CN111607001B - 一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白及其制备方法和应用 - Google Patents
一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白及其制备方法和应用 Download PDFInfo
- Publication number
- CN111607001B CN111607001B CN201910142648.5A CN201910142648A CN111607001B CN 111607001 B CN111607001 B CN 111607001B CN 201910142648 A CN201910142648 A CN 201910142648A CN 111607001 B CN111607001 B CN 111607001B
- Authority
- CN
- China
- Prior art keywords
- protein
- opti
- swine fever
- african swine
- fever virus
- 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
- 241000701386 African swine fever virus Species 0.000 title claims abstract description 35
- 108020001507 fusion proteins Proteins 0.000 title claims abstract description 23
- 102000037865 fusion proteins Human genes 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 78
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 71
- 239000002773 nucleotide Substances 0.000 claims abstract description 23
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 23
- 241000588724 Escherichia coli Species 0.000 claims abstract description 16
- 102000005431 Molecular Chaperones Human genes 0.000 claims abstract description 9
- 108010006519 Molecular Chaperones Proteins 0.000 claims abstract description 9
- 101710121996 Hexon protein p72 Proteins 0.000 claims description 25
- 101710125418 Major capsid protein Proteins 0.000 claims description 25
- 230000014509 gene expression Effects 0.000 claims description 23
- 238000000034 method Methods 0.000 claims description 18
- 208000007407 African swine fever Diseases 0.000 claims description 17
- 239000013612 plasmid Substances 0.000 claims description 17
- 229960005486 vaccine Drugs 0.000 claims description 17
- 239000013604 expression vector Substances 0.000 claims description 15
- 238000012258 culturing Methods 0.000 claims description 13
- 230000009465 prokaryotic expression Effects 0.000 claims description 10
- 210000004027 cell Anatomy 0.000 claims description 7
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 6
- 108050001186 Chaperonin Cpn60 Proteins 0.000 claims description 5
- 102000052603 Chaperonins Human genes 0.000 claims description 5
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 5
- 101710135898 Myc proto-oncogene protein Proteins 0.000 claims description 4
- 102100038895 Myc proto-oncogene protein Human genes 0.000 claims description 4
- 101710150448 Transcriptional regulator Myc Proteins 0.000 claims description 4
- 238000003745 diagnosis Methods 0.000 claims description 3
- 238000010367 cloning Methods 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims description 2
- 238000003259 recombinant expression Methods 0.000 claims description 2
- 238000012216 screening Methods 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 210000004899 c-terminal region Anatomy 0.000 claims 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 29
- 101800000958 Reverse transcriptase/ribonuclease H Proteins 0.000 description 26
- 238000000855 fermentation Methods 0.000 description 21
- 230000004151 fermentation Effects 0.000 description 21
- 238000006243 chemical reaction Methods 0.000 description 12
- 239000000243 solution Substances 0.000 description 12
- 239000013598 vector Substances 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 11
- 230000001580 bacterial effect Effects 0.000 description 11
- 235000011187 glycerol Nutrition 0.000 description 10
- 108020004414 DNA Proteins 0.000 description 9
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 9
- 230000006698 induction Effects 0.000 description 9
- 239000007788 liquid Substances 0.000 description 8
- 238000005457 optimization Methods 0.000 description 8
- 210000002966 serum Anatomy 0.000 description 8
- 150000001413 amino acids Chemical group 0.000 description 7
- 239000012071 phase Substances 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 6
- 241000700605 Viruses Species 0.000 description 6
- 239000011248 coating agent Substances 0.000 description 6
- 238000000576 coating method Methods 0.000 description 6
- 239000000306 component Substances 0.000 description 6
- 239000012533 medium component Substances 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 108020004705 Codon Proteins 0.000 description 5
- 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 5
- 240000000220 Panda oleosa Species 0.000 description 5
- 235000016496 Panda oleosa Nutrition 0.000 description 5
- 239000000427 antigen Substances 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 239000012149 elution buffer Substances 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 208000015181 infectious disease Diseases 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 238000007789 sealing Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- IDOQDZANRZQBTP-UHFFFAOYSA-N 2-[2-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol Chemical compound CC(C)(C)CC(C)(C)C1=CC=CC=C1OCCO IDOQDZANRZQBTP-UHFFFAOYSA-N 0.000 description 4
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 4
- 241000282887 Suidae Species 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 238000001976 enzyme digestion Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000012526 feed medium Substances 0.000 description 4
- 230000003053 immunization Effects 0.000 description 4
- 238000002649 immunization Methods 0.000 description 4
- 238000010255 intramuscular injection Methods 0.000 description 4
- 239000007927 intramuscular injection Substances 0.000 description 4
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 4
- 238000011068 loading method Methods 0.000 description 4
- 239000006166 lysate Substances 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000012474 protein marker Substances 0.000 description 4
- 229940031626 subunit vaccine Drugs 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 230000009466 transformation Effects 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 241000880493 Leptailurus serval Species 0.000 description 3
- 241001052560 Thallis Species 0.000 description 3
- 229920004929 Triton X-114 Polymers 0.000 description 3
- 108010047857 aspartylglycine Proteins 0.000 description 3
- 238000010276 construction Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 108010009298 lysylglutamic acid Proteins 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 238000001742 protein purification Methods 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- SEKBHZJLARBNPB-GHCJXIJMSA-N Asn-Ile-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O SEKBHZJLARBNPB-GHCJXIJMSA-N 0.000 description 2
- QNMKWNONJGKJJC-NHCYSSNCSA-N Asp-Leu-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O QNMKWNONJGKJJC-NHCYSSNCSA-N 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- LZEUDRYSAZAJIO-AUTRQRHGSA-N Glu-Val-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LZEUDRYSAZAJIO-AUTRQRHGSA-N 0.000 description 2
- DHDOADIPGZTAHT-YUMQZZPRSA-N Gly-Glu-Arg Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DHDOADIPGZTAHT-YUMQZZPRSA-N 0.000 description 2
- LLZLRXBTOOFODM-QSFUFRPTSA-N Ile-Asp-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C(C)C)C(=O)O)N LLZLRXBTOOFODM-QSFUFRPTSA-N 0.000 description 2
- CSQNHSGHAPRGPQ-YTFOTSKYSA-N Ile-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)O)N CSQNHSGHAPRGPQ-YTFOTSKYSA-N 0.000 description 2
- KCTIFOCXAIUQQK-QXEWZRGKSA-N Ile-Pro-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O KCTIFOCXAIUQQK-QXEWZRGKSA-N 0.000 description 2
- 108010065920 Insulin Lispro Proteins 0.000 description 2
- AKVBOOKXVAMKSS-GUBZILKMSA-N Leu-Ser-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O AKVBOOKXVAMKSS-GUBZILKMSA-N 0.000 description 2
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 2
- KIAWKQJTSGRCSA-AVGNSLFASA-N Phe-Asn-Glu Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N KIAWKQJTSGRCSA-AVGNSLFASA-N 0.000 description 2
- SSWJYJHXQOYTSP-SRVKXCTJSA-N Pro-His-Gln Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(N)=O)C(O)=O SSWJYJHXQOYTSP-SRVKXCTJSA-N 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 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 2
- 241000282898 Sus scrofa Species 0.000 description 2
- VIBXMCZWVUOZLA-OLHMAJIHSA-N Thr-Asn-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O VIBXMCZWVUOZLA-OLHMAJIHSA-N 0.000 description 2
- LMMDEZPNUTZJAY-GCJQMDKQSA-N Thr-Asp-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O LMMDEZPNUTZJAY-GCJQMDKQSA-N 0.000 description 2
- JMBRNXUOLJFURW-BEAPCOKYSA-N Thr-Phe-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N)O JMBRNXUOLJFURW-BEAPCOKYSA-N 0.000 description 2
- YVNQAIFQFWTPLQ-UHFFFAOYSA-O [4-[[4-(4-ethoxyanilino)phenyl]-[4-[ethyl-[(3-sulfophenyl)methyl]amino]-2-methylphenyl]methylidene]-3-methylcyclohexa-2,5-dien-1-ylidene]-ethyl-[(3-sulfophenyl)methyl]azanium Chemical compound C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C(=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S(O)(=O)=O)C)C=2C(=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S(O)(=O)=O)C)C=C1 YVNQAIFQFWTPLQ-UHFFFAOYSA-O 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 108010041407 alanylaspartic acid Proteins 0.000 description 2
- 108010044940 alanylglutamine Proteins 0.000 description 2
- 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 2
- 108010077245 asparaginyl-proline Proteins 0.000 description 2
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000029142 excretion Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 2
- 108010015792 glycyllysine Proteins 0.000 description 2
- 210000003000 inclusion body Anatomy 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 108010057821 leucylproline Proteins 0.000 description 2
- 108010003700 lysyl aspartic acid Proteins 0.000 description 2
