asn Gly Ala Pro Ala Ala Thr Thr Val Gln Ala
385 390 395 400
Tyr Glu Leu Gly Phe Ala Thr Gly Ala Pro Asn Ser Leu Gln Pro Thr
405 410 415
Thr Asn Thr Ser Gly Ala Gln Thr Val Ala Lys Ser Ile Tyr Ala Val
420 425 430
Val Thr Gly Thr Asn Gln Asn Pro Thr Gly Leu Phe Val Met Ala Ser
435 440 445
Gly Val Ile Ser Thr Pro Asn Ala Ser Ala Val Thr Tyr Thr Pro Gln
450 455 460
Pro Asp Arg Ile Val Thr Thr Pro Gly Thr Pro Ala Ala Ala Pro Val
465 470 475 480
Gly Lys Asn Thr Pro Ile Met Phe Ala Ser Val Val Arg Arg Thr Gly
485 490 495
Asp Val Asn Ala Ala Ala Gly Ser Thr Asn Gly Thr Gln Tyr Gly Thr
500 505 510
Gly Ser Gln Pro Leu Pro Val Thr Ile Gly Leu Ser Leu Asn Asn Tyr
515 520 525
Ser Ser Ala Leu Val Pro Gly Gln Phe Phe Val Trp Gln Leu Thr Phe
530 535 540
Ala Ser Gly Phe Met Glu Ile Gly Leu Ser Val Asp Gly Tyr Phe Tyr
545 550 555 560
Ala Gly Thr Gly Ala Ser Thr Thr Leu Ile Asp Leu Thr Glu Leu Ile
565 570 575
Asp Val Arg Pro Val Gly Pro Arg Pro Ser Lys Ser Thr Leu Val Phe
580 585 590
Asn Leu Gly Gly Thr Thr Asn Gly Phe Ser Tyr Val Ser Leu Ala Asn
595 600 605
Lys Gln Gln Ile Gln Ile Pro Thr Thr Gln Asn Ser Phe Lys His Ile
610 615 620
Glu Glu Glu Lys Met Ile Lys Asp Thr Thr Asn Asp Trp Phe Lys Ser
625 630 635 640
Cys Val Ser Ser Tyr Gln Asn Thr Cys
645
<210> 6
<211> 1950
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Ala Thr Gly Gly Gly Thr Ala Gly Cys Gly Cys Thr Gly Gly Ala Ala
1 5 10 15
Ala Thr Cys Gly Thr Gly Cys Thr Gly Ala Cys Cys Ala Ala Gly Cys
20 25 30
Ala Cys Ala Ala Cys Cys Gly Gly Thr Thr Cys Ala Ala Cys Thr Ala
35 40 45
Cys Cys Cys Cys Ala Ala Ala Thr Thr Gly Thr Thr Ala Thr Cys Cys
50 55 60
Cys Ala Cys Cys Thr Cys Ala Ala Ala Ala Ala Ala Cys Gly Gly Ala
65 70 75 80
Gly Gly Gly Cys Ala Ala Ala Gly Cys Cys Cys Gly Cys Ala Cys Ala
85 90 95
Gly Cys Gly Cys Cys Gly Cys Ala Ala Gly Gly Cys Gly Ala Ala Gly
100 105 110
Cys Ala Gly Cys Ala Gly Gly Cys Ala Cys Thr Gly Cys Thr Ala Cys
115 120 125
Cys Ala Cys Ala Gly Cys Ala Thr Cys Ala Gly Thr Thr Cys Cys Cys
130 135 140
Gly Gly Ala Ala Cys Cys Ala Cys Gly Ala Cys Cys Gly Ala Cys Gly
145 150 155 160
Gly Cys Ala Thr Gly Gly Ala Thr Cys Cys Thr Gly Gly Thr Gly Thr
165 170 175
Ala Gly Thr Gly Gly Cys Cys Gly Cys Ala Ala Cys Thr Ala Gly Thr
180 185 190
