CN112813080B - 一种人工密码子优化的猪瘟病毒e2蛋白及其制备方法与用途 - Google Patents
一种人工密码子优化的猪瘟病毒e2蛋白及其制备方法与用途 Download PDFInfo
- Publication number
- CN112813080B CN112813080B CN201911122079.4A CN201911122079A CN112813080B CN 112813080 B CN112813080 B CN 112813080B CN 201911122079 A CN201911122079 A CN 201911122079A CN 112813080 B CN112813080 B CN 112813080B
- Authority
- CN
- China
- Prior art keywords
- swine fever
- protein
- fever virus
- leu
- thr
- 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.)
- Active
Links
- 101900173079 Classical swine fever virus Envelope glycoprotein E2 Proteins 0.000 title abstract description 21
- 108020004705 Codon Proteins 0.000 title abstract description 14
- 238000002360 preparation method Methods 0.000 title abstract description 10
- 101710125507 Integrase/recombinase Proteins 0.000 claims abstract description 48
- 241000710777 Classical swine fever virus Species 0.000 claims abstract description 38
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 16
- 239000013604 expression vector Substances 0.000 claims abstract description 6
- 239000002773 nucleotide Substances 0.000 claims abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 4
- 210000004027 cell Anatomy 0.000 claims description 9
- 210000004962 mammalian cell Anatomy 0.000 claims description 9
- 239000013612 plasmid Substances 0.000 claims description 6
- 241000282898 Sus scrofa Species 0.000 abstract description 28
- 206010037660 Pyrexia Diseases 0.000 abstract description 24
- 238000003745 diagnosis Methods 0.000 abstract description 6
- 229940031626 subunit vaccine Drugs 0.000 abstract description 5
- 230000002194 synthesizing effect Effects 0.000 abstract description 2
- 229960005486 vaccine Drugs 0.000 description 15
- 230000014509 gene expression Effects 0.000 description 10
- 238000001514 detection method Methods 0.000 description 8
- 210000002966 serum Anatomy 0.000 description 8
- 238000000034 method Methods 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 241000700605 Viruses Species 0.000 description 6
- 238000006206 glycosylation reaction Methods 0.000 description 6
- 208000001726 Classical Swine Fever Diseases 0.000 description 5
- 230000013595 glycosylation Effects 0.000 description 5
- 101710172711 Structural protein Proteins 0.000 description 4
- 108010047857 aspartylglycine Proteins 0.000 description 4
- LXWYCLOUQZZDBD-LIYNQYRNSA-N csfv Chemical compound C([C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)[C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O)C1=CC=C(O)C=C1 LXWYCLOUQZZDBD-LIYNQYRNSA-N 0.000 description 4
- 230000036039 immunity Effects 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 238000005457 optimization Methods 0.000 description 4
- 230000002265 prevention Effects 0.000 description 4
- 230000001681 protective effect Effects 0.000 description 4
- 108010073969 valyllysine Proteins 0.000 description 4
- 208000035473 Communicable disease Diseases 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 241000282326 Felis catus Species 0.000 description 3
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 3
- 229920001213 Polysorbate 20 Polymers 0.000 description 3
- 150000001413 amino acids Chemical group 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000012228 culture supernatant Substances 0.000 description 3
- 108010009298 lysylglutamic acid Proteins 0.000 description 3
- 239000013600 plasmid vector Substances 0.000 description 3
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 3
- 230000009465 prokaryotic expression Effects 0.000 description 3
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- SQKPKIJVWHAWNF-DCAQKATOSA-N Arg-Asp-Lys Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(O)=O SQKPKIJVWHAWNF-DCAQKATOSA-N 0.000 description 2
- UVTGNSWSRSCPLP-UHFFFAOYSA-N Arg-Tyr Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccc(O)cc1)C(=O)O UVTGNSWSRSCPLP-UHFFFAOYSA-N 0.000 description 2
- 101710117545 C protein Proteins 0.000 description 2
- 206010008631 Cholera Diseases 0.000 description 2
- VPQZSNQICFCCSO-BJDJZHNGSA-N Cys-Leu-Ile Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VPQZSNQICFCCSO-BJDJZHNGSA-N 0.000 description 2
- 241000710781 Flaviviridae Species 0.000 description 2
- FMNHBTKMRFVGRO-FOHZUACHSA-N Gly-Asn-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)CN FMNHBTKMRFVGRO-FOHZUACHSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- UCDWNBFOZCZSNV-AVGNSLFASA-N His-Arg-Met Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(O)=O UCDWNBFOZCZSNV-AVGNSLFASA-N 0.000 description 2
- 108010093488 His-His-His-His-His-His Proteins 0.000 description 2
- BZKDJRSZWLPJNI-SRVKXCTJSA-N His-His-Ser Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(O)=O BZKDJRSZWLPJNI-SRVKXCTJSA-N 0.000 description 2
- KVRKAGGMEWNURO-CIUDSAMLSA-N Leu-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(C)C)N KVRKAGGMEWNURO-CIUDSAMLSA-N 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 2
- 241000711558 Pestivirus type 3 Species 0.000 description 2
- XYHMFGGWNOFUOU-QXEWZRGKSA-N Pro-Ile-Gly Chemical compound OC(=O)CNC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]1CCCN1 XYHMFGGWNOFUOU-QXEWZRGKSA-N 0.000 description 2
- ZVEQWRWMRFIVSD-HRCADAONSA-N Pro-Phe-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N3CCC[C@@H]3C(=O)O ZVEQWRWMRFIVSD-HRCADAONSA-N 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- IXCHOHLPHNGFTJ-YUMQZZPRSA-N Ser-Gly-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CO)N IXCHOHLPHNGFTJ-YUMQZZPRSA-N 0.000 description 2
- HMRAQFJFTOLDKW-GUBZILKMSA-N Ser-His-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(O)=O HMRAQFJFTOLDKW-GUBZILKMSA-N 0.000 description 2
- 241000282887 Suidae Species 0.000 description 2
- 102100021696 Syncytin-1 Human genes 0.000 description 2
- 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 2
- OVBMCNDKCWAXMZ-NAKRPEOUSA-N Val-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](C(C)C)N OVBMCNDKCWAXMZ-NAKRPEOUSA-N 0.000 description 2
- DIOSYUIWOQCXNR-ONGXEEELSA-N Val-Lys-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O DIOSYUIWOQCXNR-ONGXEEELSA-N 0.000 description 2
- GBIUHAYJGWVNLN-UHFFFAOYSA-N Val-Ser-Pro Natural products CC(C)C(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O GBIUHAYJGWVNLN-UHFFFAOYSA-N 0.000 description 2