- 229910052759 nickel Inorganic materials 0.000 description 2
- 230000000704 physical effect Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 108010031719 prolyl-serine Proteins 0.000 description 2
- 108010029020 prolylglycine Proteins 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 108010051110 tyrosyl-lysine Proteins 0.000 description 2
- 108010073969 valyllysine Proteins 0.000 description 2
- 239000011534 wash buffer Substances 0.000 description 2
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 1
- 101900228950 African swine fever virus Phosphoprotein p30 Proteins 0.000 description 1
- SVBXIUDNTRTKHE-CIUDSAMLSA-N Ala-Arg-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O SVBXIUDNTRTKHE-CIUDSAMLSA-N 0.000 description 1
- GFBLJMHGHAXGNY-ZLUOBGJFSA-N Ala-Asn-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O GFBLJMHGHAXGNY-ZLUOBGJFSA-N 0.000 description 1
- BUDNAJYVCUHLSV-ZLUOBGJFSA-N Ala-Asp-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O BUDNAJYVCUHLSV-ZLUOBGJFSA-N 0.000 description 1
- CXQODNIBUNQWAS-CIUDSAMLSA-N Ala-Gln-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N CXQODNIBUNQWAS-CIUDSAMLSA-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
- GGNHBHYDMUDXQB-KBIXCLLPSA-N Ala-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)N GGNHBHYDMUDXQB-KBIXCLLPSA-N 0.000 description 1
- OMMDTNGURYRDAC-NRPADANISA-N Ala-Glu-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O OMMDTNGURYRDAC-NRPADANISA-N 0.000 description 1
- BEMGNWZECGIJOI-WDSKDSINSA-N Ala-Gly-Glu Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O BEMGNWZECGIJOI-WDSKDSINSA-N 0.000 description 1
- PCIFXPRIFWKWLK-YUMQZZPRSA-N Ala-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N PCIFXPRIFWKWLK-YUMQZZPRSA-N 0.000 description 1
- FAJIYNONGXEXAI-CQDKDKBSSA-N Ala-His-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CNC=N1 FAJIYNONGXEXAI-CQDKDKBSSA-N 0.000 description 1
- CKLDHDOIYBVUNP-KBIXCLLPSA-N Ala-Ile-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O CKLDHDOIYBVUNP-KBIXCLLPSA-N 0.000 description 1
- OKIKVSXTXVVFDV-MMWGEVLESA-N Ala-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C)N OKIKVSXTXVVFDV-MMWGEVLESA-N 0.000 description 1
- UWIQWPWWZUHBAO-ZLIFDBKOSA-N Ala-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@H](C)N)CC(C)C)C(O)=O)=CNC2=C1 UWIQWPWWZUHBAO-ZLIFDBKOSA-N 0.000 description 1
- VCSABYLVNWQYQE-UHFFFAOYSA-N Ala-Lys-Lys Natural products NCCCCC(NC(=O)C(N)C)C(=O)NC(CCCCN)C(O)=O VCSABYLVNWQYQE-UHFFFAOYSA-N 0.000 description 1
- MDNAVFBZPROEHO-DCAQKATOSA-N Ala-Lys-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O MDNAVFBZPROEHO-DCAQKATOSA-N 0.000 description 1
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 1
- OMDNCNKNEGFOMM-BQBZGAKWSA-N Ala-Met-Gly Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O OMDNCNKNEGFOMM-BQBZGAKWSA-N 0.000 description 1
- WQLDNOCHHRISMS-NAKRPEOUSA-N Ala-Pro-Ile Chemical compound [H]N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WQLDNOCHHRISMS-NAKRPEOUSA-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
- LTTLSZVJTDSACD-OWLDWWDNSA-N Ala-Thr-Trp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O LTTLSZVJTDSACD-OWLDWWDNSA-N 0.000 description 1
- VWVPYNGMOCSSGK-GUBZILKMSA-N Arg-Arg-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O VWVPYNGMOCSSGK-GUBZILKMSA-N 0.000 description 1
- NONSEUUPKITYQT-BQBZGAKWSA-N Arg-Asn-Gly Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)O)N)CN=C(N)N NONSEUUPKITYQT-BQBZGAKWSA-N 0.000 description 1
- BVBKBQRPOJFCQM-DCAQKATOSA-N Arg-Asn-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O BVBKBQRPOJFCQM-DCAQKATOSA-N 0.000 description 1
- ITVINTQUZMQWJR-QXEWZRGKSA-N Arg-Asn-Val Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O ITVINTQUZMQWJR-QXEWZRGKSA-N 0.000 description 1
- FEZJJKXNPSEYEV-CIUDSAMLSA-N Arg-Gln-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O FEZJJKXNPSEYEV-CIUDSAMLSA-N 0.000 description 1
- PTVGLOCPAVYPFG-CIUDSAMLSA-N Arg-Gln-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O PTVGLOCPAVYPFG-CIUDSAMLSA-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
- YKBHOXLMMPZPHQ-GMOBBJLQSA-N Arg-Ile-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O YKBHOXLMMPZPHQ-GMOBBJLQSA-N 0.000 description 1
- SSZGOKWBHLOCHK-DCAQKATOSA-N Arg-Lys-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCN=C(N)N SSZGOKWBHLOCHK-DCAQKATOSA-N 0.000 description 1
- XUGATJVGQUGQKY-ULQDDVLXSA-N Arg-Lys-Phe Chemical compound NC(=N)NCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 XUGATJVGQUGQKY-ULQDDVLXSA-N 0.000 description 1
- GSUFZRURORXYTM-STQMWFEESA-N Arg-Phe-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=CC=C1 GSUFZRURORXYTM-STQMWFEESA-N 0.000 description 1
- KZXPVYVSHUJCEO-ULQDDVLXSA-N Arg-Phe-Lys Chemical compound NC(=N)NCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCCN)C(O)=O)CC1=CC=CC=C1 KZXPVYVSHUJCEO-ULQDDVLXSA-N 0.000 description 1
- XFXZKCRBBOVJKS-BVSLBCMMSA-N Arg-Phe-Trp Chemical compound C([C@H](NC(=O)[C@H](CCCN=C(N)N)N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=CC=C1 XFXZKCRBBOVJKS-BVSLBCMMSA-N 0.000 description 1
- STHNZYKCJHWULY-AVGNSLFASA-N Arg-Pro-His Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CCCN=C(N)N)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O STHNZYKCJHWULY-AVGNSLFASA-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
- AKEBUSZTMQLNIX-UWJYBYFXSA-N Asn-Ala-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N AKEBUSZTMQLNIX-UWJYBYFXSA-N 0.000 description 1
- JEPNYDRDYNSFIU-QXEWZRGKSA-N Asn-Arg-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(N)=O)C(O)=O JEPNYDRDYNSFIU-QXEWZRGKSA-N 0.000 description 1
- OLGCWMNDJTWQAG-GUBZILKMSA-N Asn-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(N)=O OLGCWMNDJTWQAG-GUBZILKMSA-N 0.000 description 1
- COUZKSSMBFADSB-AVGNSLFASA-N Asn-Glu-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC(=O)N)N COUZKSSMBFADSB-AVGNSLFASA-N 0.000 description 1
- DXVMJJNAOVECBA-WHFBIAKZSA-N Asn-Gly-Asn Chemical compound NC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O DXVMJJNAOVECBA-WHFBIAKZSA-N 0.000 description 1
- GJFYPBDMUGGLFR-NKWVEPMBSA-N Asn-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CC(=O)N)N)C(=O)O GJFYPBDMUGGLFR-NKWVEPMBSA-N 0.000 description 1
- ZTRJUKDEALVRMW-SRVKXCTJSA-N Asn-His-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CC(=O)N)N ZTRJUKDEALVRMW-SRVKXCTJSA-N 0.000 description 1
- PHJPKNUWWHRAOC-PEFMBERDSA-N Asn-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N PHJPKNUWWHRAOC-PEFMBERDSA-N 0.000 description 1
- KMCRKVOLRCOMBG-DJFWLOJKSA-N Asn-Ile-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC(=O)N)N KMCRKVOLRCOMBG-DJFWLOJKSA-N 0.000 description 1
- BZWRLDPIWKOVKB-ZPFDUUQYSA-N Asn-Leu-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BZWRLDPIWKOVKB-ZPFDUUQYSA-N 0.000 description 1
- YVXRYLVELQYAEQ-SRVKXCTJSA-N Asn-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N YVXRYLVELQYAEQ-SRVKXCTJSA-N 0.000 description 1
- ORJQQZIXTOYGGH-SRVKXCTJSA-N Asn-Lys-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O ORJQQZIXTOYGGH-SRVKXCTJSA-N 0.000 description 1
- NLDNNZKUSLAYFW-NHCYSSNCSA-N Asn-Lys-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O NLDNNZKUSLAYFW-NHCYSSNCSA-N 0.000 description 1
- PLTGTJAZQRGMPP-FXQIFTODSA-N Asn-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC(N)=O PLTGTJAZQRGMPP-FXQIFTODSA-N 0.000 description 1
- OOXUBGLNDRGOKT-FXQIFTODSA-N Asn-Ser-Arg Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O OOXUBGLNDRGOKT-FXQIFTODSA-N 0.000 description 1
- DOURAOODTFJRIC-CIUDSAMLSA-N Asn-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)N DOURAOODTFJRIC-CIUDSAMLSA-N 0.000 description 1
- CBWCQCANJSGUOH-ZKWXMUAHSA-N Asn-Val-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O CBWCQCANJSGUOH-ZKWXMUAHSA-N 0.000 description 1
- XLDMSQYOYXINSZ-QXEWZRGKSA-N Asn-Val-Arg Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N XLDMSQYOYXINSZ-QXEWZRGKSA-N 0.000 description 1
- MJIJBEYEHBKTIM-BYULHYEWSA-N Asn-Val-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N MJIJBEYEHBKTIM-BYULHYEWSA-N 0.000 description 1
- ZLGKHJHFYSRUBH-FXQIFTODSA-N Asp-Arg-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O ZLGKHJHFYSRUBH-FXQIFTODSA-N 0.000 description 1
- XYBJLTKSGFBLCS-QXEWZRGKSA-N Asp-Arg-Val Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)CC(O)=O XYBJLTKSGFBLCS-QXEWZRGKSA-N 0.000 description 1
- ATYWBXGNXZYZGI-ACZMJKKPSA-N Asp-Asn-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O ATYWBXGNXZYZGI-ACZMJKKPSA-N 0.000 description 1
- WLKVEEODTPQPLI-ACZMJKKPSA-N Asp-Gln-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O WLKVEEODTPQPLI-ACZMJKKPSA-N 0.000 description 1
- XJQRWGXKUSDEFI-ACZMJKKPSA-N Asp-Glu-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O XJQRWGXKUSDEFI-ACZMJKKPSA-N 0.000 description 1
- KHBLRHKVXICFMY-GUBZILKMSA-N Asp-Glu-Lys Chemical compound N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O KHBLRHKVXICFMY-GUBZILKMSA-N 0.000 description 1
- PSLSTUMPZILTAH-BYULHYEWSA-N Asp-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(O)=O PSLSTUMPZILTAH-BYULHYEWSA-N 0.000 description 1
- GBSUGIXJAAKZOW-GMOBBJLQSA-N Asp-Ile-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O GBSUGIXJAAKZOW-GMOBBJLQSA-N 0.000 description 1
- UJGRZQYSNYTCAX-SRVKXCTJSA-N Asp-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UJGRZQYSNYTCAX-SRVKXCTJSA-N 0.000 description 1
- KFAFUJMGHVVYRC-DCAQKATOSA-N Asp-Leu-Met Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(O)=O KFAFUJMGHVVYRC-DCAQKATOSA-N 0.000 description 1
- HJCGDIGVVWETRO-ZPFDUUQYSA-N Asp-Lys-Ile Chemical compound CC[C@H](C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC(O)=O)C(O)=O HJCGDIGVVWETRO-ZPFDUUQYSA-N 0.000 description 1
- FQHBAQLBIXLWAG-DCAQKATOSA-N Asp-Lys-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(=O)O)N FQHBAQLBIXLWAG-DCAQKATOSA-N 0.000 description 1