Gly Thr Gly Gly Thr Cys Ala Cys Thr Gly Cys Ala Gly Ala Ala Ala
195 200 205
Ala Thr Thr Cys Ala Thr Cys Cys Gly Cys Ala Thr Cys Gly Gly Thr
210 215 220
Thr Gly Cys Ala Ala Cys Gly Gly Cys Gly Gly Gly Gly Ala Thr Thr
225 230 235 240
Gly Gly Cys Gly Gly Thr Cys Cys Ala Cys Cys Cys Cys Ala Ala Cys
245 250 255
Ala Gly Gly Thr Gly Gly Ala Cys Cys Ala Ala Cys Ala Ala Gly Ala
260 265 270
Ala Ala Cys Ala Thr Gly Gly Ala Gly Gly Ala Cys Ala Ala Ala Cys
275 280 285
Thr Thr Cys Thr Ala Cys Thr Ala Cys Ala Ala Thr Gly Ala Thr Gly
290 295 300
Thr Thr Thr Thr Cys Ala Cys Thr Thr Gly Gly Thr Cys Cys Gly Thr
305 310 315 320
Cys Gly Cys Gly Gly Ala Cys Gly Cys Ala Cys Cys Cys Gly Gly Cys
325 330 335
Ala Gly Cys Ala Thr Thr Cys Thr Cys Thr Ala Cys Ala Cys Thr Gly
340 345 350
Thr Cys Cys Ala Ala Cys Ala Cys Thr Cys Thr Cys Cys Ala Cys Ala
355 360 365
Gly Ala Ala Cys Ala Ala Cys Cys Cys Ala Thr Thr Cys Ala Cys Ala
370 375 380
Gly Cys Thr Gly Thr Ala Cys Thr Gly Ala Gly Cys Cys Ala Gly Ala
385 390 395 400
Thr Gly Thr Ala Cys Gly Cys Thr Gly Gly Cys Thr Gly Gly Gly Cys
405 410 415
Thr Gly Gly Thr Gly Gly Cys Ala Thr Gly Cys Ala Gly Thr Thr Cys
420 425 430
Cys Gly Cys Thr Thr Cys Ala Thr Ala Gly Thr Thr Gly Cys Thr Gly
435 440 445
Gly Ala Thr Cys Ala Gly Gly Thr Gly Thr Gly Thr Thr Thr Gly Gly
450 455 460
Thr Gly Gly Gly Cys Gly Ala Cys Thr Gly Gly Thr Cys Gly Cys Gly
465 470 475 480
Gly Cys Thr Gly Thr Gly Ala Thr Ala Cys Cys Ala Cys Cys Ala Gly
485 490 495
Gly Cys Ala Thr Cys Gly Ala Gly Ala Thr Thr Gly Gly Gly Cys Cys
500 505 510
Ala Gly Gly Gly Thr Thr Gly Gly Ala Gly Gly Thr Cys Ala Gly Gly
515 520 525
Cys Ala Ala Thr Thr Thr Cys Cys Thr Cys Ala Thr Gly Thr Thr Gly
530 535 540
Thr Thr Ala Thr Cys Gly Ala Cys Gly Cys Cys Cys Gly Thr Thr Cys
545 550 555 560
Ala Cys Thr Cys Gly Ala Ala Cys Cys Thr Gly Thr Thr Ala Cys Cys
565 570 575
Ala Thr Cys Ala Cys Cys Ala Thr Gly Cys Cys Ala Gly Ala Cys Thr
580 585 590
Thr Gly Cys Gly Thr Cys Cys Cys Ala Ala Cys Ala Thr Gly Thr Ala
595 600 605
Cys Cys Ala Thr Cys Cys Ala Ala Cys Thr Gly Gly Thr Gly Ala Cys
610 615 620
Cys Cys Thr Gly Gly Cys Cys Thr Thr Gly Thr Cys Cys Cys Cys Ala
625 630 635 640
Cys Ala Cys Thr Ala Gly Thr Cys Cys Thr Thr Ala Gly Thr Gly Thr
645 650 655