- BGTDGENDNWGMDQ-KJEVXHAQSA-N Val-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N)O BGTDGENDNWGMDQ-KJEVXHAQSA-N 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 2
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 108010057083 glutamyl-aspartyl-leucine Proteins 0.000 description 2
- 108010082286 glycyl-seryl-alanine Proteins 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 108010015792 glycyllysine Proteins 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 108010028295 histidylhistidine Proteins 0.000 description 2
- 108010034529 leucyl-lysine Proteins 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000003472 neutralizing effect Effects 0.000 description 2
- 108010082795 phenylalanyl-arginyl-arginine Proteins 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 230000000405 serological effect Effects 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 108010060175 trypsinogen activation peptide Proteins 0.000 description 2
- 108010035534 tyrosyl-leucyl-alanine Proteins 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 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 1
- NBTGEURICRTMGL-WHFBIAKZSA-N Ala-Gly-Ser Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O NBTGEURICRTMGL-WHFBIAKZSA-N 0.000 description 1
- HHRAXZAYZFFRAM-CIUDSAMLSA-N Ala-Leu-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O HHRAXZAYZFFRAM-CIUDSAMLSA-N 0.000 description 1
- OYJCVIGKMXUVKB-GARJFASQSA-N Ala-Leu-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N OYJCVIGKMXUVKB-GARJFASQSA-N 0.000 description 1
- DXTYEWAQOXYRHZ-KKXDTOCCSA-N Ala-Phe-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N DXTYEWAQOXYRHZ-KKXDTOCCSA-N 0.000 description 1
- HOVPGJUNRLMIOZ-CIUDSAMLSA-N Ala-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C)N HOVPGJUNRLMIOZ-CIUDSAMLSA-N 0.000 description 1
- REWSWYIDQIELBE-FXQIFTODSA-N Ala-Val-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O REWSWYIDQIELBE-FXQIFTODSA-N 0.000 description 1
- YUIGJDNAGKJLDO-JYJNAYRXSA-N Arg-Arg-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O YUIGJDNAGKJLDO-JYJNAYRXSA-N 0.000 description 1
- KMSHNDWHPWXPEC-BQBZGAKWSA-N Arg-Asp-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O KMSHNDWHPWXPEC-BQBZGAKWSA-N 0.000 description 1
- HKRXJBBCQBAGIM-FXQIFTODSA-N Arg-Asp-Ser Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N)CN=C(N)N HKRXJBBCQBAGIM-FXQIFTODSA-N 0.000 description 1
- LLUGJARLJCGLAR-CYDGBPFRSA-N Arg-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](CCCN=C(N)N)N LLUGJARLJCGLAR-CYDGBPFRSA-N 0.000 description 1
- YBZMTKUDWXZLIX-UWVGGRQHSA-N Arg-Leu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YBZMTKUDWXZLIX-UWVGGRQHSA-N 0.000 description 1
- RIQBRKVTFBWEDY-RHYQMDGZSA-N Arg-Lys-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O RIQBRKVTFBWEDY-RHYQMDGZSA-N 0.000 description 1
- FVBZXNSRIDVYJS-AVGNSLFASA-N Arg-Pro-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCCN=C(N)N FVBZXNSRIDVYJS-AVGNSLFASA-N 0.000 description 1
- DNLQVHBBMPZUGJ-BQBZGAKWSA-N Arg-Ser-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O DNLQVHBBMPZUGJ-BQBZGAKWSA-N 0.000 description 1
- NZQFXJKVNUZYAG-BPUTZDHNSA-N Arg-Trp-Cys Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCCN=C(N)N)N)C(=O)N[C@@H](CS)C(O)=O)=CNC2=C1 NZQFXJKVNUZYAG-BPUTZDHNSA-N 0.000 description 1
- NVPHRWNWTKYIST-BPNCWPANSA-N Arg-Tyr-Ala Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 NVPHRWNWTKYIST-BPNCWPANSA-N 0.000 description 1
- CGWVCWFQGXOUSJ-ULQDDVLXSA-N Arg-Tyr-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(O)=O CGWVCWFQGXOUSJ-ULQDDVLXSA-N 0.000 description 1
- HOIFSHOLNKQCSA-FXQIFTODSA-N Asn-Arg-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O HOIFSHOLNKQCSA-FXQIFTODSA-N 0.000 description 1
- DNYRZPOWBTYFAF-IHRRRGAJSA-N Asn-Arg-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(=O)N)N)O DNYRZPOWBTYFAF-IHRRRGAJSA-N 0.000 description 1
- FTCGGKNCJZOPNB-WHFBIAKZSA-N Asn-Gly-Ser Chemical compound NC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O FTCGGKNCJZOPNB-WHFBIAKZSA-N 0.000 description 1
- XLHLPYFMXGOASD-CIUDSAMLSA-N Asn-His-Asp Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N XLHLPYFMXGOASD-CIUDSAMLSA-N 0.000 description 1
- ULZOQOKFYMXHPZ-AQZXSJQPSA-N Asn-Trp-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ULZOQOKFYMXHPZ-AQZXSJQPSA-N 0.000 description 1
- XEGZSHSPQNDNRH-JRQIVUDYSA-N Asn-Tyr-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XEGZSHSPQNDNRH-JRQIVUDYSA-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
- ZRAOLTNMSCSCLN-ZLUOBGJFSA-N Asp-Cys-Asn Chemical compound C([C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)C(=O)O ZRAOLTNMSCSCLN-ZLUOBGJFSA-N 0.000 description 1
- PJERDVUTUDZPGX-ZKWXMUAHSA-N Asp-Cys-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CS)NC(=O)[C@@H](N)CC(O)=O PJERDVUTUDZPGX-ZKWXMUAHSA-N 0.000 description 1
- IJHUZMGJRGNXIW-CIUDSAMLSA-N Asp-Glu-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O IJHUZMGJRGNXIW-CIUDSAMLSA-N 0.000 description 1
- VILLWIDTHYPSLC-PEFMBERDSA-N Asp-Glu-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VILLWIDTHYPSLC-PEFMBERDSA-N 0.000 description 1
- QCVXMEHGFUMKCO-YUMQZZPRSA-N Asp-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(O)=O QCVXMEHGFUMKCO-YUMQZZPRSA-N 0.000 description 1
- SVABRQFIHCSNCI-FOHZUACHSA-N Asp-Gly-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O SVABRQFIHCSNCI-FOHZUACHSA-N 0.000 description 1
- HKEZZWQWXWGASX-KKUMJFAQSA-N Asp-Leu-Phe Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 HKEZZWQWXWGASX-KKUMJFAQSA-N 0.000 description 1
- WOPJVEMFXYHZEE-SRVKXCTJSA-N Asp-Phe-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O WOPJVEMFXYHZEE-SRVKXCTJSA-N 0.000 description 1
- YIDFBWRHIYOYAA-LKXGYXEUSA-N Asp-Ser-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YIDFBWRHIYOYAA-LKXGYXEUSA-N 0.000 description 1
- XYPJXLLXNSAWHZ-SRVKXCTJSA-N Asp-Ser-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O XYPJXLLXNSAWHZ-SRVKXCTJSA-N 0.000 description 1
- GCACQYDBDHRVGE-LKXGYXEUSA-N Asp-Thr-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@H](O)C)NC(=O)[C@@H](N)CC(O)=O GCACQYDBDHRVGE-LKXGYXEUSA-N 0.000 description 1
- HCOQNGIHSXICCB-IHRRRGAJSA-N Asp-Tyr-Arg Chemical compound N[C@@H](CC(=O)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)O HCOQNGIHSXICCB-IHRRRGAJSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 108090000565 Capsid Proteins Proteins 0.000 description 1