- GPPIDDWYKJPRES-YDHLFZDLSA-N Asp-Phe-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O GPPIDDWYKJPRES-YDHLFZDLSA-N 0.000 description 1
- UAXIKORUDGGIGA-DCAQKATOSA-N Asp-Pro-Lys Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC(=O)O)N)C(=O)N[C@@H](CCCCN)C(=O)O UAXIKORUDGGIGA-DCAQKATOSA-N 0.000 description 1
- WAEDSQFVZJUHLI-BYULHYEWSA-N Asp-Val-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O WAEDSQFVZJUHLI-BYULHYEWSA-N 0.000 description 1
- GIKOVDMXBAFXDF-NHCYSSNCSA-N Asp-Val-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O GIKOVDMXBAFXDF-NHCYSSNCSA-N 0.000 description 1
- RKXVTTIQNKPCHU-KKHAAJSZSA-N Asp-Val-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC(O)=O RKXVTTIQNKPCHU-KKHAAJSZSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 108700010070 Codon Usage Proteins 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- AOZBJZBKFHOYHL-AVGNSLFASA-N Cys-Glu-Tyr Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O AOZBJZBKFHOYHL-AVGNSLFASA-N 0.000 description 1
- WAJDEKCJRKGRPG-CIUDSAMLSA-N Cys-His-Ser Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CS)N WAJDEKCJRKGRPG-CIUDSAMLSA-N 0.000 description 1
- 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 1
- YNJBLTDKTMKEET-ZLUOBGJFSA-N Cys-Ser-Ser Chemical compound SC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O YNJBLTDKTMKEET-ZLUOBGJFSA-N 0.000 description 1
- KVYVOGYEMPEXBT-GUBZILKMSA-N Gln-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O KVYVOGYEMPEXBT-GUBZILKMSA-N 0.000 description 1
- SHERTACNJPYHAR-ACZMJKKPSA-N Gln-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O SHERTACNJPYHAR-ACZMJKKPSA-N 0.000 description 1
- MQANCSUBSBJNLU-KKUMJFAQSA-N Gln-Arg-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O MQANCSUBSBJNLU-KKUMJFAQSA-N 0.000 description 1
- IKFZXRLDMYWNBU-YUMQZZPRSA-N Gln-Gly-Arg Chemical compound NC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N IKFZXRLDMYWNBU-YUMQZZPRSA-N 0.000 description 1
- JXFLPKSDLDEOQK-JHEQGTHGSA-N Gln-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCC(N)=O JXFLPKSDLDEOQK-JHEQGTHGSA-N 0.000 description 1
- WLRYGVYQFXRJDA-DCAQKATOSA-N Gln-Pro-Pro Chemical compound NC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 WLRYGVYQFXRJDA-DCAQKATOSA-N 0.000 description 1
- VNTGPISAOMAXRK-CIUDSAMLSA-N Gln-Pro-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O VNTGPISAOMAXRK-CIUDSAMLSA-N 0.000 description 1
- VLOLPWWCNKWRNB-LOKLDPHHSA-N Gln-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O VLOLPWWCNKWRNB-LOKLDPHHSA-N 0.000 description 1
- FITIQFSXXBKFFM-NRPADANISA-N Gln-Val-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O FITIQFSXXBKFFM-NRPADANISA-N 0.000 description 1
- LKDIBBOKUAASNP-FXQIFTODSA-N Glu-Ala-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LKDIBBOKUAASNP-FXQIFTODSA-N 0.000 description 1
- CGYDXNKRIMJMLV-GUBZILKMSA-N Glu-Arg-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O CGYDXNKRIMJMLV-GUBZILKMSA-N 0.000 description 1
- ZOXBSICWUDAOHX-GUBZILKMSA-N Glu-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCC(O)=O ZOXBSICWUDAOHX-GUBZILKMSA-N 0.000 description 1
- PAQUJCSYVIBPLC-AVGNSLFASA-N Glu-Asp-Phe Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 PAQUJCSYVIBPLC-AVGNSLFASA-N 0.000 description 1
- YLJHCWNDBKKOEB-IHRRRGAJSA-N Glu-Glu-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O YLJHCWNDBKKOEB-IHRRRGAJSA-N 0.000 description 1
- BUAKRRKDHSSIKK-IHRRRGAJSA-N Glu-Glu-Tyr Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 BUAKRRKDHSSIKK-IHRRRGAJSA-N 0.000 description 1
- PXXGVUVQWQGGIG-YUMQZZPRSA-N Glu-Gly-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N PXXGVUVQWQGGIG-YUMQZZPRSA-N 0.000 description 1
- OAGVHWYIBZMWLA-YFKPBYRVSA-N Glu-Gly-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)NCC(O)=O OAGVHWYIBZMWLA-YFKPBYRVSA-N 0.000 description 1
- LRPXYSGPOBVBEH-IUCAKERBSA-N Glu-Gly-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O LRPXYSGPOBVBEH-IUCAKERBSA-N 0.000 description 1
- HPJLZFTUUJKWAJ-JHEQGTHGSA-N Glu-Gly-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HPJLZFTUUJKWAJ-JHEQGTHGSA-N 0.000 description 1
- ZHNHJYYFCGUZNQ-KBIXCLLPSA-N Glu-Ile-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCC(O)=O ZHNHJYYFCGUZNQ-KBIXCLLPSA-N 0.000 description 1
- IVGJYOOGJLFKQE-AVGNSLFASA-N Glu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N IVGJYOOGJLFKQE-AVGNSLFASA-N 0.000 description 1
- IOUQWHIEQYQVFD-JYJNAYRXSA-N Glu-Leu-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IOUQWHIEQYQVFD-JYJNAYRXSA-N 0.000 description 1
- GJBUAAAIZSRCDC-GVXVVHGQSA-N Glu-Leu-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O GJBUAAAIZSRCDC-GVXVVHGQSA-N 0.000 description 1
- YKBUCXNNBYZYAY-MNXVOIDGSA-N Glu-Lys-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YKBUCXNNBYZYAY-MNXVOIDGSA-N 0.000 description 1
- JDUKCSSHWNIQQZ-IHRRRGAJSA-N Glu-Phe-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JDUKCSSHWNIQQZ-IHRRRGAJSA-N 0.000 description 1
- ITVBKCZZLJUUHI-HTUGSXCWSA-N Glu-Phe-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ITVBKCZZLJUUHI-HTUGSXCWSA-N 0.000 description 1
- MIIGESVJEBDJMP-FHWLQOOXSA-N Glu-Phe-Tyr Chemical compound C([C@H](NC(=O)[C@H](CCC(O)=O)N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 MIIGESVJEBDJMP-FHWLQOOXSA-N 0.000 description 1
- SWDNPSMMEWRNOH-HJGDQZAQSA-N Glu-Pro-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O SWDNPSMMEWRNOH-HJGDQZAQSA-N 0.000 description 1
- CAQXJMUDOLSBPF-SUSMZKCASA-N Glu-Thr-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CAQXJMUDOLSBPF-SUSMZKCASA-N 0.000 description 1
- XOEKMEAOMXMURD-JYJNAYRXSA-N Glu-Tyr-His Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O XOEKMEAOMXMURD-JYJNAYRXSA-N 0.000 description 1
- FGGKGJHCVMYGCD-UKJIMTQDSA-N Glu-Val-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FGGKGJHCVMYGCD-UKJIMTQDSA-N 0.000 description 1
- VIPDPMHGICREIS-GVXVVHGQSA-N Glu-Val-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O VIPDPMHGICREIS-GVXVVHGQSA-N 0.000 description 1
- FKJQNJCQTKUBCD-XPUUQOCRSA-N Gly-Ala-His Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)O FKJQNJCQTKUBCD-XPUUQOCRSA-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
- JRDYDYXZKFNNRQ-XPUUQOCRSA-N Gly-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN JRDYDYXZKFNNRQ-XPUUQOCRSA-N 0.000 description 1
- UPOJUWHGMDJUQZ-IUCAKERBSA-N Gly-Arg-Arg Chemical compound NC(=N)NCCC[C@H](NC(=O)CN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O UPOJUWHGMDJUQZ-IUCAKERBSA-N 0.000 description 1
- KKBWDNZXYLGJEY-UHFFFAOYSA-N Gly-Arg-Pro Natural products NCC(=O)NC(CCNC(=N)N)C(=O)N1CCCC1C(=O)O KKBWDNZXYLGJEY-UHFFFAOYSA-N 0.000 description 1
- UXJHNZODTMHWRD-WHFBIAKZSA-N Gly-Asn-Ala Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O UXJHNZODTMHWRD-WHFBIAKZSA-N 0.000 description 1
- LXXANCRPFBSSKS-IUCAKERBSA-N Gly-Gln-Leu Chemical compound [H]NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LXXANCRPFBSSKS-IUCAKERBSA-N 0.000 description 1
- CUYLIWAAAYJKJH-RYUDHWBXSA-N Gly-Glu-Tyr Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 CUYLIWAAAYJKJH-RYUDHWBXSA-N 0.000 description 1
- CCQOOWAONKGYKQ-BYPYZUCNSA-N Gly-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)CN CCQOOWAONKGYKQ-BYPYZUCNSA-N 0.000 description 1
- XMPXVJIDADUOQB-RCOVLWMOSA-N Gly-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C([O-])=O)NC(=O)CNC(=O)C[NH3+] XMPXVJIDADUOQB-RCOVLWMOSA-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
- MVORZMQFXBLMHM-QWRGUYRKSA-N Gly-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 MVORZMQFXBLMHM-QWRGUYRKSA-N 0.000 description 1
- ALOBJFDJTMQQPW-ONGXEEELSA-N Gly-His-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)CN ALOBJFDJTMQQPW-ONGXEEELSA-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
- MIIVFRCYJABHTQ-ONGXEEELSA-N Gly-Leu-Val Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O MIIVFRCYJABHTQ-ONGXEEELSA-N 0.000 description 1
- VBOBNHSVQKKTOT-YUMQZZPRSA-N Gly-Lys-Ala Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O VBOBNHSVQKKTOT-YUMQZZPRSA-N 0.000 description 1
- OQQKUTVULYLCDG-ONGXEEELSA-N Gly-Lys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)CN)C(O)=O OQQKUTVULYLCDG-ONGXEEELSA-N 0.000 description 1
- WZSHYFGOLPXPLL-RYUDHWBXSA-N Gly-Phe-Glu Chemical compound NCC(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CCC(O)=O)C(O)=O WZSHYFGOLPXPLL-RYUDHWBXSA-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
- VNNRLUNBJSWZPF-ZKWXMUAHSA-N Gly-Ser-Ile Chemical compound [H]NCC(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VNNRLUNBJSWZPF-ZKWXMUAHSA-N 0.000 description 1
- ZLCLYFGMKFCDCN-XPUUQOCRSA-N Gly-Ser-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CO)NC(=O)CN)C(O)=O ZLCLYFGMKFCDCN-XPUUQOCRSA-N 0.000 description 1
- XHVONGZZVUUORG-WEDXCCLWSA-N Gly-Thr-Lys Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CCCCN XHVONGZZVUUORG-WEDXCCLWSA-N 0.000 description 1
- 229930186217 Glycolipid Natural products 0.000 description 1
- SVHKVHBPTOMLTO-DCAQKATOSA-N His-Arg-Asp Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O SVHKVHBPTOMLTO-DCAQKATOSA-N 0.000 description 1
- PGTISAJTWZPFGN-PEXQALLHSA-N His-Gly-Ile Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O PGTISAJTWZPFGN-PEXQALLHSA-N 0.000 description 1
- SYIPVNMWBZXKMU-HJPIBITLSA-N His-His-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CC2=CN=CN2)N SYIPVNMWBZXKMU-HJPIBITLSA-N 0.000 description 1
- TWROVBNEHJSXDG-IHRRRGAJSA-N His-Leu-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O TWROVBNEHJSXDG-IHRRRGAJSA-N 0.000 description 1
- IGBBXBFSLKRHJB-BZSNNMDCSA-N His-Lys-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CN=CN1 IGBBXBFSLKRHJB-BZSNNMDCSA-N 0.000 description 1
- KYFGGRHWLFZXPU-KKUMJFAQSA-N His-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N KYFGGRHWLFZXPU-KKUMJFAQSA-N 0.000 description 1