Thr Thr Ala Cys Ala Ala Cys Ala Ala Cys Cys Thr Cys Ala Thr Cys
660 665 670
Ala Ala Cys Cys Cys Gly Thr Thr Thr Gly Gly Thr Gly Gly Ala Thr
675 680 685
Cys Cys Ala Cys Cys Ala Ala Cys Gly Cys Ala Ala Thr Cys Cys Ala
690 695 700
Gly Gly Thr Gly Ala Cys Ala Gly Thr Gly Gly Ala Ala Ala Cys Gly
705 710 715 720
Ala Gly Gly Cys Cys Gly Ala Gly Thr Gly Ala Thr Gly Ala Cys Thr
725 730 735
Thr Thr Gly Ala Gly Thr Thr Cys Gly Thr Gly Ala Thr Gly Ala Thr
740 745 750
Thr Ala Gly Ala Gly Cys Cys Cys Cys Cys Thr Cys Cys Ala Gly Cys
755 760 765
Ala Ala Ala Ala Cys Thr Gly Thr Thr Gly Ala Cys Thr Cys Ala Ala
770 775 780
Thr Cys Thr Cys Ala Cys Cys Cys Gly Cys Ala Gly Gly Cys Cys Thr
785 790 795 800
Thr Cys Thr Cys Ala Cys Gly Ala Cys Cys Cys Cys Ala Gly Thr Cys
805 810 815
Cys Thr Cys Ala Cys Thr Gly Gly Thr Gly Thr Thr Gly Gly Cys Ala
820 825 830
Ala Thr Gly Ala Cys Ala Ala Cys Ala Gly Gly Thr Gly Gly Ala Ala
835 840 845
Cys Gly Gly Cys Cys Ala Ala Ala Thr Ala Gly Thr Gly Gly Gly Ala
850 855 860
Cys Thr Gly Cys Ala Ala Cys Cys Ala Gly Thr Ala Cys Cys Thr Gly
865 870 875 880
Gly Gly Gly Gly Gly Thr Thr Thr Thr Cys Cys Ala Cys Gly Thr Gly
885 890 895
Cys Ala Ala Cys Ala Gly Gly Cys Ala Cys Thr Gly Gly Ala Ala Cys
900 905 910
Cys Thr Gly Ala Ala Cys Gly Gly Cys Ala Gly Cys Ala Cys Ala Thr
915 920 925
Ala Thr Gly Gly Cys Thr Gly Gly Thr Cys Ala Ala Gly Cys Cys Cys
930 935 940
Thr Cys Gly Gly Thr Thr Thr Gly Cys Cys Gly Ala Cys Ala Thr Thr
945 950 955 960
Gly Ala Cys Cys Ala Thr Cys Gly Ala Ala Gly Ala Gly Gly Cys Ala
965 970 975
Gly Thr Gly Cys Ala Ala Gly Thr Thr Ala Thr Thr Cys Thr Gly Gly
980 985 990
Gly Ala Ala Cys Ala Ala Cys Thr Cys Cys Ala Cys Cys Ala Ala Cys
995 1000 1005
Gly Thr Gly Cys Thr Thr Cys Ala Gly Thr Thr Thr Thr Gly Gly Thr
1010 1015 1020
Ala Cys Gly Cys Thr Ala Ala Thr Gly Cys Thr Gly Gly Gly Thr Cys
1025 1030 1035 1040
Thr Gly Cys Gly Ala Thr Thr Gly Ala Cys Ala Ala Cys Cys Cys Thr
1045 1050 1055
Ala Thr Cys Thr Cys Cys Cys Ala Gly Gly Thr Thr Gly Cys Ala Cys
1060 1065 1070
Cys Ala Gly Ala Cys Gly Gly Cys Thr Thr Cys Cys Cys Thr Gly Ala
1075 1080 1085
Cys Ala Thr Gly Thr Cys Ala Thr Thr Cys Gly Thr Gly Cys Cys Cys
1090 1095 1100
Thr Thr Thr Ala Ala Cys Ala Gly Cys Cys Cys Cys Ala Ala Cys Ala