- 102100023321 Ceruloplasmin Human genes 0.000 description 1
- BNRHLRWCERLRTQ-BPUTZDHNSA-N Cys-Arg-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CS)N BNRHLRWCERLRTQ-BPUTZDHNSA-N 0.000 description 1
- PQHYZJPCYRDYNE-QWRGUYRKSA-N Cys-Gly-Phe Chemical compound [H]N[C@@H](CS)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PQHYZJPCYRDYNE-QWRGUYRKSA-N 0.000 description 1
- CNAMJJOZGXPDHW-IHRRRGAJSA-N Cys-Pro-Phe Chemical compound N[C@@H](CS)C(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1ccccc1)C(O)=O CNAMJJOZGXPDHW-IHRRRGAJSA-N 0.000 description 1
- JTEGHEWKBCTIAL-IXOXFDKPSA-N Cys-Thr-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CS)N)O JTEGHEWKBCTIAL-IXOXFDKPSA-N 0.000 description 1
- FCXJJTRGVAZDER-FXQIFTODSA-N Cys-Val-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O FCXJJTRGVAZDER-FXQIFTODSA-N 0.000 description 1
- YQEHNIKPAOPBNH-DCAQKATOSA-N Cys-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CS)N YQEHNIKPAOPBNH-DCAQKATOSA-N 0.000 description 1
- FNXOZWPPOJRBRE-XGEHTFHBSA-N Cys-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CS)N)O FNXOZWPPOJRBRE-XGEHTFHBSA-N 0.000 description 1
- 238000009007 Diagnostic Kit Methods 0.000 description 1
- 101710121417 Envelope glycoprotein Proteins 0.000 description 1
- 101710091045 Envelope protein Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- VZRAXPGTUNDIDK-GUBZILKMSA-N Gln-Leu-Asn Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N VZRAXPGTUNDIDK-GUBZILKMSA-N 0.000 description 1
- UGEZSPWLJABDAR-KKUMJFAQSA-N Gln-Tyr-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CCC(=O)N)N UGEZSPWLJABDAR-KKUMJFAQSA-N 0.000 description 1
- YYOBUPFZLKQUAX-FXQIFTODSA-N Glu-Asn-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O YYOBUPFZLKQUAX-FXQIFTODSA-N 0.000 description 1
- JVSBYEDSSRZQGV-GUBZILKMSA-N Glu-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCC(O)=O JVSBYEDSSRZQGV-GUBZILKMSA-N 0.000 description 1
- CYHBMLHCQXXCCT-AVGNSLFASA-N Glu-Asp-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CYHBMLHCQXXCCT-AVGNSLFASA-N 0.000 description 1
- UENPHLAAKDPZQY-XKBZYTNZSA-N Glu-Cys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CCC(=O)O)N)O UENPHLAAKDPZQY-XKBZYTNZSA-N 0.000 description 1
- KASDBWKLWJKTLJ-GUBZILKMSA-N Glu-Glu-Met Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O KASDBWKLWJKTLJ-GUBZILKMSA-N 0.000 description 1
- INGJLBQKTRJLFO-UKJIMTQDSA-N Glu-Ile-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCC(O)=O INGJLBQKTRJLFO-UKJIMTQDSA-N 0.000 description 1
- QMOSCLNJVKSHHU-YUMQZZPRSA-N Glu-Met-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O QMOSCLNJVKSHHU-YUMQZZPRSA-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
- GPSHCSTUYOQPAI-JHEQGTHGSA-N Glu-Thr-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O GPSHCSTUYOQPAI-JHEQGTHGSA-N 0.000 description 1
- UCZXXMREFIETQW-AVGNSLFASA-N Glu-Tyr-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O UCZXXMREFIETQW-AVGNSLFASA-N 0.000 description 1
- QGAJQIGFFIQJJK-IHRRRGAJSA-N Glu-Tyr-Gln Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O QGAJQIGFFIQJJK-IHRRRGAJSA-N 0.000 description 1
- SOYWRINXUSUWEQ-DLOVCJGASA-N Glu-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCC(O)=O SOYWRINXUSUWEQ-DLOVCJGASA-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
- FUTAPPOITCCWTH-WHFBIAKZSA-N Gly-Asp-Asp Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O FUTAPPOITCCWTH-WHFBIAKZSA-N 0.000 description 1
- BEQGFMIBZFNROK-JGVFFNPUSA-N Gly-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)CN)C(=O)O BEQGFMIBZFNROK-JGVFFNPUSA-N 0.000 description 1
- KMSGYZQRXPUKGI-BYPYZUCNSA-N Gly-Gly-Asn Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC(N)=O KMSGYZQRXPUKGI-BYPYZUCNSA-N 0.000 description 1
- OLPPXYMMIARYAL-QMMMGPOBSA-N Gly-Gly-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)CNC(=O)CN OLPPXYMMIARYAL-QMMMGPOBSA-N 0.000 description 1
- CQIIXEHDSZUSAG-QWRGUYRKSA-N Gly-His-His Chemical compound C([C@H](NC(=O)CN)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 CQIIXEHDSZUSAG-QWRGUYRKSA-N 0.000 description 1
- HHSOPSCKAZKQHQ-PEXQALLHSA-N Gly-His-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)CN HHSOPSCKAZKQHQ-PEXQALLHSA-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
- LHYJCVCQPWRMKZ-WEDXCCLWSA-N Gly-Leu-Thr Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LHYJCVCQPWRMKZ-WEDXCCLWSA-N 0.000 description 1
- VLIJYPMATZSOLL-YUMQZZPRSA-N Gly-Lys-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)CN VLIJYPMATZSOLL-YUMQZZPRSA-N 0.000 description 1
- GAFKBWKVXNERFA-QWRGUYRKSA-N Gly-Phe-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 GAFKBWKVXNERFA-QWRGUYRKSA-N 0.000 description 1
- BMWFDYIYBAFROD-WPRPVWTQSA-N Gly-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN BMWFDYIYBAFROD-WPRPVWTQSA-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
- JSLVAHYTAJJEQH-QWRGUYRKSA-N Gly-Ser-Phe Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JSLVAHYTAJJEQH-QWRGUYRKSA-N 0.000 description 1
- FKESCSGWBPUTPN-FOHZUACHSA-N Gly-Thr-Asn Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O FKESCSGWBPUTPN-FOHZUACHSA-N 0.000 description 1
- CUVBTVWFVIIDOC-YEPSODPASA-N Gly-Thr-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)CN CUVBTVWFVIIDOC-YEPSODPASA-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
- DZMVESFTHXSSPZ-XVYDVKMFSA-N His-Ala-Ser Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DZMVESFTHXSSPZ-XVYDVKMFSA-N 0.000 description 1
- UOAVQQRILDGZEN-SRVKXCTJSA-N His-Asp-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O UOAVQQRILDGZEN-SRVKXCTJSA-N 0.000 description 1
- UMBKDWGQESDCTO-KKUMJFAQSA-N His-Lys-Lys Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O UMBKDWGQESDCTO-KKUMJFAQSA-N 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- HVWXAQVMRBKKFE-UGYAYLCHSA-N Ile-Asp-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N HVWXAQVMRBKKFE-UGYAYLCHSA-N 0.000 description 1
- QRTVJGKXFSYJGW-KBIXCLLPSA-N Ile-Glu-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CS)C(=O)O)N QRTVJGKXFSYJGW-KBIXCLLPSA-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
- RWYCOSAAAJBJQL-KCTSRDHCSA-N Ile-Gly-Trp Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N RWYCOSAAAJBJQL-KCTSRDHCSA-N 0.000 description 1
- KLBVGHCGHUNHEA-BJDJZHNGSA-N Ile-Leu-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)O)N KLBVGHCGHUNHEA-BJDJZHNGSA-N 0.000 description 1