- FBVHRDXSCYELMI-PBCZWWQYSA-N His-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N)O FBVHRDXSCYELMI-PBCZWWQYSA-N 0.000 description 1
- AQCUAZTZSPQJFF-ZKWXMUAHSA-N Ile-Ala-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O AQCUAZTZSPQJFF-ZKWXMUAHSA-N 0.000 description 1
- QYZYJFXHXYUZMZ-UGYAYLCHSA-N Ile-Asn-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N QYZYJFXHXYUZMZ-UGYAYLCHSA-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
- 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
- PNDMHTTXXPUQJH-RWRJDSDZSA-N Ile-Glu-Thr Chemical compound N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H]([C@H](O)C)C(=O)O PNDMHTTXXPUQJH-RWRJDSDZSA-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
- 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
- PHRWFSFCNJPWRO-PPCPHDFISA-N Ile-Leu-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N PHRWFSFCNJPWRO-PPCPHDFISA-N 0.000 description 1
- OVDKXUDMKXAZIV-ZPFDUUQYSA-N Ile-Lys-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)N)C(=O)O)N OVDKXUDMKXAZIV-ZPFDUUQYSA-N 0.000 description 1
- YSGBJIQXTIVBHZ-AJNGGQMLSA-N Ile-Lys-Leu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O YSGBJIQXTIVBHZ-AJNGGQMLSA-N 0.000 description 1
- CKRFDMPBSWYOBT-PPCPHDFISA-N Ile-Lys-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N CKRFDMPBSWYOBT-PPCPHDFISA-N 0.000 description 1
- IMRKCLXPYOIHIF-ZPFDUUQYSA-N Ile-Met-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N IMRKCLXPYOIHIF-ZPFDUUQYSA-N 0.000 description 1
- BATWGBRIZANGPN-ZPFDUUQYSA-N Ile-Pro-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(=O)N)C(=O)O)N BATWGBRIZANGPN-ZPFDUUQYSA-N 0.000 description 1
- CIJLNXXMDUOFPH-HJWJTTGWSA-N Ile-Pro-Phe Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 CIJLNXXMDUOFPH-HJWJTTGWSA-N 0.000 description 1
- JODPUDMBQBIWCK-GHCJXIJMSA-N Ile-Ser-Asn Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O JODPUDMBQBIWCK-GHCJXIJMSA-N 0.000 description 1
- KBDIBHQICWDGDL-PPCPHDFISA-N Ile-Thr-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N KBDIBHQICWDGDL-PPCPHDFISA-N 0.000 description 1
- YJRSIJZUIUANHO-NAKRPEOUSA-N Ile-Val-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(=O)O)N YJRSIJZUIUANHO-NAKRPEOUSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 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
- LHSGPCFBGJHPCY-UHFFFAOYSA-N L-leucine-L-tyrosine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 LHSGPCFBGJHPCY-UHFFFAOYSA-N 0.000 description 1
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 1
- KWTVLKBOQATPHJ-SRVKXCTJSA-N Leu-Ala-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(C)C)N KWTVLKBOQATPHJ-SRVKXCTJSA-N 0.000 description 1
- ILJREDZFPHTUIE-GUBZILKMSA-N Leu-Asp-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ILJREDZFPHTUIE-GUBZILKMSA-N 0.000 description 1
- DLCXCECTCPKKCD-GUBZILKMSA-N Leu-Gln-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O DLCXCECTCPKKCD-GUBZILKMSA-N 0.000 description 1
- DPWGZWUMUUJQDT-IUCAKERBSA-N Leu-Gln-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O DPWGZWUMUUJQDT-IUCAKERBSA-N 0.000 description 1
- DZQMXBALGUHGJT-GUBZILKMSA-N Leu-Glu-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O DZQMXBALGUHGJT-GUBZILKMSA-N 0.000 description 1
- OXRLYTYUXAQTHP-YUMQZZPRSA-N Leu-Gly-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(O)=O OXRLYTYUXAQTHP-YUMQZZPRSA-N 0.000 description 1
- KGCLIYGPQXUNLO-IUCAKERBSA-N Leu-Gly-Glu Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O KGCLIYGPQXUNLO-IUCAKERBSA-N 0.000 description 1
- KVOFSTUWVSQMDK-KKUMJFAQSA-N Leu-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(C)C)CC1=CN=CN1 KVOFSTUWVSQMDK-KKUMJFAQSA-N 0.000 description 1
- KOSWSHVQIVTVQF-ZPFDUUQYSA-N Leu-Ile-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O KOSWSHVQIVTVQF-ZPFDUUQYSA-N 0.000 description 1
- IFMPDNRWZZEZSL-SRVKXCTJSA-N Leu-Leu-Cys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(O)=O IFMPDNRWZZEZSL-SRVKXCTJSA-N 0.000 description 1
- ZRHDPZAAWLXXIR-SRVKXCTJSA-N Leu-Lys-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O ZRHDPZAAWLXXIR-SRVKXCTJSA-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
- FIICHHJDINDXKG-IHPCNDPISA-N Leu-Lys-Trp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O FIICHHJDINDXKG-IHPCNDPISA-N 0.000 description 1
- FYPWFNKQVVEELI-ULQDDVLXSA-N Leu-Phe-Val Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C(C)C)C(O)=O)CC1=CC=CC=C1 FYPWFNKQVVEELI-ULQDDVLXSA-N 0.000 description 1
- VULJUQZPSOASBZ-SRVKXCTJSA-N Leu-Pro-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O VULJUQZPSOASBZ-SRVKXCTJSA-N 0.000 description 1
- ZJZNLRVCZWUONM-JXUBOQSCSA-N Leu-Thr-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O ZJZNLRVCZWUONM-JXUBOQSCSA-N 0.000 description 1
- FBNPMTNBFFAMMH-UHFFFAOYSA-N Leu-Val-Arg Natural products CC(C)CC(N)C(=O)NC(C(C)C)C(=O)NC(C(O)=O)CCCN=C(N)N FBNPMTNBFFAMMH-UHFFFAOYSA-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
- FZIJIFCXUCZHOL-CIUDSAMLSA-N Lys-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN FZIJIFCXUCZHOL-CIUDSAMLSA-N 0.000 description 1
- MPOHDJKRBLVGCT-CIUDSAMLSA-N Lys-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCCN)N MPOHDJKRBLVGCT-CIUDSAMLSA-N 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
- GGNOBVSOZPHLCE-GUBZILKMSA-N Lys-Gln-Asp Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O GGNOBVSOZPHLCE-GUBZILKMSA-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
- DCRWPTBMWMGADO-AVGNSLFASA-N Lys-Glu-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O DCRWPTBMWMGADO-AVGNSLFASA-N 0.000 description 1
- IMAKMJCBYCSMHM-AVGNSLFASA-N Lys-Glu-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN IMAKMJCBYCSMHM-AVGNSLFASA-N 0.000 description 1
- OWRUUFUVXFREBD-KKUMJFAQSA-N Lys-His-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(O)=O OWRUUFUVXFREBD-KKUMJFAQSA-N 0.000 description 1
- MYZMQWHPDAYKIE-SRVKXCTJSA-N Lys-Leu-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O MYZMQWHPDAYKIE-SRVKXCTJSA-N 0.000 description 1
- VMTYLUGCXIEDMV-QWRGUYRKSA-N Lys-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCCCN VMTYLUGCXIEDMV-QWRGUYRKSA-N 0.000 description 1
- QQPSCXKFDSORFT-IHRRRGAJSA-N Lys-Lys-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCCN QQPSCXKFDSORFT-IHRRRGAJSA-N 0.000 description 1
- ALEVUGKHINJNIF-QEJZJMRPSA-N Lys-Phe-Ala Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=CC=C1 ALEVUGKHINJNIF-QEJZJMRPSA-N 0.000 description 1
- AEIIJFBQVGYVEV-YESZJQIVSA-N Lys-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CCCCN)N)C(=O)O AEIIJFBQVGYVEV-YESZJQIVSA-N 0.000 description 1
- AFLBTVGQCQLOFJ-AVGNSLFASA-N Lys-Pro-Arg Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O AFLBTVGQCQLOFJ-AVGNSLFASA-N 0.000 description 1
- QBHGXFQJFPWJIH-XUXIUFHCSA-N Lys-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCCCN QBHGXFQJFPWJIH-XUXIUFHCSA-N 0.000 description 1
- MIROMRNASYKZNL-ULQDDVLXSA-N Lys-Pro-Tyr Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 MIROMRNASYKZNL-ULQDDVLXSA-N 0.000 description 1
- DNWBUCHHMRQWCZ-GUBZILKMSA-N Lys-Ser-Gln Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(N)=O DNWBUCHHMRQWCZ-GUBZILKMSA-N 0.000 description 1
- UWHCKWNPWKTMBM-WDCWCFNPSA-N Lys-Thr-Gln Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O UWHCKWNPWKTMBM-WDCWCFNPSA-N 0.000 description 1
- FPQMQEOVSKMVMA-ACRUOGEOSA-N Lys-Tyr-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)NC(=O)[C@H](CCCCN)N)O FPQMQEOVSKMVMA-ACRUOGEOSA-N 0.000 description 1
- UGCIQUYEJIEHKX-GVXVVHGQSA-N Lys-Val-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O UGCIQUYEJIEHKX-GVXVVHGQSA-N 0.000 description 1
- VWJFOUBDZIUXGA-AVGNSLFASA-N Lys-Val-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CCCCN)N VWJFOUBDZIUXGA-AVGNSLFASA-N 0.000 description 1
- NSGXXVIHCIAISP-CIUDSAMLSA-N Met-Asn-Gln Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O NSGXXVIHCIAISP-CIUDSAMLSA-N 0.000 description 1
- BQVJARUIXRXDKN-DCAQKATOSA-N Met-Asn-His Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 BQVJARUIXRXDKN-DCAQKATOSA-N 0.000 description 1
- KQBJYJXPZBNEIK-DCAQKATOSA-N Met-Glu-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KQBJYJXPZBNEIK-DCAQKATOSA-N 0.000 description 1
- SJDQOYTYNGZZJX-SRVKXCTJSA-N Met-Glu-Leu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O SJDQOYTYNGZZJX-SRVKXCTJSA-N 0.000 description 1
- HZLSUXCMSIBCRV-RVMXOQNASA-N Met-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCSC)N HZLSUXCMSIBCRV-RVMXOQNASA-N 0.000 description 1
- UROWNMBTQGGTHB-DCAQKATOSA-N Met-Leu-Asp Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O UROWNMBTQGGTHB-DCAQKATOSA-N 0.000 description 1
- LLKWSEXLNFBKIF-CYDGBPFRSA-N Met-Met-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCSC)NC(=O)[C@@H](N)CCSC LLKWSEXLNFBKIF-CYDGBPFRSA-N 0.000 description 1
- CIDICGYKRUTYLE-FXQIFTODSA-N Met-Ser-Ala Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O CIDICGYKRUTYLE-FXQIFTODSA-N 0.000 description 1
- CQRGINSEMFBACV-WPRPVWTQSA-N Met-Val-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O CQRGINSEMFBACV-WPRPVWTQSA-N 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-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
- 108010065395 Neuropep-1 Proteins 0.000 description 1
- 206010067482 No adverse event Diseases 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- BBDSZDHUCPSYAC-QEJZJMRPSA-N Phe-Ala-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O BBDSZDHUCPSYAC-QEJZJMRPSA-N 0.000 description 1
- YYRCPTVAPLQRNC-ULQDDVLXSA-N Phe-Arg-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CC1=CC=CC=C1 YYRCPTVAPLQRNC-ULQDDVLXSA-N 0.000 description 1
- FRPVPGRXUKFEQE-YDHLFZDLSA-N Phe-Asp-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O FRPVPGRXUKFEQE-YDHLFZDLSA-N 0.000 description 1