1105 1110 1115 1120
Thr Thr Cys Cys Gly Ala Cys Cys Gly Cys Gly Gly Gly Gly Thr Gly
1125 1130 1135
Gly Gly Thr Cys Gly Gly Gly Thr Thr Thr Gly Gly Thr Gly Gly Thr
1140 1145 1150
Ala Thr Thr Thr Gly Gly Ala Ala Cys Ala Gly Thr Ala Ala Cys Ala
1155 1160 1165
Ala Cys Gly Gly Thr Gly Cys Cys Cys Cys Cys Gly Cys Thr Gly Cys
1170 1175 1180
Thr Ala Cys Ala Ala Cys Thr Gly Thr Gly Cys Ala Gly Gly Cys Cys
1185 1190 1195 1200
Thr Ala Thr Gly Ala Gly Thr Thr Ala Gly Gly Thr Thr Thr Thr Gly
1205 1210 1215
Cys Cys Ala Cys Thr Gly Gly Gly Gly Cys Ala Cys Cys Ala Ala Ala
1220 1225 1230
Cys Ala Gly Cys Cys Thr Cys Cys Ala Gly Cys Cys Cys Ala Cys Cys
1235 1240 1245
Ala Cys Cys Ala Ala Cys Ala Cys Thr Thr Cys Ala Gly Gly Thr Gly
1250 1255 1260
Cys Ala Cys Ala Gly Ala Cys Thr Gly Thr Cys Gly Cys Thr Ala Ala
1265 1270 1275 1280
Gly Thr Cys Cys Ala Thr Thr Thr Ala Thr Gly Cys Cys Gly Thr Gly
1285 1290 1295
Gly Thr Ala Ala Cys Cys Gly Gly Cys Ala Cys Ala Ala Ala Cys Cys
1300 1305 1310
Ala Ala Ala Ala Thr Cys Cys Ala Ala Cys Cys Gly Gly Ala Cys Thr
1315 1320 1325
Gly Thr Thr Thr Gly Thr Gly Ala Thr Gly Gly Cys Cys Thr Cys Gly
1330 1335 1340
Gly Gly Thr Gly Thr Thr Ala Thr Cys Thr Cys Cys Ala Cys Gly Cys
1345 1350 1355 1360
Cys Ala Ala Ala Cys Gly Cys Cys Ala Gly Cys Gly Cys Cys Gly Thr
1365 1370 1375
Cys Ala Cys Ala Thr Ala Cys Ala Cys Gly Cys Cys Cys Cys Ala Ala
1380 1385 1390
Cys Cys Ala Gly Ala Cys Ala Gly Ala Ala Thr Thr Gly Thr Gly Ala
1395 1400 1405
Cys Thr Ala Cys Ala Cys Cys Cys Gly Gly Cys Ala Cys Thr Cys Cys
1410 1415 1420
Thr Gly Cys Cys Gly Cys Thr Gly Cys Ala Cys Cys Thr Gly Thr Ala
1425 1430 1435 1440
Gly Gly Thr Ala Ala Gly Ala Ala Cys Ala Cys Ala Cys Cys Cys Ala
1445 1450 1455
Thr Cys Ala Thr Gly Thr Thr Cys Gly Cys Gly Thr Cys Thr Gly Thr
1460 1465 1470
Thr Gly Thr Cys Ala Gly Gly Cys Gly Cys Ala Cys Cys Gly Gly Thr
1475 1480 1485
Gly Ala Cys Gly Thr Cys Ala Ala Cys Gly Cys Cys Gly Cys Ala Gly
1490 1495 1500
Cys Cys Gly Gly Gly Thr Cys Ala Ala Cys Cys Ala Ala Cys Gly Gly
1505 1510 1515 1520
Gly Ala Cys Cys Cys Ala Gly Thr Ala Thr Gly Gly Cys Ala Cys Gly
1525 1530 1535
Gly Gly Cys Thr Cys Cys Cys Ala Ala Cys Cys Ala Cys Thr Gly Cys
1540 1545 1550
Cys Ala Gly Thr Gly Ala Cys Ala Ala Thr Thr Gly Gly Ala Cys Thr