- PXKACEXYLPBMAD-JBDRJPRFSA-N Ile-Ser-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PXKACEXYLPBMAD-JBDRJPRFSA-N 0.000 description 1
- JZBVBOKASHNXAD-NAKRPEOUSA-N Ile-Val-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N JZBVBOKASHNXAD-NAKRPEOUSA-N 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 1
- KFKWRHQBZQICHA-STQMWFEESA-N L-leucyl-L-phenylalanine Natural products CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KFKWRHQBZQICHA-STQMWFEESA-N 0.000 description 1
- CZCSUZMIRKFFFA-CIUDSAMLSA-N Leu-Ala-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O CZCSUZMIRKFFFA-CIUDSAMLSA-N 0.000 description 1
- BQSLGJHIAGOZCD-CIUDSAMLSA-N Leu-Ala-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O BQSLGJHIAGOZCD-CIUDSAMLSA-N 0.000 description 1
- STAVRDQLZOTNKJ-RHYQMDGZSA-N Leu-Arg-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O STAVRDQLZOTNKJ-RHYQMDGZSA-N 0.000 description 1
- VCSBGUACOYUIGD-CIUDSAMLSA-N Leu-Asn-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O VCSBGUACOYUIGD-CIUDSAMLSA-N 0.000 description 1
- TWQIYNGNYNJUFM-NHCYSSNCSA-N Leu-Asn-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O TWQIYNGNYNJUFM-NHCYSSNCSA-N 0.000 description 1
- HUEBCHPSXSQUGN-GARJFASQSA-N Leu-Cys-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)N1CCC[C@@H]1C(=O)O)N HUEBCHPSXSQUGN-GARJFASQSA-N 0.000 description 1
- RVVBWTWPNFDYBE-SRVKXCTJSA-N Leu-Glu-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RVVBWTWPNFDYBE-SRVKXCTJSA-N 0.000 description 1
- VWHGTYCRDRBSFI-ZETCQYMHSA-N Leu-Gly-Gly Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)NCC(O)=O VWHGTYCRDRBSFI-ZETCQYMHSA-N 0.000 description 1
- YFBBUHJJUXXZOF-UWVGGRQHSA-N Leu-Gly-Pro Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O YFBBUHJJUXXZOF-UWVGGRQHSA-N 0.000 description 1
- IAJFFZORSWOZPQ-SRVKXCTJSA-N Leu-Leu-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IAJFFZORSWOZPQ-SRVKXCTJSA-N 0.000 description 1
- UBZGNBKMIJHOHL-BZSNNMDCSA-N Leu-Leu-Phe Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C([O-])=O)CC1=CC=CC=C1 UBZGNBKMIJHOHL-BZSNNMDCSA-N 0.000 description 1
- LVTJJOJKDCVZGP-QWRGUYRKSA-N Leu-Lys-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O LVTJJOJKDCVZGP-QWRGUYRKSA-N 0.000 description 1
- ZAVCJRJOQKIOJW-KKUMJFAQSA-N Leu-Phe-Asp Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(O)=O)C(O)=O)CC1=CC=CC=C1 ZAVCJRJOQKIOJW-KKUMJFAQSA-N 0.000 description 1
- MUCIDQMDOYQYBR-IHRRRGAJSA-N Leu-Pro-His Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N MUCIDQMDOYQYBR-IHRRRGAJSA-N 0.000 description 1
- PWPBLZXWFXJFHE-RHYQMDGZSA-N Leu-Pro-Thr Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O PWPBLZXWFXJFHE-RHYQMDGZSA-N 0.000 description 1
- GZRABTMNWJXFMH-UVOCVTCTSA-N Leu-Thr-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZRABTMNWJXFMH-UVOCVTCTSA-N 0.000 description 1
- TUIOUEWKFFVNLH-DCAQKATOSA-N Leu-Val-Cys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CS)C(O)=O TUIOUEWKFFVNLH-DCAQKATOSA-N 0.000 description 1
- 239000006142 Luria-Bertani Agar Substances 0.000 description 1
- UWKNTTJNVSYXPC-CIUDSAMLSA-N Lys-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN UWKNTTJNVSYXPC-CIUDSAMLSA-N 0.000 description 1
- DNEJSAIMVANNPA-DCAQKATOSA-N Lys-Asn-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O DNEJSAIMVANNPA-DCAQKATOSA-N 0.000 description 1
- SFQPJNQDUUYCLA-BJDJZHNGSA-N Lys-Cys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CCCCN)N SFQPJNQDUUYCLA-BJDJZHNGSA-N 0.000 description 1
- GUYHHBZCBQZLFW-GUBZILKMSA-N Lys-Gln-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CS)C(=O)O)N GUYHHBZCBQZLFW-GUBZILKMSA-N 0.000 description 1
- ODUQLUADRKMHOZ-JYJNAYRXSA-N Lys-Glu-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCCCN)N)O ODUQLUADRKMHOZ-JYJNAYRXSA-N 0.000 description 1
- LCMWVZLBCUVDAZ-IUCAKERBSA-N Lys-Gly-Glu Chemical compound [NH3+]CCCC[C@H]([NH3+])C(=O)NCC(=O)N[C@H](C([O-])=O)CCC([O-])=O LCMWVZLBCUVDAZ-IUCAKERBSA-N 0.000 description 1
- NKKFVJRLCCUJNA-QWRGUYRKSA-N Lys-Gly-Lys Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN NKKFVJRLCCUJNA-QWRGUYRKSA-N 0.000 description 1
- SKRGVGLIRUGANF-AVGNSLFASA-N Lys-Leu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SKRGVGLIRUGANF-AVGNSLFASA-N 0.000 description 1
- XOQMURBBIXRRCR-SRVKXCTJSA-N Lys-Lys-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCCN XOQMURBBIXRRCR-SRVKXCTJSA-N 0.000 description 1
- RPWTZTBIFGENIA-VOAKCMCISA-N Lys-Thr-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O RPWTZTBIFGENIA-VOAKCMCISA-N 0.000 description 1
- YCJCEMKOZOYBEF-OEAJRASXSA-N Lys-Thr-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O YCJCEMKOZOYBEF-OEAJRASXSA-N 0.000 description 1
- RMOKGALPSPOYKE-KATARQTJSA-N Lys-Thr-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O RMOKGALPSPOYKE-KATARQTJSA-N 0.000 description 1
- PELXPRPDQRFBGQ-KKUMJFAQSA-N Lys-Tyr-Asn Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCCN)N)O PELXPRPDQRFBGQ-KKUMJFAQSA-N 0.000 description 1
- RIPJMCFGQHGHNP-RHYQMDGZSA-N Lys-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCCCN)N)O RIPJMCFGQHGHNP-RHYQMDGZSA-N 0.000 description 1
- ZGVYWHODYWRPLK-GUBZILKMSA-N Met-Pro-Cys Chemical compound CSCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CS)C(O)=O ZGVYWHODYWRPLK-GUBZILKMSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 1
- 241000710778 Pestivirus Species 0.000 description 1
- JEGFCFLCRSJCMA-IHRRRGAJSA-N Phe-Arg-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N JEGFCFLCRSJCMA-IHRRRGAJSA-N 0.000 description 1
- CSYVXYQDIVCQNU-QWRGUYRKSA-N Phe-Asp-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O CSYVXYQDIVCQNU-QWRGUYRKSA-N 0.000 description 1
- SWZKMTDPQXLQRD-XVSYOHENSA-N Phe-Asp-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SWZKMTDPQXLQRD-XVSYOHENSA-N 0.000 description 1
- IILUKIJNFMUBNF-IHRRRGAJSA-N Phe-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O IILUKIJNFMUBNF-IHRRRGAJSA-N 0.000 description 1
- KJJROSNFBRWPHS-JYJNAYRXSA-N Phe-Glu-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O KJJROSNFBRWPHS-JYJNAYRXSA-N 0.000 description 1
- CVAUVSOFHJKCHN-BZSNNMDCSA-N Phe-Tyr-Cys Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CS)C(O)=O)C1=CC=CC=C1 CVAUVSOFHJKCHN-BZSNNMDCSA-N 0.000 description 1
- ZYNBEWGJFXTBDU-ACRUOGEOSA-N Phe-Tyr-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CC2=CC=CC=C2)N ZYNBEWGJFXTBDU-ACRUOGEOSA-N 0.000 description 1