- LLGTYVHITPVGKR-RYUDHWBXSA-N Phe-Gln-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O LLGTYVHITPVGKR-RYUDHWBXSA-N 0.000 description 1
- UAMFZRNCIFFMLE-FHWLQOOXSA-N Phe-Glu-Tyr Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N UAMFZRNCIFFMLE-FHWLQOOXSA-N 0.000 description 1
- NAXPHWZXEXNDIW-JTQLQIEISA-N Phe-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 NAXPHWZXEXNDIW-JTQLQIEISA-N 0.000 description 1
- DZVXMMSUWWUIQE-ACRUOGEOSA-N Phe-His-Tyr Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)O)N DZVXMMSUWWUIQE-ACRUOGEOSA-N 0.000 description 1
- WEMYTDDMDBLPMI-DKIMLUQUSA-N Phe-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N WEMYTDDMDBLPMI-DKIMLUQUSA-N 0.000 description 1
- BYAIIACBWBOJCU-URLPEUOOSA-N Phe-Ile-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O BYAIIACBWBOJCU-URLPEUOOSA-N 0.000 description 1
- YTILBRIUASDGBL-BZSNNMDCSA-N Phe-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 YTILBRIUASDGBL-BZSNNMDCSA-N 0.000 description 1
- TXJJXEXCZBHDNA-ACRUOGEOSA-N Phe-Phe-His Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N[C@@H](CC3=CN=CN3)C(=O)O)N TXJJXEXCZBHDNA-ACRUOGEOSA-N 0.000 description 1
- QARPMYDMYVLFMW-KKUMJFAQSA-N Phe-Pro-Glu Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(O)=O)C1=CC=CC=C1 QARPMYDMYVLFMW-KKUMJFAQSA-N 0.000 description 1
- BSKMOCNNLNDIMU-CDMKHQONSA-N Phe-Thr-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O BSKMOCNNLNDIMU-CDMKHQONSA-N 0.000 description 1
- MMPBPRXOFJNCCN-ZEWNOJEFSA-N Phe-Tyr-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MMPBPRXOFJNCCN-ZEWNOJEFSA-N 0.000 description 1
- DZZCICYRSZASNF-FXQIFTODSA-N Pro-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 DZZCICYRSZASNF-FXQIFTODSA-N 0.000 description 1
- UTAUEDINXUMHLG-FXQIFTODSA-N Pro-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@@H]1CCCN1 UTAUEDINXUMHLG-FXQIFTODSA-N 0.000 description 1
- ILMLVTGTUJPQFP-FXQIFTODSA-N Pro-Asp-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O ILMLVTGTUJPQFP-FXQIFTODSA-N 0.000 description 1
- YFNOUBWUIIJQHF-LPEHRKFASA-N Pro-Asp-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC(=O)O)C(=O)N2CCC[C@@H]2C(=O)O YFNOUBWUIIJQHF-LPEHRKFASA-N 0.000 description 1
- CLNJSLSHKJECME-BQBZGAKWSA-N Pro-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H]1CCCN1 CLNJSLSHKJECME-BQBZGAKWSA-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
- KWMUAKQOVYCQJQ-ZPFDUUQYSA-N Pro-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@@H]1CCCN1 KWMUAKQOVYCQJQ-ZPFDUUQYSA-N 0.000 description 1
- SMFQZMGHCODUPQ-ULQDDVLXSA-N Pro-Lys-Phe Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SMFQZMGHCODUPQ-ULQDDVLXSA-N 0.000 description 1
- BLJMJZOMZRCESA-GUBZILKMSA-N Pro-Met-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@@H]1CCCN1 BLJMJZOMZRCESA-GUBZILKMSA-N 0.000 description 1
- GBUNEGKQPSAMNK-QTKMDUPCSA-N Pro-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@@H]2CCCN2)O GBUNEGKQPSAMNK-QTKMDUPCSA-N 0.000 description 1
- FDMCIBSQRKFSTJ-RHYQMDGZSA-N Pro-Thr-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O FDMCIBSQRKFSTJ-RHYQMDGZSA-N 0.000 description 1
- CNUIHOAISPKQPY-HSHDSVGOSA-N Pro-Thr-Trp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O CNUIHOAISPKQPY-HSHDSVGOSA-N 0.000 description 1
- LEBTWGWVUVJNTA-FKBYEOEOSA-N Pro-Trp-Phe Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)N[C@@H](CC4=CC=CC=C4)C(=O)O LEBTWGWVUVJNTA-FKBYEOEOSA-N 0.000 description 1
- 101710194807 Protective antigen Proteins 0.000 description 1
- 101800000460 Protein TF Proteins 0.000 description 1
- 101000933967 Pseudomonas phage KPP25 Major capsid protein 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
- FCRMLGJMPXCAHD-FXQIFTODSA-N Ser-Arg-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O FCRMLGJMPXCAHD-FXQIFTODSA-N 0.000 description 1
- QGMLKFGTGXWAHF-IHRRRGAJSA-N Ser-Arg-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QGMLKFGTGXWAHF-IHRRRGAJSA-N 0.000 description 1
- QPFJSHSJFIYDJZ-GHCJXIJMSA-N Ser-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CO QPFJSHSJFIYDJZ-GHCJXIJMSA-N 0.000 description 1
- VMVNCJDKFOQOHM-GUBZILKMSA-N Ser-Gln-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CO)N VMVNCJDKFOQOHM-GUBZILKMSA-N 0.000 description 1
- YMAWDPHQVABADW-CIUDSAMLSA-N Ser-Gln-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O YMAWDPHQVABADW-CIUDSAMLSA-N 0.000 description 1
- BRGQQXQKPUCUJQ-KBIXCLLPSA-N Ser-Glu-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BRGQQXQKPUCUJQ-KBIXCLLPSA-N 0.000 description 1
- KDGARKCAKHBEDB-NKWVEPMBSA-N Ser-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CO)N)C(=O)O KDGARKCAKHBEDB-NKWVEPMBSA-N 0.000 description 1
- HDBOEVPDIDDEPC-CIUDSAMLSA-N Ser-Lys-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O HDBOEVPDIDDEPC-CIUDSAMLSA-N 0.000 description 1
- WGDYNRCOQRERLZ-KKUMJFAQSA-N Ser-Lys-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)N WGDYNRCOQRERLZ-KKUMJFAQSA-N 0.000 description 1
- PTWIYDNFWPXQSD-GARJFASQSA-N Ser-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CO)N)C(=O)O PTWIYDNFWPXQSD-GARJFASQSA-N 0.000 description 1
- FZEUTKVQGMVGHW-AVGNSLFASA-N Ser-Phe-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O FZEUTKVQGMVGHW-AVGNSLFASA-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
- JCLAFVNDBJMLBC-JBDRJPRFSA-N Ser-Ser-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JCLAFVNDBJMLBC-JBDRJPRFSA-N 0.000 description 1
- UQGAAZXSCGWMFU-UBHSHLNASA-N Ser-Trp-Asp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CO)N UQGAAZXSCGWMFU-UBHSHLNASA-N 0.000 description 1
- FZNNGIHSIPKFRE-QEJZJMRPSA-N Ser-Trp-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(N)=O)C(O)=O FZNNGIHSIPKFRE-QEJZJMRPSA-N 0.000 description 1
- JGUWRQWULDWNCM-FXQIFTODSA-N Ser-Val-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O JGUWRQWULDWNCM-FXQIFTODSA-N 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 101150025711 TF gene Proteins 0.000 description 1
- 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 1
- BSNZTJXVDOINSR-JXUBOQSCSA-N Thr-Ala-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O BSNZTJXVDOINSR-JXUBOQSCSA-N 0.000 description 1
- QGXCWPNQVCYJEL-NUMRIWBASA-N Thr-Asn-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O QGXCWPNQVCYJEL-NUMRIWBASA-N 0.000 description 1
- DCLBXIWHLVEPMQ-JRQIVUDYSA-N Thr-Asp-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 DCLBXIWHLVEPMQ-JRQIVUDYSA-N 0.000 description 1
- MMTOHPRBJKEZHT-BWBBJGPYSA-N Thr-Cys-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(O)=O MMTOHPRBJKEZHT-BWBBJGPYSA-N 0.000 description 1
- QILPDQCTQZDHFM-HJGDQZAQSA-N Thr-Gln-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QILPDQCTQZDHFM-HJGDQZAQSA-N 0.000 description 1
- UHBPFYOQQPFKQR-JHEQGTHGSA-N Thr-Gln-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O UHBPFYOQQPFKQR-JHEQGTHGSA-N 0.000 description 1
- HJOSVGCWOTYJFG-WDCWCFNPSA-N Thr-Glu-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N)O HJOSVGCWOTYJFG-WDCWCFNPSA-N 0.000 description 1
- 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 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
- XOWKUMFHEZLKLT-CIQUZCHMSA-N Thr-Ile-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O XOWKUMFHEZLKLT-CIQUZCHMSA-N 0.000 description 1
- CRZNCABIJLRFKZ-IUKAMOBKSA-N Thr-Ile-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N CRZNCABIJLRFKZ-IUKAMOBKSA-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
- 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
- BIBYEFRASCNLAA-CDMKHQONSA-N Thr-Phe-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=CC=C1 BIBYEFRASCNLAA-CDMKHQONSA-N 0.000 description 1
- XKWABWFMQXMUMT-HJGDQZAQSA-N Thr-Pro-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O XKWABWFMQXMUMT-HJGDQZAQSA-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
- DEGCBBCMYWNJNA-RHYQMDGZSA-N Thr-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)[C@@H](C)O DEGCBBCMYWNJNA-RHYQMDGZSA-N 0.000 description 1
- 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 1
- JAWUQFCGNVEDRN-MEYUZBJRSA-N Thr-Tyr-Leu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N)O JAWUQFCGNVEDRN-MEYUZBJRSA-N 0.000 description 1
- DIHPMRTXPYMDJZ-KAOXEZKKSA-N Thr-Tyr-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N2CCC[C@@H]2C(=O)O)N)O DIHPMRTXPYMDJZ-KAOXEZKKSA-N 0.000 description 1
- CYCGARJWIQWPQM-YJRXYDGGSA-N Thr-Tyr-Ser Chemical compound C[C@@H](O)[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CO)C([O-])=O)CC1=CC=C(O)C=C1 CYCGARJWIQWPQM-YJRXYDGGSA-N 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- FGJWNBBFAUHBEP-IHPCNDPISA-N Tyr-Asp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N FGJWNBBFAUHBEP-IHPCNDPISA-N 0.000 description 1
- AKLNEFNQWLHIGY-QWRGUYRKSA-N Tyr-Gly-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)O)C(=O)O)N)O AKLNEFNQWLHIGY-QWRGUYRKSA-N 0.000 description 1
- JKUZFODWJGEQAP-KBPBESRZSA-N Tyr-Gly-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)O)N)O JKUZFODWJGEQAP-KBPBESRZSA-N 0.000 description 1
- OHOVFPKXPZODHS-SJWGOKEGSA-N Tyr-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N OHOVFPKXPZODHS-SJWGOKEGSA-N 0.000 description 1
- FWOVTJKVUCGVND-UFYCRDLUSA-N Tyr-Met-Phe Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N FWOVTJKVUCGVND-UFYCRDLUSA-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
- KWKJGBHDYJOVCR-SRVKXCTJSA-N Tyr-Ser-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N)O KWKJGBHDYJOVCR-SRVKXCTJSA-N 0.000 description 1
- UMSZZGTXGKHTFJ-SRVKXCTJSA-N Tyr-Ser-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 UMSZZGTXGKHTFJ-SRVKXCTJSA-N 0.000 description 1
- LUMQYLVYUIRHHU-YJRXYDGGSA-N Tyr-Ser-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LUMQYLVYUIRHHU-YJRXYDGGSA-N 0.000 description 1
- OBKOPLHSRDATFO-XHSDSOJGSA-N Tyr-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N OBKOPLHSRDATFO-XHSDSOJGSA-N 0.000 description 1