1555 1560 1565
Thr Thr Cys Gly Cys Thr Cys Ala Ala Cys Ala Ala Cys Thr Ala Cys
1570 1575 1580
Thr Cys Gly Thr Cys Ala Gly Cys Ala Cys Thr Cys Gly Thr Gly Cys
1585 1590 1595 1600
Cys Thr Gly Gly Gly Cys Ala Gly Thr Thr Cys Thr Thr Cys Gly Thr
1605 1610 1615
Thr Thr Gly Gly Cys Ala Gly Thr Thr Ala Ala Cys Cys Thr Thr Thr
1620 1625 1630
Gly Cys Ala Thr Cys Thr Gly Gly Thr Thr Thr Cys Ala Thr Gly Gly
1635 1640 1645
Ala Gly Ala Thr Cys Gly Gly Cys Cys Thr Ala Ala Gly Thr Gly Thr
1650 1655 1660
Gly Gly Ala Cys Gly Gly Gly Thr Ala Cys Thr Thr Thr Thr Ala Thr
1665 1670 1675 1680
Gly Cys Ala Gly Gly Ala Ala Cys Ala Gly Gly Ala Gly Cys Cys Thr
1685 1690 1695
Cys Ala Ala Cys Cys Ala Cys Gly Cys Thr Cys Ala Thr Thr Gly Ala
1700 1705 1710
Cys Thr Thr Gly Ala Cys Thr Gly Ala Ala Cys Thr Cys Ala Thr Thr
1715 1720 1725
Gly Ala Cys Gly Thr Ala Cys Gly Cys Cys Cys Cys Gly Thr Gly Gly
1730 1735 1740
Gly Ala Cys Cys Cys Ala Gly Gly Cys Cys Gly Thr Cys Cys Ala Ala
1745 1750 1755 1760
Ala Ala Gly Cys Ala Cys Ala Cys Thr Cys Gly Thr Gly Thr Thr Cys
1765 1770 1775
Ala Ala Cys Cys Thr Gly Gly Gly Gly Gly Gly Cys Ala Cys Ala Ala
1780 1785 1790
Cys Cys Ala Ala Thr Gly Gly Cys Thr Thr Thr Thr Cys Thr Thr Ala
1795 1800 1805
Thr Gly Thr Cys Thr Cys Ala Cys Thr Ala Gly Cys Ala Ala Ala Thr
1810 1815 1820
Ala Ala Ala Cys Ala Ala Cys Ala Ala Ala Thr Thr Cys Ala Ala Ala
1825 1830 1835 1840
Thr Ala Cys Cys Thr Ala Cys Ala Ala Cys Thr Cys Ala Ala Ala Ala
1845 1850 1855
Thr Thr Cys Thr Thr Thr Thr Ala Ala Ala Cys Ala Thr Ala Thr Cys
1860 1865 1870
Gly Ala Ala Gly Ala Ala Gly Ala Gly Ala Ala Ala Ala Thr Gly Ala
1875 1880 1885
Thr Ala Ala Ala Gly Gly Ala Thr Ala Cys Cys Ala Cys Ala Ala Ala
1890 1895 1900
Cys Gly Ala Thr Thr Gly Gly Thr Thr Thr Ala Ala Ala Ala Gly Thr
1905 1910 1915 1920
Thr Gly Cys Gly Thr Thr Thr Cys Cys Thr Cys Thr Thr Ala Thr Cys
1925 1930 1935
Ala Gly Ala Ala Thr Ala Cys Thr Thr Gly Thr Thr Gly Ala
1940 1945 1950
Claims (8)
1.一种双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原，其特征在于，所述重组抗原包括兔出血症病毒VP60蛋白，且其N端和C端分别连接了PlpE up多肽片段和PlpE down多肽片段；其中，PlpE up多肽片段和PlpE down多肽片段源自巴氏杆菌PlpE蛋白抗原表位。
2.根据权利要求1所述一种双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原，其特征在于，所述VP60蛋白N端和C端所嵌入的PlpE抗原表位氨基酸序列分别为SEQ ID NO:1和SEQ ID NO:2。