- SGCZFWSQERRKBD-BQBZGAKWSA-N Pro-Asp-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 SGCZFWSQERRKBD-BQBZGAKWSA-N 0.000 description 1
- XFFIGWGYMUFCCQ-ULQDDVLXSA-N Pro-His-Tyr Chemical compound C1=CC(O)=CC=C1C[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)[C@H]1[NH2+]CCC1)CC1=CN=CN1 XFFIGWGYMUFCCQ-ULQDDVLXSA-N 0.000 description 1
- XQPHBAKJJJZOBX-SRVKXCTJSA-N Pro-Lys-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O XQPHBAKJJJZOBX-SRVKXCTJSA-N 0.000 description 1
- ANESFYPBAJPYNJ-SDDRHHMPSA-N Pro-Met-Pro Chemical compound CSCC[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2 ANESFYPBAJPYNJ-SDDRHHMPSA-N 0.000 description 1
- KWMZPPWYBVZIER-XGEHTFHBSA-N Pro-Ser-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KWMZPPWYBVZIER-XGEHTFHBSA-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
- WWXNZNWZNZPDIF-SRVKXCTJSA-N Pro-Val-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 WWXNZNWZNZPDIF-SRVKXCTJSA-N 0.000 description 1
- FHJQROWZEJFZPO-SRVKXCTJSA-N Pro-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 FHJQROWZEJFZPO-SRVKXCTJSA-N 0.000 description 1
- 101710194807 Protective antigen Proteins 0.000 description 1
- 101800004937 Protein C Proteins 0.000 description 1
- 102000017975 Protein C Human genes 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 101710188315 Protein X Proteins 0.000 description 1
- 101800001700 Saposin-D Proteins 0.000 description 1
- BTKUIVBNGBFTTP-WHFBIAKZSA-N Ser-Ala-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)NCC(O)=O BTKUIVBNGBFTTP-WHFBIAKZSA-N 0.000 description 1
- FTVRVZNYIYWJGB-ACZMJKKPSA-N Ser-Asp-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O FTVRVZNYIYWJGB-ACZMJKKPSA-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
- FLONGDPORFIVQW-XGEHTFHBSA-N Ser-Pro-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO FLONGDPORFIVQW-XGEHTFHBSA-N 0.000 description 1
- CKDXFSPMIDSMGV-GUBZILKMSA-N Ser-Pro-Val Chemical compound [H]N[C@@H](CO)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O CKDXFSPMIDSMGV-GUBZILKMSA-N 0.000 description 1
- 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 1
- PYTKULIABVRXSC-BWBBJGPYSA-N Ser-Ser-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PYTKULIABVRXSC-BWBBJGPYSA-N 0.000 description 1
- KKKVOZNCLALMPV-XKBZYTNZSA-N Ser-Thr-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O KKKVOZNCLALMPV-XKBZYTNZSA-N 0.000 description 1
- SIEBDTCABMZCLF-XGEHTFHBSA-N Ser-Val-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SIEBDTCABMZCLF-XGEHTFHBSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- XSLXHSYIVPGEER-KZVJFYERSA-N Thr-Ala-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O XSLXHSYIVPGEER-KZVJFYERSA-N 0.000 description 1
- LXWZOMSOUAMOIA-JIOCBJNQSA-N Thr-Asn-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N)O LXWZOMSOUAMOIA-JIOCBJNQSA-N 0.000 description 1
- OJRNZRROAIAHDL-LKXGYXEUSA-N Thr-Asn-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O OJRNZRROAIAHDL-LKXGYXEUSA-N 0.000 description 1
- NLJKZUGAIIRWJN-LKXGYXEUSA-N Thr-Asp-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CS)C(=O)O)N)O NLJKZUGAIIRWJN-LKXGYXEUSA-N 0.000 description 1
- YBXMGKCLOPDEKA-NUMRIWBASA-N Thr-Asp-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O YBXMGKCLOPDEKA-NUMRIWBASA-N 0.000 description 1
- JQAWYCUUFIMTHE-WLTAIBSBSA-N Thr-Gly-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JQAWYCUUFIMTHE-WLTAIBSBSA-N 0.000 description 1
- JKGGPMOUIAAJAA-YEPSODPASA-N Thr-Gly-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O JKGGPMOUIAAJAA-YEPSODPASA-N 0.000 description 1
- AMXMBCAXAZUCFA-RHYQMDGZSA-N Thr-Leu-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AMXMBCAXAZUCFA-RHYQMDGZSA-N 0.000 description 1
- MEJHFIOYJHTWMK-VOAKCMCISA-N Thr-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)[C@@H](C)O MEJHFIOYJHTWMK-VOAKCMCISA-N 0.000 description 1
- MECLEFZMPPOEAC-VOAKCMCISA-N Thr-Leu-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MECLEFZMPPOEAC-VOAKCMCISA-N 0.000 description 1
- WRQLCVIALDUQEQ-UNQGMJICSA-N Thr-Phe-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WRQLCVIALDUQEQ-UNQGMJICSA-N 0.000 description 1
- KZURUCDWKDEAFZ-XVSYOHENSA-N Thr-Phe-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O KZURUCDWKDEAFZ-XVSYOHENSA-N 0.000 description 1
- NZRUWPIYECBYRK-HTUGSXCWSA-N Thr-Phe-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O NZRUWPIYECBYRK-HTUGSXCWSA-N 0.000 description 1
- IEZVHOULSUULHD-XGEHTFHBSA-N Thr-Ser-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O IEZVHOULSUULHD-XGEHTFHBSA-N 0.000 description 1
- 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 1
- CSZFFQBUTMGHAH-UAXMHLISSA-N Thr-Thr-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N)O CSZFFQBUTMGHAH-UAXMHLISSA-N 0.000 description 1
- LECUEEHKUFYOOV-ZJDVBMNYSA-N Thr-Thr-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](N)[C@@H](C)O LECUEEHKUFYOOV-ZJDVBMNYSA-N 0.000 description 1
- MYNYCUXMIIWUNW-IEGACIPQSA-N Thr-Trp-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MYNYCUXMIIWUNW-IEGACIPQSA-N 0.000 description 1
- PWONLXBUSVIZPH-RHYQMDGZSA-N Thr-Val-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N)O PWONLXBUSVIZPH-RHYQMDGZSA-N 0.000 description 1
- NJNCVQYFNKZMAH-JYBASQMISA-N Trp-Thr-Cys Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CS)C(O)=O)=CNC2=C1 NJNCVQYFNKZMAH-JYBASQMISA-N 0.000 description 1
- NSOMQRHZMJMZIE-GVARAGBVSA-N Tyr-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NSOMQRHZMJMZIE-GVARAGBVSA-N 0.000 description 1
- KDGFPPHLXCEQRN-STECZYCISA-N Tyr-Arg-Ile Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KDGFPPHLXCEQRN-STECZYCISA-N 0.000 description 1
- ZNFPUOSTMUMUDR-JRQIVUDYSA-N Tyr-Asn-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZNFPUOSTMUMUDR-JRQIVUDYSA-N 0.000 description 1
- FFCRCJZJARTYCG-KKUMJFAQSA-N Tyr-Cys-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCCN)C(=O)O)N)O FFCRCJZJARTYCG-KKUMJFAQSA-N 0.000 description 1
- FJKXUIJOMUWCDD-FHWLQOOXSA-N Tyr-Gln-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N)O FJKXUIJOMUWCDD-FHWLQOOXSA-N 0.000 description 1
- LHTGRUZSZOIAKM-SOUVJXGZSA-N Tyr-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)C(=O)O LHTGRUZSZOIAKM-SOUVJXGZSA-N 0.000 description 1
- OFHKXNKJXURPSY-ULQDDVLXSA-N Tyr-Met-Leu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O OFHKXNKJXURPSY-ULQDDVLXSA-N 0.000 description 1