- JIODCDXKCJRMEH-NHCYSSNCSA-N Val-Arg-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N JIODCDXKCJRMEH-NHCYSSNCSA-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
- QPZMOUMNTGTEFR-ZKWXMUAHSA-N Val-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C(C)C)N QPZMOUMNTGTEFR-ZKWXMUAHSA-N 0.000 description 1
- OGNMURQZFMHFFD-NHCYSSNCSA-N Val-Asn-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N OGNMURQZFMHFFD-NHCYSSNCSA-N 0.000 description 1
- QHDXUYOYTPWCSK-RCOVLWMOSA-N Val-Asp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)NCC(=O)O)N QHDXUYOYTPWCSK-RCOVLWMOSA-N 0.000 description 1
- GBESYURLQOYWLU-LAEOZQHASA-N Val-Glu-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N GBESYURLQOYWLU-LAEOZQHASA-N 0.000 description 1
- XWYUBUYQMOUFRQ-IFFSRLJSSA-N Val-Glu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](C(C)C)N)O XWYUBUYQMOUFRQ-IFFSRLJSSA-N 0.000 description 1
- UEHRGZCNLSWGHK-DLOVCJGASA-N Val-Glu-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O UEHRGZCNLSWGHK-DLOVCJGASA-N 0.000 description 1
- BEGDZYNDCNEGJZ-XVKPBYJWSA-N Val-Gly-Gln Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(N)=O BEGDZYNDCNEGJZ-XVKPBYJWSA-N 0.000 description 1
- KDKLLPMFFGYQJD-CYDGBPFRSA-N Val-Ile-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](C(C)C)N KDKLLPMFFGYQJD-CYDGBPFRSA-N 0.000 description 1
- BZMIYHIJVVJPCK-QSFUFRPTSA-N Val-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N BZMIYHIJVVJPCK-QSFUFRPTSA-N 0.000 description 1
- OTJMMKPMLUNTQT-AVGNSLFASA-N Val-Leu-Arg Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](C(C)C)N OTJMMKPMLUNTQT-AVGNSLFASA-N 0.000 description 1
- UMPVMAYCLYMYGA-ONGXEEELSA-N Val-Leu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O UMPVMAYCLYMYGA-ONGXEEELSA-N 0.000 description 1
- JAKHAONCJJZVHT-DCAQKATOSA-N Val-Lys-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)O)N JAKHAONCJJZVHT-DCAQKATOSA-N 0.000 description 1
- QSPOLEBZTMESFY-SRVKXCTJSA-N Val-Pro-Val Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O QSPOLEBZTMESFY-SRVKXCTJSA-N 0.000 description 1
- DEGUERSKQBRZMZ-FXQIFTODSA-N Val-Ser-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O DEGUERSKQBRZMZ-FXQIFTODSA-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
- NZYNRRGJJVSSTJ-GUBZILKMSA-N Val-Ser-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O NZYNRRGJJVSSTJ-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
- TVGWMCTYUFBXAP-QTKMDUPCSA-N Val-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C(C)C)N)O TVGWMCTYUFBXAP-QTKMDUPCSA-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
- PMKQKNBISAOSRI-XHSDSOJGSA-N Val-Tyr-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N2CCC[C@@H]2C(=O)O)N PMKQKNBISAOSRI-XHSDSOJGSA-N 0.000 description 1
- OWFGFHQMSBTKLX-UFYCRDLUSA-N Val-Tyr-Tyr Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N OWFGFHQMSBTKLX-UFYCRDLUSA-N 0.000 description 1
- 108010067390 Viral Proteins Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010052670 arginyl-glutamyl-glutamic acid Proteins 0.000 description 1
- 108010007483 arginyl-leucyl-tyrosyl-glutamic acid Proteins 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 108010010430 asparagine-proline-alanine Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 229940031567 attenuated vaccine Drugs 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 210000000234 capsid Anatomy 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000013530 defoamer Substances 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 238000011013 endotoxin removal Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000006052 feed supplement Substances 0.000 description 1
- 230000035611 feeding Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010042598 glutamyl-aspartyl-glycine 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
- 108010075431 glycyl-alanyl-phenylalanine Proteins 0.000 description 1
- 108010027668 glycyl-alanyl-valine Proteins 0.000 description 1
- 108010038983 glycyl-histidyl-lysine Proteins 0.000 description 1
- 108010082286 glycyl-seryl-alanine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000021760 high fever Diseases 0.000 description 1
- 108010040030 histidinoalanine Proteins 0.000 description 1
- 108010028295 histidylhistidine Proteins 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 229940031551 inactivated vaccine Drugs 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 108010053037 kyotorphin Proteins 0.000 description 1
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 1
- 108010030617 leucyl-phenylalanyl-valine Proteins 0.000 description 1
- 108010000761 leucylarginine Proteins 0.000 description 1
- 108010012058 leucyltyrosine Proteins 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 108010005942 methionylglycine Proteins 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- -1 pH 9.5) Substances 0.000 description 1
- 108010070409 phenylalanyl-glycyl-glycine Proteins 0.000 description 1
- 108010089198 phenylalanyl-prolyl-arginine Proteins 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 108010025488 pinealon Proteins 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000003761 preservation solution Substances 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108010004914 prolylarginine Proteins 0.000 description 1
- 108010070643 prolylglutamic acid Proteins 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 238000011076 safety test Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000012089 stop solution Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 108010033670 threonyl-aspartyl-tyrosine Proteins 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 108700004896 tripeptide FEG Proteins 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 108010084932 tryptophyl-proline Proteins 0.000 description 1
- 108010020532 tyrosyl-proline Proteins 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000008478 viral entry into host cell Effects 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 210000001835 viscera Anatomy 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
- A61K39/12—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
- A61P31/20—Antivirals for DNA 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
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- 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
- 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
- C12N2710/12011—Asfarviridae
- C12N2710/12022—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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/12011—Asfarviridae
- C12N2710/12034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Hematology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Urology & Nephrology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Zoology (AREA)
- Cell Biology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Food Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Communicable Diseases (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Plant Pathology (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Mycology (AREA)
Abstract
本发明公开了一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白及其制备方法和应用，所述非洲猪瘟病毒p72亚单位可溶性融合蛋白包括OPTI‑p72蛋白和一种伴侣蛋白，其中所述OPTI‑p72蛋白的核苷酸序列为经优化后的序列OPTI‑p72如SEQ ID NO.1所示。本发明能够在大肠杆菌中直接可溶性表达P72，克服了现有技术中的诸多问题，且制备方法简单、成本低。
Description
技术领域
本发明属于兽用生物制品技术领域。涉及一种重组的非洲猪瘟P72亚单位可溶性融合蛋白及在制备诊断、预防和治疗非洲猪瘟疫苗中的应用。
背景技术
非洲猪瘟(African swine fever，ASF)是由非洲猪瘟病毒(African swine fevervirus，ASFV)引起的猪的急性、热性、高度接触性传染病。猪是ASFV自然感染的唯一哺乳动物宿主，包括家猪和野猪，尤其是家猪，易感性极高。猪感染非洲猪瘟病毒后临床上以皮肤充血，内脏器官出血，高热为特征，发病率及死亡率高达100％。世界动物卫生组织将其列为A类疫病，我国也将其列为一类动物传染病。
2018年8月份以来，非洲猪瘟在我国多个省份爆发，给我国养猪业带来了严重的经济损失。尽管已经有许多非洲猪瘟的研究性报告，但目前世界上尚未有有效预防非洲猪瘟的疫苗和治疗该病的药物发现，迫切需要新型疫苗的研发和生产来预防非洲猪瘟。
ASFV病毒是具有囊膜的虫媒DNA病毒。病毒颗粒呈二十面体对称结构，平均直径200nm，表面有含糖脂类的囊膜覆盖。其病毒基因组为双股线性DNA，大小为170-190kb，整个基因组大约有150个ORF，编码150-200个蛋白质。P72蛋白由ASFV病毒B646L基因编码的，分子量大小约32kD的蛋白。是非洲猪瘟病毒粒子形成二十面体衣壳的的主要结构蛋白，约占整个病毒粒子的32％。研究发现，p72蛋白相对比较保守，有稳定的抗原性。且与病毒进入宿主细胞的过程有关，针对p72的抗体能够抑制病毒与巨噬细胞的结合。因此，p72是一个很好的保护性抗原。目前，已经有P72蛋白能成功的表达在原核系统，但表达产量低，且以包涵体形式存在(具体参见公开号为CN103805615的发明专利)，无法满足基因工程亚单位疫苗的开发。而利用本发明提供的方法可大量可溶性表达P72蛋白。在当前没有可能大规模制备灭活疫苗或弱毒疫苗的情况下，确定一种制备该病毒的免疫原性蛋白的方法，以便研究一种能够预防该疾病的亚单位疫苗或具有能够诊断该疾病的试剂具有重大的意义。
发明内容
在现有技术的基础上，本发明为了克服现有技术中存在的诸多缺陷(如表达产量低，包涵体表达，纯化困难成本相对较高等)，提供了一种制作成本低廉可在大肠杆菌中直接可溶性表达p72亚单位可溶性融合蛋白及其制备方法。
根据本发明的一个方面，本发明提供了一种可在大肠杆菌中表达p72蛋白的优化后的OPTI-p72的核苷酸序列。为了能够在大肠杆菌中高效表达P72蛋白，本发明根据GenBank:FR682468.1上已经存在公开的序列(密码子优化前的编码P72蛋白的核苷酸序列如SEQ ID NO.2所示)，我们对p72基因进行分析，首先对p72基因编码的密码子的在原核表达中的密码子偏好性进行了优化，其次对p72基因的GC含量和mRNA在大肠杆菌的稳定性进行优化。所述最终优化后OPTI-p72的核苷酸序列如SEQ ID NO.1所示。因此，所述非洲猪瘟病毒p72亚单位可溶性融合蛋白包括OPTI-p72蛋白和一种伴侣蛋白，其中所述OPTI-p72蛋白的核苷酸序列为p72蛋白经优化后的序列OPTI-p72如SEQ ID NO.1所示。
在本发明的优选技术方案中，所述伴侣蛋白为TF蛋白。
在本发明的优选技术方案中，优化后的OPTI-p72直接在原核表达系统中也不能可溶性表达p72，为了能够得到可溶性表达P72蛋白，本发明在OPTI-p72核苷酸序列3’端或5’端加入伴侣蛋白的基因编码序列，其中伴侣蛋白优选为TF，得到OPTI-TF-P72。优选地，5’端加入TF基因编码OPTI-TF-p72的基因编码序列如SEQ ID NO.3所示，其氨基酸序列如SEQ IDNO.4所示。
为了方便使用亲和层析的方法纯化融合蛋白，根据本发明的技术方案，优先地，所述非洲猪瘟病毒p30亚单位可溶性融合蛋白在如SEQ ID NO.4所示的氨基酸序列的氨基末端或羧基末端连接有poly-His、FLAG、c-myc、HA和poly-Arg中的一种标签。
根据本发明的另一个方面，为了能够在大肠杆菌中高效可溶性表达P72蛋白，本发明将OPTI-TF-p72-6His核苷酸序列导入原核表达载体。所述表达载体可以是任何一种原核表达载体，所述的表达载体具体但不限于pET30a表达载体、pBAD载体、pcold载体、pQE载体、pKK载体等。优先地，所述的载体为pET30a载体。
根据本发明的另一个方面，本发明提供了一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白的制备方法，所述制备方法包括以下步骤：1)将含有OPTI-p72蛋白和一种伴侣蛋白的基因克隆到原核表达载体中，得到含有非洲猪瘟病毒OPTI-p72蛋白和一种伴侣蛋白编码基因的重组质粒；其中，所述OPTI-p72蛋白的核苷酸序列为p72蛋白经优化后的序列OPTI-p72如SEQ ID NO.1所示；2)再将步骤1)所述重组质粒转化至大肠杆菌菌株的细胞中，得到重组表达的菌株；3)通过培养、筛选、步骤2)中所述重组表达的菌株，得到高度表达的菌株；4)发酵培养步骤3)中所述高度表达的菌株，纯化后得到重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白。
在本发明的优选技术方案中，步骤1)中，所述伴侣蛋白为TF蛋白。
在本发明的优选技术方案中，含有非洲猪瘟病毒OPTI-p72蛋白和TF蛋白的核苷酸序列OPTI-TF-p72是OPTI-p72蛋白的5’端插入TF蛋白的核苷酸序列，所述OPTI-TF-p72蛋白的核苷酸序列如SEQ ID NO.3所示。
在本发明的优选技术方案中，含有非洲猪瘟病毒OPTI-p72蛋白和TF蛋白的氨基酸序列如SEQ ID NO.4所示。
在本发明的优选技术方案中，含有非洲猪瘟病毒OPTI-p72蛋白和TF蛋白在如SEQID NO.4所示的氨基酸序列的氨基末端或羧基末端连接有poly-His、FLAG、c-myc、HA和poly-Arg中的一种标签。