3.根据权利要求1所述一种双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原，其特征在于，编码所述VP60蛋白N端和C端所嵌入的PlpE抗原表位核苷酸序列分别为SEQ ID NO:3和SEQ ID NO:4。
4.根据权利要求1所述一种双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原，其特征在于，所述双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原的氨基酸序列为SEQ ID NO:5。
5.根据权利要求1所述一种双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原，其特征在于，编码所述双位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原的核苷酸序列为SEQ ID NO:6。
6.一种重组抗原的表达载体，其特征在于，所述载体用于表达双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原，含有权利要求5所述核苷酸序列。
7.一种制备权利要求1所述双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原制备方法，其步骤如下：
(1)通过PCR扩增巴氏杆菌PlpE的第26位到第50位氨基酸所对应的DNA序列、第51位到第95位氨基酸对应的DNA序列;
(2)将步骤(1)得到PCR产物依次分别连接到VP60基因的5’端和3’端上，并将重组基因连接到pFastBac1克隆位点上,得到重组穿梭质粒载体;
(3)将步骤(2)得到的重组穿梭载体转化到DH10Bac宿主菌中，通过该菌的转座作用获得杆状病毒质粒；
（4）将重组杆状病毒质粒转入sf9细胞后获得嵌合了巴氏杆菌PlpE表位的兔出血症病毒VP60重组蛋白抗原。
8.权利要求1所述双位点嵌合了巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原在制备预防兔病毒性出血症和兔巴氏杆菌病疫苗方面的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110553190.XA CN113402615B (zh) | 2021-05-20 | 2021-05-20 | 双位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原及其制备和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110553190.XA CN113402615B (zh) | 2021-05-20 | 2021-05-20 | 双位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原及其制备和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113402615A true CN113402615A (zh) | 2021-09-17 |
| CN113402615B CN113402615B (zh) | 2024-02-23 |
Family
ID=77679060
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110553190.XA Active CN113402615B (zh) | 2021-05-20 | 2021-05-20 | 双位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原及其制备和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113402615B (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115772506A (zh) * | 2022-12-09 | 2023-03-10 | 内蒙古自治区农牧业科学院 | 一种溶血性曼氏杆菌plpe蛋白单克隆抗体及其制备方法和应用 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108904796A (zh) * | 2018-07-12 | 2018-11-30 | 江苏省农业科学院 | 兔出血症病毒杆状病毒载体、多杀性巴氏杆菌病二联灭活疫苗及其制备方法 |
| CN110201153A (zh) * | 2019-06-21 | 2019-09-06 | 齐鲁动物保健品有限公司 | 一种兔病毒性出血症、巴氏杆菌病、波氏杆菌病三联灭活疫苗及其制备方法 |
| CN111643659A (zh) * | 2020-05-15 | 2020-09-11 | 江苏省农业科学院 | 一种兔出血症病毒杆状病毒载体疫苗及其制备方法 |