- FDKDGFGTHGJKNV-FHWLQOOXSA-N Tyr-Phe-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N FDKDGFGTHGJKNV-FHWLQOOXSA-N 0.000 description 1
- SOEGLGLDSUHWTI-STECZYCISA-N Tyr-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=C(O)C=C1 SOEGLGLDSUHWTI-STECZYCISA-N 0.000 description 1
- CLEGSEJVGBYZBJ-MEYUZBJRSA-N Tyr-Thr-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H]([C@H](O)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 CLEGSEJVGBYZBJ-MEYUZBJRSA-N 0.000 description 1
- JQOMHZMWQHXALX-FHWLQOOXSA-N Tyr-Tyr-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JQOMHZMWQHXALX-FHWLQOOXSA-N 0.000 description 1
- YODDULVCGFQRFZ-ZKWXMUAHSA-N Val-Asp-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O YODDULVCGFQRFZ-ZKWXMUAHSA-N 0.000 description 1
- DLYOEFGPYTZVSP-AEJSXWLSSA-N Val-Cys-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)N1CCC[C@@H]1C(=O)O)N DLYOEFGPYTZVSP-AEJSXWLSSA-N 0.000 description 1
- CVIXTAITYJQMPE-LAEOZQHASA-N Val-Glu-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CVIXTAITYJQMPE-LAEOZQHASA-N 0.000 description 1
- FEFZWCSXEMVSPO-LSJOCFKGSA-N Val-His-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](C)C(O)=O FEFZWCSXEMVSPO-LSJOCFKGSA-N 0.000 description 1
- RWOGENDAOGMHLX-DCAQKATOSA-N Val-Lys-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C(C)C)N RWOGENDAOGMHLX-DCAQKATOSA-N 0.000 description 1
- WBAJDGWKRIHOAC-GVXVVHGQSA-N Val-Lys-Gln Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O WBAJDGWKRIHOAC-GVXVVHGQSA-N 0.000 description 1
- VPGCVZRRBYOGCD-AVGNSLFASA-N Val-Lys-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O VPGCVZRRBYOGCD-AVGNSLFASA-N 0.000 description 1
- AJNUKMZFHXUBMK-GUBZILKMSA-N Val-Ser-Arg Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N AJNUKMZFHXUBMK-GUBZILKMSA-N 0.000 description 1
- CEKSLIVSNNGOKH-KZVJFYERSA-N Val-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](C(C)C)N)O CEKSLIVSNNGOKH-KZVJFYERSA-N 0.000 description 1
- PQSNETRGCRUOGP-KKHAAJSZSA-N Val-Thr-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(N)=O PQSNETRGCRUOGP-KKHAAJSZSA-N 0.000 description 1
- AOILQMZPNLUXCM-AVGNSLFASA-N Val-Val-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN AOILQMZPNLUXCM-AVGNSLFASA-N 0.000 description 1
- LLJLBRRXKZTTRD-GUBZILKMSA-N Val-Val-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N LLJLBRRXKZTTRD-GUBZILKMSA-N 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 description 1
- 108010068380 arginylarginine Proteins 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 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
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 108010016616 cysteinylglycine Proteins 0.000 description 1
- 108010060199 cysteinylproline Proteins 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 1
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 1
- 108010026364 glycyl-glycyl-leucine Proteins 0.000 description 1
- 108010078326 glycyl-glycyl-valine Proteins 0.000 description 1
- 108010084389 glycyltryptophan Proteins 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 108010078274 isoleucylvaline Proteins 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 230000021633 leukocyte mediated immunity Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 1
- 108010005942 methionylglycine Proteins 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 108010004914 prolylarginine Proteins 0.000 description 1
- 108010053725 prolylvaline Proteins 0.000 description 1
- 229960000856 protein c Drugs 0.000 description 1
- 208000009305 pseudorabies Diseases 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 230000010473 stable expression Effects 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 108010020532 tyrosyl-proline Proteins 0.000 description 1
- 108010078580 tyrosylleucine Proteins 0.000 description 1
- 108010003137 tyrosyltyrosine Proteins 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 230000001018 virulence Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
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
- A61K39/12—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
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—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
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
-
- 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
- C12N2770/24011—Flaviviridae
- C12N2770/24022—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
- C12N2770/24011—Flaviviridae
- C12N2770/24034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/18—Togaviridae; Flaviviridae
- G01N2333/183—Flaviviridae, e.g. pestivirus, mucosal disease virus, bovine viral diarrhoea virus, classical swine fever virus (hog cholera virus) or border disease virus
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Virology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- General Engineering & Computer Science (AREA)
- Hematology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Animal Behavior & Ethology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Urology & Nephrology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Tropical Medicine & Parasitology (AREA)
- Cell Biology (AREA)
- Plant Pathology (AREA)
- Gastroenterology & Hepatology (AREA)
- Food Science & Technology (AREA)
Abstract
本发明属于生物技术领域，涉及一种人工密码子优化的猪瘟病毒E2蛋白制备方法与用途。本发明通过优化与合成可猪瘟病毒E2蛋白编码基因，利用人工合成核苷酸序列构建表达载体。获得的优化猪瘟病毒E2蛋白可用于制备猪瘟亚单位疫苗及制备猪瘟诊断试剂盒。
Description
技术领域
本发明属生物技术领域，涉及一种人工密码子优化的猪瘟病毒E2蛋白及其制备方法与用途。本发明通过优化与合成可猪瘟病毒E2蛋白编码基因，利用人工合成核苷酸序列构建表达载体。获得的优化猪瘟病毒E2蛋白可用于制备猪瘟亚单位疫苗及制备猪瘟诊断试剂盒。
背景技术
据资料记载，猪瘟(Classical swine fever，CSF)又称猪霍乱(Hog cholera)，是由猪瘟病毒(Classical swine fever virus，CSFV)引起的一种急性、高热、接触性传染病，是严重威胁猪健康的重要传染病之一。据记载，猪瘟最早流行在19世纪的美国，随后迅速在世界范围内传播，给世界养猪业造成了严重的经济损失。1984年猪瘟被世界卫生组织(OIE)列为A类传染病，在中国也被列为一类传染病。面对严重危害养猪业的猪瘟疫情，世界各国纷纷不断完善有效的诊断方法和防控技术，猪瘟得以控制。在欧洲的一些国家，经过严格的控制与预防，猪瘟疫情已经达到完全消灭的态势。但是近些年来，猪瘟由早期的急性感染转变为慢性或非典型的症状呈现，这给猪瘟的防治带来极大的挑战。为应对猪瘟疫情越来越严峻的考验，针对猪瘟疫苗的研发及血清抗体诊断方法的建立已引起本领域研究人员的关注，这对世界范围内猪瘟疾病的诊断与防控具有重要意义。
CSFV属于病毒界黄病毒科(Flaviviridae)瘟病毒属(Pestivims)，是单股正链RNA病毒，其基因组编码4个结构蛋白和8个非结构蛋白，其中，4个结构蛋白分别为C、E0、E1和E2，其中C蛋白为衣壳蛋白，其余为囊膜糖蛋白；研究显示，所述囊膜蛋白E0、E1和E2在病毒的脱衣壳与入核阶段起到重要作用，C蛋白在维持病毒核酸稳定起到关键作用。
研究显示，结构蛋白在诱导宿主产生保护性免疫中也起到重要作用，C蛋白可以激活宿主的细胞介导的免疫反应，蛋白不能诱导机体产生中和性抗体，E0蛋白与病毒的毒力相关，其抗体可以阻断病毒对细胞的感染，E2蛋白是CSFV主要保护性抗原蛋白，能激活宿主产生保护性免疫，诱导产生抗CSFV中和抗体，因此，E2蛋白一直作为研制CSFV新型疫苗和血清抗体检测的首选靶蛋白。
研究公开了E2蛋白上分布着5个N-糖基化位点，这些糖基化位点在激活宿主保护性免疫中起到重要的作用；。此外，E2能够形成同源二聚体或者与E1形成异源二聚体，其空间结构包含3个疏水区和3个N端链内二硫键；因此，E2蛋白正确糖基化不仅对于E2蛋白结构的稳定起到关键作用，也对激活宿主免疫功能起到重要作用。目前，重组E2蛋白的制备多采用原核表达系统，虽然原核表达系统操作简单，产量高，但由于细菌不能进行糖基化修饰，原核表达系统制备的重组的E2蛋白不能激活宿主的保护性免疫。有研究利用杆状病毒在昆虫细胞表达系统中可以制备重组E2蛋白，昆虫表达系统虽然可以进行蛋白的糖基化修饰，但由于猪病病毒是感染哺乳细胞，E2的糖基化修饰与昆虫细胞的糖基化修饰具有一定的差别，会导致E2蛋白结构和功能与天然蛋白具有一定的差异。有报道关于哺乳细胞表达系统用于猪瘟E2蛋白制备，但由于产量低，制备成本高，难以用于实际生产。
基于现有技术的现状，本申请的发明人拟提供制备高产量的糖基化猪瘟病毒E2蛋白，不仅可以用于猪瘟的诊断，也可用于猪瘟亚单位疫苗的开发；具体涉及一种人工密码子优化的猪瘟病毒E2蛋白制备方法与用途
发明内容
本发明的目的在于基于现有技术的现状，提供制备高产量的糖基化猪瘟病毒E2蛋白及其方法，具体涉及一种人工密码子优化的猪瘟病毒E2蛋白制备方法与用途，制备的猪瘟病毒E2蛋白不仅可以用于猪瘟的诊断，也可用于猪瘟亚单位疫苗的开发。
本发明提供了一种优化的猪瘟病毒E2蛋白及其编码基因，产生本发明的基因序列的方法不受特别的限制。
本发明中，编码所述的优化的猪瘟病毒E2蛋白编码核苷酸序列为SEQ1，其氨基酸序列为SEQ2。
含有编码本发明的优化的猪瘟病毒E2蛋白及其编码基因的质粒载体构成本发明的一个部分。
本发明提供了一种质粒载体，含有编码所述的优化的猪瘟病毒E2蛋白的DNA，它能插入编码本发明的优化的猪瘟病毒E2蛋白的编码基因；本发明的质粒不受特别限制，优选pcDNA3.4。
表达本发明的含有优化的猪瘟病毒E2蛋白编码基因的宿主细胞构成本发明的一个部分。
本发明提供了一种中华仓鼠卵巢细胞CHO为宿主细胞，它能高效稳定表达本发明优化的猪瘟病毒E2蛋白。本发明的宿主细胞不受特别的限制。
在本发明的实施中，筛选能表达优化的猪瘟病毒E2蛋白的质粒载体的方法不受特别的限制，可通过例如免疫印迹、ELISA等进行选择。
本发明的实施中，纯化优化的猪瘟病毒E2蛋白的方法不受特别的限制，但首选是通过表达蛋白上携带的6×His Tag经Ni-NTA柱纯化。
本发明所制备优化的猪瘟病毒E2蛋白鉴定的方法不受特别限制，如聚丙烯酰胺凝胶电泳、Western blotting等。
本发明的另一个目的在于提供含有如前所述优化的猪瘟病毒E2蛋白活性成分的血清学检测方法。
本发明获得的优化猪瘟病毒E2蛋白可用于制备猪瘟亚单位疫苗及制备猪瘟诊断试剂盒。
所述的优化的猪瘟病毒E2蛋白可用于制备检测伪狂犬病的血清学检测试剂盒。
本发明的优化的猪瘟病毒E2蛋白能够在哺乳动物细胞表达系统中高效表达，并可猪瘟毒株感染的猪的血清所特异性识别，猪瘟诊断具有应用价值。
附图说明
图1猪瘟病毒疫苗株E2蛋白基因GC分析图。
图2猪瘟病毒疫苗株E2蛋白密码子分析图。
图3猪瘟病毒疫苗株E2蛋白GC分析图。
图4猪瘟病毒疫苗株E2蛋白密码子分析图。
图5重组猪瘟病毒疫苗株E2蛋白电泳。
图6重组猪瘟病毒疫苗株E2蛋白与猪血清反应结果。
具体实施方式
实施例1猪瘟病毒疫苗株E2蛋白基因分析及编码的优化与改造
对猪瘟病毒疫苗株E2蛋白基因进行GC含量及分布进行分析，评估E2蛋白基因中密码子在哺乳动物细胞表达系统中的翻译效率，结果显示E2蛋白基因中GC在整个序列中平均分布在50％(如图1所示)，其中仅有50％的密码子可在哺乳动物细胞表达系统高效翻译，部分序列中CG含量低于40％(如图2所示)，说明猪瘟病毒疫苗株E2蛋白基因在哺乳动物细胞中表达效率不高；本发明根据哺乳动物细胞对密码子的选择性，将猪瘟病毒疫苗株E2蛋白基因上不适合的翻译的密码子进行逐一优化，经过优化后，GC在整个序列中平均含量优化到60％(如图3所示)，95％的密码子在哺乳动物细胞表达系统中翻译效率在85％以上(如图4所示)；本发明中，为提高猪瘟病毒疫苗株E2蛋白结构稳定性及外泌性，将N末端的添加IL-1信号肽，并将333位的甲硫氨酸残基突变为精氨酸残基，最后形成一个优化的猪瘟病毒E2蛋白基因SEQ1，氨基酸序列为SEQ2。
实施例2重组优化的猪瘟病毒E2蛋白表达质粒pcDNA3.4-CSFV-E2的构建
将人工合成的优化的猪瘟病毒E2蛋白基因利用In-fusion试剂盒连入pcDNA3.4载体中。转入大肠杆菌JM109，将菌液涂布于LB琼脂板(含氨苄青100μg/ml，氯霉素34μg/ml)，37℃过夜培养后，挑取克隆，抽提重组质粒进行酶切鉴定，结果显示成功构建了优化的猪瘟病毒E2蛋白的重组质粒pcDNA3.4-CSFV-E2，由上海英俊生物有限公司对该重组质粒进行测序鉴定，并采用Vector NTI 10软件，推定氨基酸序列，其结果于原始设计的优化的猪瘟病毒E2蛋白基因一致。
实施例3，重组优化猪瘟病毒E2蛋白的表达及纯化
重组质粒pcDNA3.4-CSFV-E2通过3000脂质体转入中华仓鼠卵巢细胞CHO-K1中，通过G418两周筛选后，进行单克隆培养，收集克隆培养上清，利用ForteBioOctet K2系统通过Anti-HIS(HIS2)Biosensors检测培养上清中E2蛋白表达产量，筛选出高效稳定表达优化猪瘟病毒E2蛋白的细胞株。通过无血清驯化后，进行扩大化培养，收集培养上清，通过选用His结合树脂(Novagen,Madison,Wis.)进行提纯。每升培养中可获纯化的优化猪瘟病毒E2蛋白50mg。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)检测其纯度。经12.5％的SDS-PAGE检测，在分子量大约50kD处出现目的蛋白表达条带(如图5所示)。
实施例4重组蛋白敏感性与特异性检测
96孔酶标板每孔加入含0.5μg重组的优化猪瘟病毒E2蛋白的Coating Buffer 100μl，4℃湿盒中过夜。PBS(含0.05％Tween20)洗板后每孔加入含3％脱脂奶的PBS 400μl，湿盒中室温封闭1h，每孔中分别加入100μl猪瘟疫苗免疫猪或猪瘟病毒野毒株感染猪的血清(1:50稀释)，湿盒中室温孵育1h。PBS(含0.05％Tween20)洗板后每孔中加入100μl辣根过氧化物酶标记的山羊抗猪IgG(1:1000稀释)100μl，湿盒中室温孵育1h。PBS(含0.05％Tween20)洗板后加入显色液200μl/孔，室温下避光反应30min，最后每孔加入2M H2SO4 50μl终止显色反应。酶标仪490nm测定吸光度；
ELISA检测结果显示，重组的优化后的猪瘟病毒E2蛋白不仅可与经过猪瘟疫苗免疫的猪血清中抗体特异性结合，也可以与猪瘟野毒株感染猪血清中抗体特异性结合，结果说明优化的猪瘟病毒E2蛋白可以特异性的被抗猪瘟病毒抗体识别；以优化的猪瘟病毒E2蛋白对132份临床标本进行检测，与商业化阻断抗体检测方法比较，其准确率为92％，特异性为92.5％，敏感性为93.83％。
实施例5重组蛋白免疫活性检测
2只6周健康雄性BALB/c小鼠，将150ug纯化后的猪瘟病毒E2蛋白与等体积的完全福氏佐剂混匀后，100ul/只腹腔注射；1周后以150ug纯化后的猪瘟病毒E2蛋白与等体积的不完全福氏佐剂混匀后，100ul/只腹腔注射；经过3次免疫后，小鼠剪尾采血，经检测免疫小鼠的血清效价高于1:240000，结果说明优化的猪瘟病毒E2蛋白可以诱导动物产生高滴度抗体。
。
序列表
<110> 复旦大学
<120> 一种人工密码子优化的猪瘟病毒E2蛋白制备与用途
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1251
<212> DNA
<213> pestivirus type 3
<220>
<221> CDS
<222> (4)..(1248)
<400> 1
atg tac agg atg cag ctg ctg agc tgt atc gcc ctg tct ctg gcc ctg 48
Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu
1 5 10 15
gtc aca aac tct gga cac cac cat cat cac cat cac cac cac cat agc 96
Val Thr Asn Ser Gly His His His His His His His His His His Ser
20 25 30
agc ggc cac atc gac gac gac gat aag ctg gaa aga ctg gcc tgc aaa 144
Ser Gly His Ile Asp Asp Asp Asp Lys Leu Glu Arg Leu Ala Cys Lys
35 40 45
gag gac tac aga tac gcc atc agc agc acc gac gag atc gga ttg ctt 192
Glu Asp Tyr Arg Tyr Ala Ile Ser Ser Thr Asp Glu Ile Gly Leu Leu
50 55 60
gga gct ggc ggc ctg acc acc acc tgg aaa gag tac aac cac gac ctg 240
Gly Ala Gly Gly Leu Thr Thr Thr Trp Lys Glu Tyr Asn His Asp Leu
65 70 75
cag ctg aac gac ggc aca gtg aag gcc tct tgt gtg gcc ggc agc ttc 288
Gln Leu Asn Asp Gly Thr Val Lys Ala Ser Cys Val Ala Gly Ser Phe
80 85 90 95
aaa gtg aca gcc ctg aac gtg gtg tct aga aga tac ctg gcc agc ctg 336
Lys Val Thr Ala Leu Asn Val Val Ser Arg Arg Tyr Leu Ala Ser Leu
100 105 110
cac aag aag gcc ctg cct aca agc gtg acc ttc gag ctg ctg ttc gac 384
His Lys Lys Ala Leu Pro Thr Ser Val Thr Phe Glu Leu Leu Phe Asp
115 120 125
ggc acc aat cct agc acc gag gaa atg ggc gac gac ttc aga tcc ggc 432
Gly Thr Asn Pro Ser Thr Glu Glu Met Gly Asp Asp Phe Arg Ser Gly
130 135 140
ctg tgt cct ttc gac aca agc cct gtg gtc aag ggc aag tac aac acc 480
Leu Cys Pro Phe Asp Thr Ser Pro Val Val Lys Gly Lys Tyr Asn Thr
145 150 155
aca ctg ctg aac ggc agc gcc ttc tac ctc gtg tgt cct atc gga tgg 528
Thr Leu Leu Asn Gly Ser Ala Phe Tyr Leu Val Cys Pro Ile Gly Trp
160 165 170 175
acc ggc gtg atc gag tgt aca gcc gtg tct cct acc aca ctg aga acc 576
Thr Gly Val Ile Glu Cys Thr Ala Val Ser Pro Thr Thr Leu Arg Thr
180 185 190
gag gtg gtc aag acc ttc aga agg gac aag ccc ttt cca cac aga atg 624
Glu Val Val Lys Thr Phe Arg Arg Asp Lys Pro Phe Pro His Arg Met
195 200 205
gac tgc gtg acc aca acc gtc gag aac gag gac ctg ttc tac tgc aag 672
Asp Cys Val Thr Thr Thr Val Glu Asn Glu Asp Leu Phe Tyr Cys Lys
210 215 220
ctc ggc ggc aac tgg acc tgt gtg aaa ggc gag cct gtg gtg tac aca 720
Leu Gly Gly Asn Trp Thr Cys Val Lys Gly Glu Pro Val Val Tyr Thr
225 230 235
ggc gga gtg gtt aag cag tgt cgt tgg tgc ggc ttc gac ttc gac gga 768
Gly Gly Val Val Lys Gln Cys Arg Trp Cys Gly Phe Asp Phe Asp Gly
240 245 250 255
cct gat gga ctg cct cac tac ccc atc ggc aag tgc atc ctg gct aac 816
Pro Asp Gly Leu Pro His Tyr Pro Ile Gly Lys Cys Ile Leu Ala Asn
260 265 270
gag aca ggc tac aga atc gtg gac tcc acc gac tgc aac aga gat ggc 864
Glu Thr Gly Tyr Arg Ile Val Asp Ser Thr Asp Cys Asn Arg Asp Gly
275 280 285
gtg gtc atc agc aca gag ggc agc cac gaa tgc ctg atc ggc aac aca 912
Val Val Ile Ser Thr Glu Gly Ser His Glu Cys Leu Ile Gly Asn Thr
290 295 300
acc gtg aag gtg cac gcc agc gac gag aga ctt ggc cct atg cct tgc 960
Thr Val Lys Val His Ala Ser Asp Glu Arg Leu Gly Pro Met Pro Cys
305 310 315
aga ccc aaa gaa atc gtg tct agc gct ggc ccc gtg cgc aag acc agc 1008
Arg Pro Lys Glu Ile Val Ser Ser Ala Gly Pro Val Arg Lys Thr Ser
320 325 330 335
tgt acc ttc aac tac aca aag acc ctg aag aac cgg tac tac gag ccc 1056
Cys Thr Phe Asn Tyr Thr Lys Thr Leu Lys Asn Arg Tyr Tyr Glu Pro
340 345 350
agg gac agc tac ttc cag cag tac atg ctg aag ggc gag tac cag tat 1104
Arg Asp Ser Tyr Phe Gln Gln Tyr Met Leu Lys Gly Glu Tyr Gln Tyr
355 360 365
tgg ttc gac ctg gac gcc aca gac aga cac agc gac tac ttc gcc gag 1152
Trp Phe Asp Leu Asp Ala Thr Asp Arg His Ser Asp Tyr Phe Ala Glu
370 375 380
ttc gtg gtg ctg gtg gtt gtg gca ctt ctc ggc gga aga tac gtg ctg 1200
Phe Val Val Leu Val Val Val Ala Leu Leu Gly Gly Arg Tyr Val Leu
385 390 395
tgg ctg atc gtg acc tat gtg gtg ctg acc gag cag ctg gct gct gga tga 1251
Trp Leu Ile Val Thr Tyr Val Val Leu Thr Glu Gln Leu Ala Ala Gly
400 405 410 415
<210> 2
<211> 415
<212> PRT
<213> pestivirus type 3
<400> 2
Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu Val
1 5 10 15
Thr Asn Ser Gly His His His His His His His His His His Ser Ser
20 25 30
Gly His Ile Asp Asp Asp Asp Lys Leu Glu Arg Leu Ala Cys Lys Glu
35 40 45
Asp Tyr Arg Tyr Ala Ile Ser Ser Thr Asp Glu Ile Gly Leu Leu Gly
50 55 60
Ala Gly Gly Leu Thr Thr Thr Trp Lys Glu Tyr Asn His Asp Leu Gln
65 70 75 80
Leu Asn Asp Gly Thr Val Lys Ala Ser Cys Val Ala Gly Ser Phe Lys
85 90 95
Val Thr Ala Leu Asn Val Val Ser Arg Arg Tyr Leu Ala Ser Leu His
100 105 110
Lys Lys Ala Leu Pro Thr Ser Val Thr Phe Glu Leu Leu Phe Asp Gly
115 120 125
Thr Asn Pro Ser Thr Glu Glu Met Gly Asp Asp Phe Arg Ser Gly Leu
130 135 140
Cys Pro Phe Asp Thr Ser Pro Val Val Lys Gly Lys Tyr Asn Thr Thr
145 150 155 160
Leu Leu Asn Gly Ser Ala Phe Tyr Leu Val Cys Pro Ile Gly Trp Thr
165 170 175
Gly Val Ile Glu Cys Thr Ala Val Ser Pro Thr Thr Leu Arg Thr Glu
180 185 190
Val Val Lys Thr Phe Arg Arg Asp Lys Pro Phe Pro His Arg Met Asp
195 200 205
Cys Val Thr Thr Thr Val Glu Asn Glu Asp Leu Phe Tyr Cys Lys Leu
210 215 220
Gly Gly Asn Trp Thr Cys Val Lys Gly Glu Pro Val Val Tyr Thr Gly
225 230 235 240
Gly Val Val Lys Gln Cys Arg Trp Cys Gly Phe Asp Phe Asp Gly Pro
245 250 255
Asp Gly Leu Pro His Tyr Pro Ile Gly Lys Cys Ile Leu Ala Asn Glu
260 265 270
Thr Gly Tyr Arg Ile Val Asp Ser Thr Asp Cys Asn Arg Asp Gly Val
275 280 285
Val Ile Ser Thr Glu Gly Ser His Glu Cys Leu Ile Gly Asn Thr Thr
290 295 300
Val Lys Val His Ala Ser Asp Glu Arg Leu Gly Pro Met Pro Cys Arg
305 310 315 320
Pro Lys Glu Ile Val Ser Ser Ala Gly Pro Val Arg Lys Thr Ser Cys
325 330 335
Thr Phe Asn Tyr Thr Lys Thr Leu Lys Asn Arg Tyr Tyr Glu Pro Arg
340 345 350
Asp Ser Tyr Phe Gln Gln Tyr Met Leu Lys Gly Glu Tyr Gln Tyr Trp
355 360 365
Phe Asp Leu Asp Ala Thr Asp Arg His Ser Asp Tyr Phe Ala Glu Phe
370 375 380
Val Val Leu Val Val Val Ala Leu Leu Gly Gly Arg Tyr Val Leu Trp
385 390 395 400
Leu Ile Val Thr Tyr Val Val Leu Thr Glu Gln Leu Ala Ala Gly
405 410 415
Claims (4)
1. 一种优化猪瘟病毒E2 蛋白的基因，其特征在于，其核苷酸序列如SEQ ID NO.1所示。
2. 一种表达载体，其特征在于，含有权利要求1所述的优化的猪瘟病毒E2 蛋白的基因。
3.根据权利要求2所述的表达载体，其特征在于，所述的表达载体是质粒pcDNA3.4。
4.一种含有权利要求3 所述的表达载体的宿主细胞，其特征在于，所述的宿主细胞是哺乳动物细胞。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911122079.4A CN112813080B (zh) | 2019-11-15 | 2019-11-15 | 一种人工密码子优化的猪瘟病毒e2蛋白及其制备方法与用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911122079.4A CN112813080B (zh) | 2019-11-15 | 2019-11-15 | 一种人工密码子优化的猪瘟病毒e2蛋白及其制备方法与用途 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112813080A CN112813080A (zh) | 2021-05-18 |
| CN112813080B true CN112813080B (zh) | 2023-12-22 |
Family
ID=75851902
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911122079.4A Active CN112813080B (zh) | 2019-11-15 | 2019-11-15 | 一种人工密码子优化的猪瘟病毒e2蛋白及其制备方法与用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112813080B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115586338A (zh) * | 2022-09-20 | 2023-01-10 | 杭州赛基生物科技有限公司 | 用于检测可溶性细胞因子受体的试剂盒及其制备方法 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103751774A (zh) * | 2013-07-17 | 2014-04-30 | 哈尔滨维科生物技术开发公司 | 稳定表达猪瘟病毒e2蛋白的重组细胞系及其在制备猪瘟亚单位疫苗和诊断试剂中的应用 |
| CN107674883A (zh) * | 2016-08-01 | 2018-02-09 | 浙江海隆生物科技有限公司 | 重组猪瘟e2蛋白及其亚单位疫苗的制备方法和应用 |
| CN108395996A (zh) * | 2018-01-31 | 2018-08-14 | 复旦大学 | 一种猪瘟病毒亚单位疫苗及其制备方法和用途 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9359411B2 (en) * | 2008-07-31 | 2016-06-07 | Maw Hsing Biotech Co., Ltd. | Yeast expressed classical swine fever virus glycoprotein E2 and use thereof |
-
2019
- 2019-11-15 CN CN201911122079.4A patent/CN112813080B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103751774A (zh) * | 2013-07-17 | 2014-04-30 | 哈尔滨维科生物技术开发公司 | 稳定表达猪瘟病毒e2蛋白的重组细胞系及其在制备猪瘟亚单位疫苗和诊断试剂中的应用 |
| CN107674883A (zh) * | 2016-08-01 | 2018-02-09 | 浙江海隆生物科技有限公司 | 重组猪瘟e2蛋白及其亚单位疫苗的制备方法和应用 |
| CN108395996A (zh) * | 2018-01-31 | 2018-08-14 | 复旦大学 | 一种猪瘟病毒亚单位疫苗及其制备方法和用途 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112813080A (zh) | 2021-05-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113861278B (zh) | 一种产生广谱交叉中和活性的重组新型冠状病毒rbd三聚体蛋白疫苗、其制备方法和应用 | |
| JP5829830B2 (ja) | Ｅ型肝炎ウイルスのポリペプチド断片、それを含むワクチン組成物及び診断キット、並びにその使用 | |
| CN112111005B (zh) | 一种能与阳性血清竞争结合非洲猪瘟病毒b646l抗原的单克隆抗体及其应用 | |
| CN115043948B (zh) | 一种猴痘病毒特异性融合蛋白抗原及其制备方法和应用 | |
| CN114315989A (zh) | 一种重组新型冠状病毒蛋白疫苗、其制备方法和应用 | |
| CN111116720A (zh) | 一种猪瘟病毒重组e2蛋白及其应用 | |
| CN112920278A (zh) | 一种新型冠状病毒特异性融合蛋白抗原及其制备方法和应用 | |
| CN114478718A (zh) | 一种重组新型冠状病毒蛋白疫苗、其制备方法和应用 | |
| CN112813080B (zh) | 一种人工密码子优化的猪瘟病毒e2蛋白及其制备方法与用途 | |
| CN116655750A (zh) | 一种prrsv m蛋白sla-1*04:01:01限制性t细胞表位多肽及其应用 | |
| CN114933639B (zh) | 非洲猪瘟病毒p72N抗原表位蛋白及其制备方法和用途 | |
| CN114478717B (zh) | 一种重组新型冠状病毒蛋白疫苗、其制备方法和应用 | |
| CN114409742B (zh) | 非洲猪瘟病毒p49蛋白抗原表位及其应用 | |
| CN112724205B (zh) | C种戊型肝炎病毒239蛋白制备病毒样颗粒的方法和应用 | |
| KR102484312B1 (ko) | 고효능 사스 코로나바이러스 2 항원 및 이를 포함하는 백신 조성물 | |
| CN114315984A (zh) | 一种用于制备prrsv基因ii型表位缺失疫苗毒株的n蛋白表位突变标记及其应用 | |
| CN110563816B (zh) | 一种人工密码子优化的伪狂犬病毒类gE蛋白及其用途 | |
| CN107619435B (zh) | 一种猪瘟病毒e2蛋白的抗原表位、抗体的制备及应用 | |
| US10117913B2 (en) | Antigens of Pneumocystis murina and uses thereof | |
| CN113372454B (zh) | 尼帕病毒受体结合糖蛋白及其应用 | |
| Wu et al. | Characterization of the monoclonal antibody against classical swine fever virus glycoprotein E rns and its application to an indirect sandwich ELISA | |
| JPH10505236A (ja) | ネコ科の感染性腹膜炎用ワクチン | |
| 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 | |
| CN113493494B (zh) | Eb病毒balf3蛋白的抗原表位 | |
| CN103740736B (zh) | 化学合成的HSV2病毒gE糖蛋白胞外区基因片段及其表达、应用 |
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 |