在本发明的优选技术方案中，为了能够在大肠杆菌中高效可溶性表达P72蛋白，本发明将OPTI-TF-p72-6His核苷酸序列导入原核表达载体。所述表达载体可以是任何一种原核表达载体，所述的表达载体具体但不限于pET30a表达载体、pBAD载体、pcold载体、pQE载体、pKK载体等。更优选地，所述载体为pET30a载体。
在本发明的优选技术方案中，步骤2)中，所述大肠杆菌菌株选自BL21(DE3)、BL21star(DE3)、arcticexpress中的一种菌株。更优选地，步骤2)中所述大肠杆菌菌株使用BL21(DE3)。
在本发明的技术方案中，优先地，所述的步骤3)中使用IPTG的浓度为0.2mmol/L。
在本发明的技术方案中，优先地，所述的步骤4)中使用裂解液：20mM Tris(pH8.7)，500mM NaCl，0.2％TritonX-114，5％甘油，1mMβ-ME。
根据本发明的再一个方面，本发明提供了一种所述非洲猪瘟病毒p72亚单位可溶性融合蛋白在制备诊断、预防和治疗非洲猪瘟的疫苗中的应用。
与现有技术相比，本发明公开的表达序列、表达载体以及相应的纯化制备方法克服了现有技术中的缺陷，解决了大肠杆菌中不能直接大量可溶性表达P72蛋白且产量低的问题。本发明能够在大肠杆菌中直接可溶性表达P72，克服了现有技术中的诸多问题，且制备方法简单、成本低。
附图说明
图1 P72蛋白编码的核苷酸优化前后序列比对结果。
图2 pET30-OPTI-TF-p72-6His构建示意图。
图3 pET30-OPTI-TF-p72-6His酶切鉴定结果。1-4：pET30a-OPTI-TF-P72质粒SacI和SalI双酶切，条带大小分别为6673bp，1979bp，质粒酶切正确。M：DL10000Marker
图4 SDS-PAGE检测TF-P72蛋白诱导表达的结果。1：诱导前上清；2：诱导前沉淀；3：诱导后上清；4：诱导后沉淀。M：蛋白Marker 26619。
图5 SDS-PAGE检测TF-P72蛋白纯化后的结果。1：蛋白Marker 26619，2：纯化后的TF-P72蛋白纯度85％。
图6 SDS-PAGE检测第十次取样4℃和-20℃保存的TF-P72蛋白。1是蛋白Marker26619，2，3:第十次样品4℃和-20℃蛋白样品，上样量3ug。
具体实施方式
以下将结合附图和实施例对本发明做进一步说明，本发明的实施例仅用于说明本发明的技术方案，并非限定本发明。
使用试剂均为国产市售产品。
实施例1 P72蛋白表达及制备
1.1非洲猪瘟P72蛋白的选择
非洲猪瘟结构蛋白P72是由B646L基因编码的一段多肽，P72蛋白合成的动力学表明该蛋白在感染晚期被翻译，约占病毒蛋白量32％，是病毒二十面体结构的主要蛋白。因此，利用P72蛋白作为抗原具有很好的防控非洲猪瘟的感染或作为诊断试剂检测。尽管P72蛋白报道在多个系统中存在表达，但是，目前还没有报道在原核表达系统中能可溶性表达及纯化得到该蛋白，这也是本发明所要解决的一个重要的技术问题。
为了使所述亚单位TF-P72蛋白便于纯化，可在如SEQ ID NO.4所示的氨基酸序列的氨基末端或羧基末端连接如表一所示的标签，本实施例中具体以在羧基端添加Poly-His为例，将其连接于如SEQ ID NO.4所示的氨基酸序列的羧基末端处。
表一 标签及其氨基酸序列
1.2非洲猪瘟P72蛋白密码子优化
本实验室以2018年在中国报道流行的非洲猪瘟毒株亚型，参考Georgia 2007/1全基因序列(GenBank：FR682468.1)为模板，对编码非洲猪瘟P72蛋白的B646L的核苷酸序列进行密码子优化，得到OPTI-p72序列，如SEQ ID NO.1所示，该序列合成工作委托南京金斯瑞生物科技有限公司完成。如图1所示，优化前后有24.6％的核苷酸序列不同。
1.3表达载体构建及验证
1.3.1 PCR扩增目的片段
1.3.1.1 PCR反应
(1)引物设计及合成
上游引物:5’-tggagctcGAGAACCTGTACTTCCAGGGTGCGAGCGGTGGCGCG-3’
下游引物:5’-gtcgacTTAGTGGTGGTGGTGGTGGTGGGTG-3’
(2)加样体系50μL，如下表所示：
1.3.1.2 PCR扩增程序：
1.3.1.3 PCR产物进行胶回收：按照试剂盒说明书操作(可使用天根生化科技有限公司货号为DP214-02的胶回收试剂盒)。
1.3.2 PCR产物及载体双酶切反应
(1)标记好需要用到的200μL PCR管，在PCR管中按照下表进行加样、混匀：50μL反应体系
(2)将步骤(1)中的PCR管置于相应酶最适温度恒温水浴锅中，水浴1-2h。
双酶切产物胶回收：取出上述双酶切体系，进行琼脂糖凝胶电泳以回收其中的DNA片段，方法同1.3.1.3中PCR产物胶回收。
1.3.3连接反应
(1)准备洁净的200μL PCR管若干，做好标记，置于EP管架上待用。
(2)在200μL PCR管按照下表进行加样、混匀。
(3)按照步骤(2)中表格完成加样后，将每个10μl反应体系置于PCR仪16℃反应2h；
(4)取出步骤(3)中的EP管，置于4℃保存。
1.3.4转化反应
将10μL连接反应液快速加入100μL感受态细胞中，并吹打混匀，冰浴30min；取出样品管，置于42℃水浴100s，然后立即冰浴2min；取出样品管，在超净工作台中，向样品管中加入600μL液体LB培养基，然后将样品管置于37℃恒温摇床，220rpm/min，培养1h；涂板：取出上步骤中样品管，室温离心8,000rpm/min，2min，去掉600μL上清液体，剩余上清液重悬管底部的菌体，将重悬的菌液放入相应的转化平板中心，用涂菌棒将转化平板中心的菌液均匀铺开；将上步骤平板正置于生化恒温培养箱中，37℃培养1h后，将转化平板倒置进行培养15h；观察转化结果。转化后阳性质粒命名为pET30a-OPTI-TF-P72。质粒构建示意图如图2所示。
1.3.5质粒抽提与双酶切鉴定
1.3.5.1质粒抽提：按试剂盒照说明书进行操作(可选用生工生物工程(上海)股份有限公司，货号为SK8192的质粒小提试剂盒)。
1.3.5.2双酶切鉴定
(1)标记好需要用到的200μL PCR管，按照下表进行加样：10μL反应体系
(2)将步骤(1)中的200μL PCR管10μL反应体系置于37℃恒温水浴锅中，水浴1h。
(3)将步骤(2)中的酶切体系样品进行琼脂糖凝胶电泳，检查插入片段大小是否正确；实验结果见图3所示，1-4：pET30a-OPTI-TF-P72质粒SacI和SalI双酶切，条带大小分别为6673bp，1979bp，质粒酶切正确。
(4)选择插入片段正确的克隆送测序公司测序。
1.4 P72蛋白表达
1.4.1转化大肠杆菌BL21(DE3)
吸取1μL质粒加入100μL BL21(DE3)感受态细胞中，冰浴30min；42℃热激90s；冰浴2min；在超净台内加入500μL无抗性的LB培养液；37℃220rpm摇1h；吸取100μL菌液涂卡那抗性LB平板，37℃过夜培养，挑取单克隆并加入甘油于-80℃保存备用。
1.4.2小量诱导表达
甘油管保藏管菌株活化：取E.coli BL21(DE3)pET30a-OPTI-TF-P72菌株甘油保藏管解冻，用接种环挑取甘油管中菌悬液于卡那抗性平板(50μg/mL)上划线，37℃培养过夜。挑菌活化：于培养好的平板上挑取单克隆至3mL卡那抗性LB培养基中，37℃220r/min摇床培养3.5～5h，至OD600达到0.5～0.8。发酵接种：将活化好的菌悬液取以1：100的比例接种至15mL卡那抗性LB培养基中，37℃，220r/min培养。降温诱导：当OD 600达到0.6-0.8时，取出5mL菌液，12000rpm离心5min，将菌体置于-20℃保存，即为“诱导前”。剩余菌液置于冰水浴10min，加入0.4ml 1M IPTG，终浓度0.2mM IPTG。将摇床温度降至20℃，诱导15h。菌体收集：发酵结束后，测量OD600，收集与“诱导前”等量菌体，12000r/min离心5min，收集菌体置于-20℃保存，即为“诱导后”。
诱导表达SDS-PAGE检测结果如图4所示，箭头指向为TF-p72蛋白。从图中可以看出，我们制备的TF-P72蛋白为可溶性表达。
另外，参照1.3的步骤，我们也构建了pET28a-OPTI-p72，pQE30Xa-OPTI–p72，pColdIII--OPTI-p72，pBAD30--OPTI–p72等并参照1.4的步骤将该质粒进行了转化和小量诱导表达，发现这些和文献报道基本一致，基本上是不表达。另pET30a-OPTI-MBP-p72、pET30a-OPTI-SUMO-p72，pET30a-OPTI-GST-p72等含有不同伴侣蛋白的质粒，并将该质粒进行了转化和小量诱导表达，发现使用这些质粒制备的蛋白也有不同的表达，后续的实验和生产中用pET30a--OPTI-TF-p72的阳性菌株作为实施例。
1.4.3大量诱导表达
甘油管保藏管菌株活化：取pET30a--OPTI-TF-p72菌株甘油保藏管解冻，用接种环挑取甘油管中菌悬液于卡那抗性平板上划线，37℃培养过夜。挑菌活化：于培养好的平板上挑取单克隆至3mL卡那抗性LB培养基中，37℃220r/min摇床培养5～6h，至OD600达到0.5～0.8。种子液培养：将活化好的菌悬液取450μL接种至450mL卡那抗性LB培养基中，37℃220r/min培养9～10h。发酵培养基配制：按照发酵培养基配方配制发酵培养基组分1于15L发酵罐中，安装组联发酵罐，发酵培养基组分2及补料培养基配制于蓝口瓶中，121℃高压蒸汽灭菌20min。发酵参数设置：Agit 400r/min；Temperature 37℃；pH 7.00；DO 40；Air 100％；Gasflow 2.0。发酵接种：于接种口将450mL发酵培养基组分2，3mL发酵培养基组分3，200μL消泡剂，9mL卡那抗生素(50mg/mL)补加到发酵罐中；将培养好的450mL种子液接种至9L发酵培养基中进行发酵罐放大培养，培养5～6h至OD 600值到12～14。降温诱导：设置温度参数，将发酵罐温度降至20℃，取样，加入2.7mL IPTG(1M)至IPTG终浓度为0.3mmol/L，20℃诱导培养8h。发酵补料：待发酵培养至OD600达到17～19时，按5％的速度连续补加补料培养基(先将补料培养基组分1和2混匀)。菌体收集：发酵结束后，收集发酵液在8000r/min离心10min，收集菌体置于-20℃保存。
其中，上述过程中所用培养基如下：
发酵培养基组分1：酵母粉10g/L，胰蛋白胨20g/L，KH2PO4 1.14g/L，K2HPO4 0.9g/L，(NH4)2SO4 3.0g/L，MgSO4·7H2O 0.3g/L，NaCl 5g/L，pH 7.0；发酵培养基组分2：甘油30g/L；发酵培养基组分3：VB1 2mg/L；补料培养基组分1：酵母粉16.67g/L；胰蛋白胨33.33g/L；补料培养基组分2：甘油100g/L。
1.5 p72蛋白纯化
菌体破碎：向菌体中加入裂解液(10ml/g湿重)，搅拌均匀；将菌体样品倒入到细胞均质仪样品槽中，出样口用烧杯准备收集样品；以样品的90％流出出样口为一个循环，一个循环完成后，将出样口烧杯收集的样品倒回样品槽，共进行4个循环。离心：将上步骤中破碎完全的样品分装到250mL Beckman离心管中，12,000rpm，4℃离心30min，上清作为上样样品。镍柱平衡：用超纯水平衡2～3柱体积(CV)，排出20％的乙醇保存液；然后用裂解液平衡2～3柱体积(CV)。上样：取菌体破碎后上清作为样品与镍柱填料混合，于滚瓶机上混匀1h后进行流穿，并收集流穿液。去除内毒素：用50倍柱体积(CV)的清洗缓冲液组分1洗柱，用于去除内毒素。去除TritonX-114：用30倍柱体积(CV)的洗脱缓冲液组分1(elution buffer 1)(不含Triton X-114)洗柱，减少Triton X-114的残留。洗脱：20mM咪唑洗脱缓冲液洗杂，至考马斯亮蓝G250检测无蓝色；500mM咪唑洗脱缓冲液洗脱目的蛋白，至考马斯亮蓝G250检测无蓝色，收集浓缩目的蛋白。
结果：纯化结果如图5所示，1是Marker，2是纯化后的TF-p72蛋白。从图中可以看出，纯化后的TF-p72蛋白SDS-PAGE纯度均能达到80％以上；经过计算，在没有优化发酵条件的情况下，表达量能达到300mg/L以上。
其中蛋白纯化所需溶液如下：
菌液裂解液：组分为50mM NaH2PO4，500mM NaCl，1mMβ-ME，0.2％TritonX-114，最后调节pH至8.0。清洗缓冲液1：50mM NaH2PO4，1M NaCl，1mMβ-ME，0.2％TritonX-114，最后调节pH至7.0。洗脱缓冲液组分1：50mM NaH2PO4，500mM NaCl，最后调节pH至8.0。洗脱缓冲液组分2：50mM NaH2PO4，500mM NaCl，500mM咪唑，最后调节pH至8.0。
1.6 TF-P72蛋白稳定性
将实施例5纯化后的蛋白用PBS稀释到800ug/ml，共10ml，分成2份，每份5ml；一份置于-80℃冰箱，一份置于4℃冰箱中，每周取样一次，每次0.5ml，连续取样10次；每次取样后用BCA检测蛋白浓度，结果如下表所示：
从蛋白浓度的变化来看，两组实验过程中蛋白基本保持稳定。为了进一步验证处理后的蛋白的含量是否变化，我们用第十次的样品进行SDS-PAGE检测。具体结果如图6所示：1是蛋白Marker 26619，2，3：第十次样品4℃和-20℃蛋白样品，上样量3μg；从图中可以看出，处理后的样品(第十次取样)仍然具有良好的稳定性。
1.7 TF-P72亚单位疫苗的制备
1.7.1疫苗制备
水相准备：根据疫苗中TF-P72蛋白含量，使用PBS(或生理盐水)将TF-P72蛋白稀释适当浓度，即为水相；
油相准备：根据制备疫苗总量，按照抗原相和佐剂重量比为1:1，体积比为46:54，量取适量ISA 201VG佐剂；
乳化：将水相和油相都预热到33℃，将水相缓缓加到油相中，200-500rpm搅拌20-30min，于20℃静置1h置于4℃过夜；
分装、贮存：根据需要进行分装，检合格后于4℃保存备用。
1.7.2疫苗质检
物理性状采用眼观的方法观察外观(是否是乳白色乳剂)；
采用清洁吸管吸取少量疫苗滴于冷水中，观察(除第1滴外)，疫苗应为云雾状扩散，判定为水包油包水剂型；
将疫苗10ml加入离心管中，以3000r/min离心15min，管底析出的水相应≤0.5mL，判为稳定；
使用粘度仪对疫苗进行粘度检测，应在20-50cp，判为合格。
1.7.3 TF-P72亚单位疫苗对小鼠安全性实验
取健康16-20g SPF雌性小鼠(购于浙江中医药大学)20只，分别随机分4个组，每组5只，按照如下方法进行安全性实验。
单剂量一次免疫组：每组5只按肌肉注射接种100μL(25μg/只)，连续观察2周。
单剂量二次免疫组：每组5只按肌肉注射接种100μL(25μg/只)，连续观察2周。2周后以相同方法剂量再接种一次，继续观察2周。
超剂量一次免疫组：每组5只按肌肉注射接种100μL(200μg/只)，连续观察2周。
对照组：每组5只按肌肉注射接种100μL(PBS配制的疫苗)，连续观察2周。
实验期间，每天观察动物的精神，采食、活动、饮水、注射部位炎症变化和排泄情况等临床变化，记录动物的异常情况。
经过连续观察，对注射TF-P72蛋白的小鼠的临床症状进行比较，单剂量，二次免疫剂量，超剂量免疫组和对照组，均饮食正常，精神无不良变化，排泄正常，注射部位未发现发炎现象，无死亡小鼠出现，接种动物没有遇到任何不良反应。说明，本发明制备的疫苗蛋白即使是以高剂量注射免疫(200μg)，也无明显副作用，是一种安全的免疫蛋白。
1.7.4 ELISA检测
(1)包被：用包被液(50mM碳酸盐缓冲液，pH 9.5)将纯化的TF-P72蛋白稀释至0.5μg/ml，每种抗原均包被8孔(4孔加小鼠血清样品，4孔加封闭液作为对照)，每种抗原均加入100μl/孔，封口膜封好后4℃冰箱放置过夜；
(2)洗涤：从冰箱取出酶标板后，用PBST洗板5次；
(3)封闭：每孔加入200μl封闭液(5％脱脂奶)，封口膜封好后37℃孵育2h；
(4)血清稀释：将用TF-P72蛋白免疫后的小鼠的阳性血清(二免14天后的阳性血清)用封闭液稀释500倍(如499μl稀释液中加入1μl血清，混匀即可)；
(5)洗涤：同(2)；
(6)加样：加入稀释血清，同时用封闭液做阴性对照，37℃孵育1h；
(7)洗涤：同(2)；
(8)加二抗：每孔加入稀释(稀释比为1:5000)的HRP标记的兔抗鼠IgG二抗100μl，37℃孵育0.5h；
(9)洗涤：同(2)；
(10)显色：避光条件下每孔加入100μl的TMB显色液，37℃孵育10min；
(11)终止：每孔加入50μl终止液(2M H2SO4)，终止反应；
(12)检测：于450nm波长测定样品OD值，分析数据；
(13)结果如下表所示：包被TF-P72蛋白的能与血清特异性结合，OD450均值为1.116；包被TF-P72蛋白与封闭液均没有特异性结合，OD450均值为0.041。这说明，TF-P72蛋白可以作为Elisa试剂盒的抗原，在摸索合适的包被浓度和血清稀释比后，可以开发一种检测非洲猪瘟感染和免疫的诊断试剂盒。
本发明通过上面的实施例进行举例说明，但是，应当理解，本发明并不限于这里所描述的特殊实例和实施方案。在这里包含这些特殊实例和实施方案的目的在于帮助本领域中的技术人员实践本发明。任何本领域中的技术人员很容易在不脱离本发明精神和范围的情况下进行进一步的改进和完善，因此本发明只受到本发明权利要求的内容和范围的限制，其意图涵盖所有包括在由附录权利要求所限定的本发明精神和范围内的备选方案和等同方案。
序列表
<110> 浙江海隆生物科技有限公司
<120> 一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白及其制备方法和应用
<160> 4
<170> SIPOSequenceListing 1.0
<210> 4
<211> 1941
<212> DNA
<213> 密码子优化后的编码P72蛋白核苷酸序列(DNA)
<400> 4
<210> 4
<211> 1941
<212> DNA
<213> 密码子优化前的编码P72蛋白的核苷酸序列(DNA)
<400> 4
<210> 4
<211> 2046
<212> DNA
<213> 编码TF-P72蛋白的核苷酸序列(DNA)
<400> 4
<210> 4
<211> 1125
<212> PRT
<213> TF-P72蛋白序列(PRT)
<400> 4
Met Asn His Lys Val Met Gln Val Ser Val Glu Thr Thr Gln Gly Leu
1 5 10 15
Gly Arg Arg Val Thr Ile Thr Ile Ala Ala Asp Ser Ile Glu Thr Ala
20 25 30
Val Lys Ser Glu Leu Val Asn Val Ala Lys Lys Val Arg Ile Asp Gly
35 40 45
Phe Arg Lys Gly Lys Val Pro Met Asn Ile Val Ala Gln Arg Tyr Gly
50 55 60
Ala Ser Val Arg Gln Asp Val Leu Gly Asp Leu Met Ser Arg Asn Phe
65 70 75 80
Ile Asp Ala Ile Ile Lys Glu Lys Ile Asn Pro Ala Gly Ala Pro Thr
85 90 95
Tyr Val Pro Gly Glu Tyr Lys Leu Gly Glu Asp Phe Thr Tyr Ser Val
100 105 110
Glu Phe Glu Val Tyr Pro Glu Val Glu Leu Gln Gly Leu Glu Ala Ile
115 120 125
Glu Val Glu Lys Pro Ile Val Glu Val Thr Asp Ala Asp Val Asp Gly
130 135 140
Met Leu Asp Thr Leu Arg Lys Gln Gln Ala Thr Trp Lys Glu Lys Asp
145 150 155 160
Gly Ala Val Glu Ala Glu Asp Arg Val Thr Ile Asp Phe Thr Gly Ser
165 170 175
Val Asp Gly Glu Glu Phe Glu Gly Gly Lys Ala Ser Asp Phe Val Leu
180 185 190
Ala Met Gly Gln Gly Arg Met Ile Pro Gly Phe Glu Asp Gly Ile Lys
195 200 205
Gly His Lys Ala Gly Glu Glu Phe Thr Ile Asp Val Thr Phe Pro Glu
210 215 220
Glu Tyr His Ala Glu Asn Leu Lys Gly Lys Ala Ala Lys Phe Ala Ile
225 230 235 240
Asn Leu Lys Lys Val Glu Glu Arg Glu Leu Pro Glu Leu Thr Ala Glu
245 250 255
Phe Ile Lys Arg Phe Gly Val Glu Asp Gly Ser Val Glu Gly Leu Arg
260 265 270
Ala Glu Val Arg Lys Asn Met Glu Arg Glu Leu Lys Ser Ala Ile Arg
275 280 285
Asn Arg Val Lys Ser Gln Ala Ile Glu Gly Leu Val Lys Ala Asn Asp
290 295 300
Ile Asp Val Pro Ala Ala Leu Ile Asp Ser Glu Ile Asp Val Leu Arg
305 310 315 320
Arg Gln Ala Ala Gln Arg Phe Gly Gly Asn Glu Lys Gln Ala Leu Glu
325 330 335
Leu Pro Arg Glu Leu Phe Glu Glu Gln Ala Lys Arg Arg Val Val Val
340 345 350
Gly Leu Leu Leu Gly Glu Val Ile Arg Thr Asn Glu Leu Lys Ala Asp
355 360 365
Glu Glu Arg Val Lys Gly Leu Ile Glu Glu Met Ala Ser Ala Tyr Glu
370 375 380
Asp Pro Lys Glu Val Ile Glu Phe Tyr Ser Lys Asn Lys Glu Leu Met
385 390 395 400
Asp Asn Met Arg Asn Val Ala Leu Glu Glu Gln Ala Val Glu Ala Val
405 410 415
Leu Ala Lys Ala Lys Val Thr Glu Lys Glu Thr Thr Phe Asn Glu Leu
420 425 430
Met Asn Gln Gln Ala Ser Ala Gly Leu Glu Val Leu Phe Gln Gly Pro
435 440 445
Ser Ala Gly Leu Val Pro Arg Gly Ser Gly Gly Ile Glu Gly Arg His
450 455 460
Met Glu Leu Glu Asn Leu Tyr Phe Gln Gly Ala Ser Gly Gly Ala Phe
465 470 475 480
Cys Leu Ile Ala Asn Asp Gly Lys Ala Asp Lys Ile Ile Leu Ala Gln
485 490 495
Asp Leu Leu Asn Ser Arg Ile Ser Asn Ile Lys Asn Val Asn Lys Ser
500 505 510
Tyr Gly Lys Pro Asp Pro Glu Pro Thr Leu Ser Gln Ile Glu Glu Thr
515 520 525
His Leu Val His Phe Asn Ala His Phe Lys Pro Tyr Val Pro Val Gly
530 535 540
Phe Glu Tyr Asn Lys Val Arg Pro His Thr Gly Thr Pro Thr Leu Gly
545 550 555 560
Asn Lys Leu Thr Phe Gly Ile Pro Gln Tyr Gly Asp Phe Phe His Asp
565 570 575
Met Val Gly His His Ile Leu Gly Ala Cys His Ser Ser Trp Gln Asp
580 585 590
Ala Pro Ile Gln Gly Thr Ser Gln Met Gly Ala His Gly Gln Leu Gln
595 600 605
Thr Phe Pro Arg Asn Gly Tyr Asp Trp Asp Asn Gln Thr Pro Leu Glu
610 615 620
Gly Ala Val Tyr Thr Leu Val Asp Pro Phe Gly Arg Pro Ile Val Pro
625 630 635 640
Gly Thr Lys Asn Ala Tyr Arg Asn Leu Val Tyr Tyr Cys Glu Tyr Pro
645 650 655
Gly Glu Arg Leu Tyr Glu Asn Val Arg Phe Asp Val Asn Gly Asn Ser
660 665 670
Leu Asp Glu Tyr Ser Ser Asp Val Thr Thr Leu Val Arg Lys Phe Cys
675 680 685
Ile Pro Gly Asp Lys Met Thr Gly Tyr Lys His Leu Val Gly Gln Glu
690 695 700
Val Ser Val Glu Gly Thr Ser Gly Pro Leu Leu Cys Asn Ile His Asp
705 710 715 720
Leu His Lys Pro His Gln Ser Lys Pro Ile Leu Thr Asp Glu Asn Asp
725 730 735
Thr Gln Arg Thr Cys Ser His Thr Asn Pro Lys Phe Leu Ser Gln His
740 745 750
Phe Pro Glu Asn Ser His Asn Ile Gln Thr Ala Gly Lys Gln Asp Ile
755 760 765
Thr Pro Ile Thr Asp Ala Thr Tyr Leu Asp Ile Arg Arg Asn Val His
770 775 780
Tyr Ser Cys Asn Gly Pro Gln Thr Pro Lys Tyr Tyr Gln Pro Pro Leu
785 790 795 800
Ala Leu Trp Ile Lys Leu Arg Phe Trp Phe Asn Glu Asn Val Asn Leu
805 810 815
Ala Ile Pro Ser Val Ser Ile Pro Phe Gly Glu Arg Phe Ile Thr Ile
820 825 830
Lys Leu Ala Ser Gln Lys Asp Leu Val Asn Glu Phe Pro Gly Leu Phe
835 840 845
Val Arg Gln Ser Arg Phe Ile Ala Gly Arg Pro Ser Arg Arg Asn Ile
850 855 860
Arg Phe Lys Pro Trp Phe Ile Pro Gly Val Ile Asn Glu Ile Ser Leu
865 870 875 880
Thr Asn Asn Glu Leu Tyr Ile Asn Asn Leu Phe Val Thr Pro Glu Ile
885 890 895
His Asn Leu Phe Val Lys Arg Val Arg Phe Ser Leu Ile Arg Val His
900 905 910
Lys Thr Gln Val Thr His Thr Asn Asn Asn His His Asp Glu Lys Leu
915 920 925
Met Ser Ala Leu Lys Trp Pro Ile Glu Tyr Met Phe Ile Gly Leu Lys
930 935 940
Pro Thr Trp Asn Ile Ser Asp Gln Asn Pro His Gln His Arg Asp Trp
945 950 955 960
His Lys Phe Gly His Val Val Asn Ala Ile Met Gln Pro Thr His His
965 970 975
Ala Glu Ile Ser Phe Gln Asp Arg Asp Thr Ala Leu Pro Asp Ala Cys
980 985 990
Ser Ser Ile Ser Asp Ile Ser Pro Val Thr Tyr Pro Ile Thr Leu Pro
995 1000 1005
Ile Ile Lys Asn Ile Ser Val Thr Ala His Gly Ile Asn Leu Ile Asp
1010 1015 1020
Lys Phe Pro Ser Lys Phe Cys Ser Ser Tyr Ile Pro Phe His Tyr Gly
1025 1030 1035 1040
Gly Asn Ala Ile Lys Thr Pro Asp Asp Pro Gly Ala Met Met Ile Thr
1045 1050 1055
Phe Ala Leu Lys Pro Arg Glu Glu Tyr Gln Pro Ser Gly His Ile Asn
1060 1065 1070
Val Ser Arg Ala Arg Glu Phe Tyr Ile Ser Trp Asp Thr Asp Tyr Val
1075 1080 1085
Gly Ser Ile Thr Thr Ala Asp Leu Val Val Ser Ala Ser Ala Ile Asn
1090 1095 1100
Phe Leu Leu Leu Gln Asn Gly Ser Ala Val Leu Arg Tyr Ser Thr
1105 1110 1115
Claims (11)
1.一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白，其特征在于，所述非洲猪瘟病毒p72亚单位可溶性融合蛋白包括OPTI-p72蛋白和一种伴侣蛋白，其中所述OPTI-p72蛋白的核苷酸序列为p72蛋白经优化后的序列OPTI-p72如SEQ ID NO.1所示，所述伴侣蛋白为TF蛋白。
2.根据权利要求1所述的非洲猪瘟病毒p72亚单位可溶性融合蛋白，其特征在于，所述非洲猪瘟病毒p72亚单位可溶性融合蛋白的核苷酸序列OPTI-TF-p72是OPTI-p72蛋白的5，端插入TF蛋白的核苷酸序列，所述OPTI-TF-p72蛋白的核苷酸序列如SEQ ID NO.3所示。
3.根据权利要求2所述的非洲猪瘟病毒p72亚单位可溶性融合蛋白，其特征在于，所述非洲猪瘟病毒p72亚单位可溶性融合蛋白的氨基酸序列如SEQ ID NO.4所示。
4.根据权利要求3所述的非洲猪瘟病毒p72亚单位可溶性融合蛋白，其特征在于，所述非洲猪瘟病毒p72亚单位可溶性融合蛋白在如SEQ ID NO.4所示的氨基酸序列的氨基末端或羧基末端连接有poly-His、FLAG、c-myc、HA和poly-Arg中的一种标签。
5.一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白的制备方法，其特征在于，所述制备方法包括以下步骤：
1)将含有OPTI-p72蛋白和一种伴侣蛋白的基因克隆到原核表达载体中，得到含有非洲猪瘟病毒OPTI-p72蛋白和一种伴侣蛋白编码基因的重组质粒；其中，所述OPTI-p72蛋白的核苷酸序列为p72蛋白经优化后的序列OPTI-p72如SEQ ID NO.1所示，所述伴侣蛋白为TF蛋白；
2)再将步骤1)所述重组质粒转化至大肠杆菌菌株的细胞中，得到重组表达的菌株；
3)通过培养、筛选、步骤2)中所述重组表达的菌株，得到高度表达的菌株；
4)发酵培养步骤3)中所述高度表达的菌株，纯化后得到重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白。
6.根据权利要求5所述的制备方法，其特征在于，含有非洲猪瘟病毒OPTI-p72蛋白和TF蛋白的核苷酸序列OPTI-TF-p72是OPTI-p72蛋白的5，端插入TF蛋白的核苷酸序列，所述OPTI-TF-p72蛋白的核苷酸序列如SEQ ID NO.3所示。
7.根据权利要求6所述的制备方法，其特征在于，其特征在于，含有非洲猪瘟病毒OPTI-p72蛋白和TF蛋白的的氨基酸序列如SEQ ID NO.4所示。
8.根据权利要求7所述的制备方法，其特征在于，其特征在于，含有非洲猪瘟病毒OPTI-p72蛋白和TF蛋白在如SEQ ID NO.4所示的氨基酸序列的氨基末端或羧基末端连接有poly-His、FLAG、c-myc、HA和poly-Arg中的一种标签。
9.根据权利要求5所述的制备方法，其特征在于，步骤1)中，所述原核表达载体为pET28a、pET30a、pBAD、pcold、pQE、或pKK。
10.根据权利要求5所述的制备方法，其特征在于，步骤2)中，所述大肠杆菌菌株选自BL21(DE3)、BL21 star(DE3)、arcticexpress中的一种菌株。
11.一种如权利要求1～4任一所述的非洲猪瘟病毒p72亚单位可溶性融合蛋白在制备诊断、预防和治疗非洲猪瘟的疫苗中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910142648.5A CN111607001B (zh) | 2019-02-26 | 2019-02-26 | 一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910142648.5A CN111607001B (zh) | 2019-02-26 | 2019-02-26 | 一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白及其制备方法和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111607001A CN111607001A (zh) | 2020-09-01 |
| CN111607001B true CN111607001B (zh) | 2023-04-25 |
Family
ID=72197564
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910142648.5A Active CN111607001B (zh) | 2019-02-26 | 2019-02-26 | 一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111607001B (zh) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112274502A (zh) * | 2020-10-28 | 2021-01-29 | 河南佰景生物科技有限公司 | 一种氨基酸衍生物在制备防治非洲猪瘟的制剂中的应用 |
| CN117286185A (zh) * | 2021-05-13 | 2023-12-26 | 浙江海隆生物科技有限公司 | 一种重组的非洲猪瘟病毒p72三聚体亚单位蛋白的制备方法及其应用 |
| CN112979765B (zh) * | 2021-05-13 | 2021-08-03 | 中国农业大学 | 一种非洲猪瘟病毒衣壳蛋白p72及其制备方法和应用 |
| CN113322268A (zh) * | 2021-05-26 | 2021-08-31 | 中国农业大学 | 一种非洲猪瘟病毒p72重组蛋白及其构建的胶体金免疫层析试纸 |
| CN113755513A (zh) * | 2021-09-07 | 2021-12-07 | 天津大学 | 一种基于非洲猪瘟病毒p30、p72蛋白的病毒样颗粒的制备与应用 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754981A (zh) * | 2017-01-23 | 2017-05-31 | 山东省滨州畜牧兽医研究院 | 一种利用大肠杆菌系统制备小鹅瘟病毒样颗粒的方法 |
| 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 |
|---|---|---|---|---|
| ES2337113B1 (es) * | 2007-04-17 | 2011-01-24 | Centre De Recerca En Sanitat Animal (Cresa) | Empleo de la hemaglutinina del virus de la peste porcina africana como adyuvante. |
-
2019
- 2019-02-26 CN CN201910142648.5A patent/CN111607001B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754981A (zh) * | 2017-01-23 | 2017-05-31 | 山东省滨州畜牧兽医研究院 | 一种利用大肠杆菌系统制备小鹅瘟病毒样颗粒的方法 |
| 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 |
Non-Patent Citations (3)
| Title |
|---|
| African swine fever virus Georgia 2007/1 complete genome;FR682468;《Genebank Database》;20160725;第1-75页 * |
| Purification of inclusion body−forming peptides and proteins in soluble form by fusion to Escherichia coli thermostable proteins;Arjun Thapa等;《BioTechniques》;20080531;第787-796页 * |
| The African Swine Fever Virus Nonstructural Protein pB602L Is Required for Formation of the Icosahedral Capsid of the Virus Particle;Carolina Epifano等;《JOURNAL OF VIROLOGY》;20061231;第12260-12270页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111607001A (zh) | 2020-09-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111607001B (zh) | 一种重组的非洲猪瘟病毒p72亚单位可溶性融合蛋白及其制备方法和应用 | |
| CN111393531B (zh) | 一种亚单位融合蛋白CD2V-Fc及其制备方法和应用 | |
| CN111777672A (zh) | 一种重组的非洲猪瘟病毒pKP177R亚单位可溶性蛋白及其制备方法和应用 | |
| CN107987135B (zh) | Ι群4型禽腺病毒亚单位蛋白、其制备方法及应用 | |
| CN110041411B (zh) | 稳定的非典型猪瘟病毒亚单位蛋白及其疫苗和制备方法和应用 | |
| CN112979765B (zh) | 一种非洲猪瘟病毒衣壳蛋白p72及其制备方法和应用 | |
| CN116217737A (zh) | 非洲猪瘟病毒免疫原的p49突变融合蛋白、重组载体、工程菌及制备方法和应用 | |
| CN110183520B (zh) | 一种猪丹毒SpaA蛋白及其在制备疫苗中的应用 | |
| CN111808176B (zh) | 牛疱疹病毒抗原组合物及其应用 | |
| CN111607000B (zh) | 一种重组的非洲猪瘟病毒p30亚单位可溶性融合蛋白及其制备方法和应用 | |
| CN114874995B (zh) | 猪瘟病毒2型Erns蛋白的单克隆抗体杂交瘤细胞株及应用 | |
| CN114163505B (zh) | 猪瘟、猪伪狂犬病毒二联亚单位疫苗及其制备方法 | |
| CN112142851B (zh) | 一种狂犬病毒表面的亚单位融合蛋白tG及其制备方法和应用 | |
| CN116478953B (zh) | 鲍曼不动杆菌DlaT重组蛋白、制备方法及应用 | |
| CN111440815B (zh) | 一种新型鸭呼肠孤病毒的复合疫苗及卵黄抗体制备方法 | |
| CN108578686A (zh) | 一种制备羊梭菌病基因工程亚单位疫苗的方法 | |
| CN110746496B (zh) | 一种鲍曼不动杆菌的pal重组蛋白及其编码基因和它们的应用 | |
| CN112341525B (zh) | 一种重组非洲猪瘟病毒pE120R亚单位可溶性蛋白及其制备方法和应用 | |
| CN114437235B (zh) | 马链球菌马亚种8种蛋白的重组融合蛋白及其制备方法和应用 | |
| CN110845624A (zh) | 一种sumo-cp融合蛋白及其制备方法以及其多克隆抗体的制备方法 | |
| CN115073559A (zh) | 重组的非洲猪瘟病毒ep153r亚单位跨膜蛋白的原核可溶性表达方法和应用 | |
| CN113735943A (zh) | 一种重组的非洲猪瘟病毒p72亚单位蛋白和制备方法及其应用 | |
| CN113817027A (zh) | 一种牛疱疹病毒Ⅰ型gE蛋白胞外区的原核可溶性表达方法 | |
| CN113861277A (zh) | 一种牛轮状病毒重组vp8蛋白与应用 | |
| CN109535229B (zh) | 重组猪圆环病毒2型Rep蛋白、ELISA检测试剂盒及其制备方法与应用 |
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 | ||
| CB02 | Change of applicant information |
Address after: 312366 No. 1, Baichuan Road, Binhai New Area, Shaoxing City, Zhejiang Province Applicant after: NOVO BIOTECH Corp. Address before: 312000 5th floor, building 2, science and innovation center, 398 mahuan Road, Binhai New Town, Shaoxing City, Zhejiang Province Applicant before: NOVO BIOTECH Corp. |
|
| CB02 | Change of applicant information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP03 | Change of name, title or address |
Address after: 312366 No. 1, Baichuan Road, Binhai New Area, Shaoxing City, Zhejiang Province Patentee after: Zhejiang Hailong Biotechnology Co.,Ltd. Country or region after: China Address before: 312366 No. 1, Baichuan Road, Binhai New Area, Shaoxing City, Zhejiang Province Patentee before: NOVO BIOTECH Corp. Country or region before: China |