-
2021
- 2021-05-20 CN CN202110553190.XA patent/CN113402615B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108904796A (zh) * | 2018-07-12 | 2018-11-30 | 江苏省农业科学院 | 兔出血症病毒杆状病毒载体、多杀性巴氏杆菌病二联灭活疫苗及其制备方法 |
| CN110201153A (zh) * | 2019-06-21 | 2019-09-06 | 齐鲁动物保健品有限公司 | 一种兔病毒性出血症、巴氏杆菌病、波氏杆菌病三联灭活疫苗及其制备方法 |
| CN111643659A (zh) * | 2020-05-15 | 2020-09-11 | 江苏省农业科学院 | 一种兔出血症病毒杆状病毒载体疫苗及其制备方法 |
Non-Patent Citations (1)
| Title |
|---|
| 朱伟峰等: "嵌合了巴氏杆菌抗原表位的兔出血症病毒VP60 重组蛋白的构建", 中国动物传染病学报, pages 3 - 5 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115772506A (zh) * | 2022-12-09 | 2023-03-10 | 内蒙古自治区农牧业科学院 | 一种溶血性曼氏杆菌plpe蛋白单克隆抗体及其制备方法和应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113402615B (zh) | 2024-02-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2577280B2 (ja) | 組換えポックスウイルス及び連鎖球菌ｍタンパク質ワクチン | |
| CN109867727B (zh) | 一种flagellin-fiber2融合蛋白、其制备方法和应用 | |
| CN107227311B (zh) | 重组猪细小病毒样粒子及其制备方法和应用 | |
| CN111154778B (zh) | 禽新城疫病毒新型基因工程亚单位疫苗 | |
| CN113603754B (zh) | 一种水禽h5n8亚型流感病毒ha重组蛋白及其制备方法与应用 | |
| CN113402615B (zh) | 双位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原及其制备和应用 | |
| CN113336858B (zh) | 单一位点嵌合巴氏杆菌PlpE抗原表位的兔出血症病毒VP60重组抗原及其制备和应用 | |
| CN110564751A (zh) | 微环dna疫苗设计及应用 | |
| CN109776657B (zh) | 重组诺如病毒vlp颗粒和制备方法及其用途 | |
| CN111607605B (zh) | 一种多价表位和亚单位疫苗的构建方法 | |
| CN110237244B (zh) | 鸭坦布苏病毒基因工程亚单位疫苗及其制备方法和应用 | |
| KR102477253B1 (ko) | 강화된 조류 인플루엔자 바이러스-유사입자를 포함하는 백신 조성물 및 이의 제조 방법 | |
| CN113855795B (zh) | 禽戊型肝炎病毒orf2亚单位疫苗 | |
| CN104292338A (zh) | 含sars病毒n抗原的重组蛋白及展示n蛋白的杆状病毒 | |
| CN114058634B (zh) | 一种鸡滑液囊支原体基因工程亚单位疫苗 | |
| CA2730741C (en) | Self-replicating vector lacking an antibiotic-resistance gene | |
| CN111253494B (zh) | 猪口蹄疫病毒和猪轮状病毒的融合蛋白、类病毒颗粒和疫苗及制备方法 | |
| CN111518222B (zh) | 牛轮状病毒融合蛋白和犊牛腹泻多联疫苗 | |
| CN113248576A (zh) | 一种针对冠状病毒的核酸疫苗及其制备方法 | |
| JP3357071B2 (ja) | 新規な融合タンパク質、その遺伝子、組み換えベクター、及び組み換えウイルスとその利用 | |
| CN111253477A (zh) | 猪圆环病毒3型Cap蛋白、核酸、病毒样颗粒、疫苗及制备方法和应用 | |
| CN112592410B (zh) | 犬腺病毒基因工程亚单位疫苗、其制备方法及应用 | |
| CN117511968B (zh) | 表达SARS-CoV-2的S、E和M蛋白的重组金丝雀痘病毒及其构建方法和应用 | |
| CN114045298B (zh) | 一种鸡滑液囊支原体亚单位疫苗及制备方法及应用 | |
| CN115043947B (zh) | 一种克里米亚-刚果出血热病毒Zera-Gn蛋白纳米颗粒、制备方法及其用途 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |