CN113501885B - 嵌合抗病基因及其相关生物材料与应用 - Google Patents
嵌合抗病基因及其相关生物材料与应用 Download PDFInfo
- Publication number
- CN113501885B CN113501885B CN202110834140.9A CN202110834140A CN113501885B CN 113501885 B CN113501885 B CN 113501885B CN 202110834140 A CN202110834140 A CN 202110834140A CN 113501885 B CN113501885 B CN 113501885B
- Authority
- CN
- China
- Prior art keywords
- tlr2
- leu
- protein
- seq
- gene
- 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
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 92
- 239000012620 biological material Substances 0.000 title claims abstract description 16
- 201000010099 disease Diseases 0.000 title abstract description 13
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title abstract description 13
- 210000004027 cell Anatomy 0.000 claims abstract description 59
- 239000013598 vector Substances 0.000 claims abstract description 56
- 241000283707 Capra Species 0.000 claims abstract description 41
- 210000002540 macrophage Anatomy 0.000 claims abstract description 41
- 108020000411 Toll-like receptor Proteins 0.000 claims abstract description 35
- 102000002689 Toll-like receptor Human genes 0.000 claims abstract description 34
- 102000037865 fusion proteins Human genes 0.000 claims abstract description 32
- 108020001507 fusion proteins Proteins 0.000 claims abstract description 32
- 101000831567 Homo sapiens Toll-like receptor 2 Proteins 0.000 claims abstract description 31
- 102100024333 Toll-like receptor 2 Human genes 0.000 claims abstract description 30
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 28
- 230000003834 intracellular effect Effects 0.000 claims abstract description 18
- 206010057249 Phagocytosis Diseases 0.000 claims abstract description 11
- 230000008782 phagocytosis Effects 0.000 claims abstract description 11
- 230000036039 immunity Effects 0.000 claims abstract description 5
- 208000035240 Disease Resistance Diseases 0.000 claims abstract description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 5
- 102000004169 proteins and genes Human genes 0.000 claims description 49
- 108020004414 DNA Proteins 0.000 claims description 16
- 230000014509 gene expression Effects 0.000 claims description 16
- 102000039446 nucleic acids Human genes 0.000 claims description 15
- 108020004707 nucleic acids Proteins 0.000 claims description 15
- 150000007523 nucleic acids Chemical class 0.000 claims description 15
- 241001465754 Metazoa Species 0.000 claims description 14
- 239000002773 nucleotide Substances 0.000 claims description 13
- 125000003729 nucleotide group Chemical group 0.000 claims description 13
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 11
- 101000669447 Homo sapiens Toll-like receptor 4 Proteins 0.000 claims description 10
- 102000053602 DNA Human genes 0.000 claims description 9
- 102100039360 Toll-like receptor 4 Human genes 0.000 claims description 9
- 244000005700 microbiome Species 0.000 claims description 7
- 230000002708 enhancing effect Effects 0.000 claims description 5
- 108091026890 Coding region Proteins 0.000 claims description 4
- 210000004102 animal cell Anatomy 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims 1
- 210000002950 fibroblast Anatomy 0.000 abstract description 18
- 102000004127 Cytokines Human genes 0.000 abstract description 7
- 108090000695 Cytokines Proteins 0.000 abstract description 7
- 102000007863 pattern recognition receptors Human genes 0.000 abstract description 7
- 108010089193 pattern recognition receptors Proteins 0.000 abstract description 7
- 244000052616 bacterial pathogen Species 0.000 abstract description 6
- 208000004396 mastitis Diseases 0.000 abstract description 3
- 210000001539 phagocyte Anatomy 0.000 abstract description 3
- 239000005482 chemotactic factor Substances 0.000 abstract description 2
- 150000001413 amino acids Chemical group 0.000 description 19
- 230000009261 transgenic effect Effects 0.000 description 15
- 108091034057 RNA (poly(A)) Proteins 0.000 description 13
- 239000013612 plasmid Substances 0.000 description 12
- 238000000034 method Methods 0.000 description 10
- 108091028043 Nucleic acid sequence Proteins 0.000 description 9
- 230000006870 function Effects 0.000 description 9
- 108091033409 CRISPR Proteins 0.000 description 8
- 238000010367 cloning Methods 0.000 description 8
- 230000010354 integration Effects 0.000 description 8
- 230000008685 targeting Effects 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- 101150076937 TLR2 gene Proteins 0.000 description 7
- 239000003446 ligand Substances 0.000 description 7
- 238000011529 RT qPCR Methods 0.000 description 6
- 238000010362 genome editing Methods 0.000 description 6
- 108091008146 restriction endonucleases Proteins 0.000 description 6
- 238000011144 upstream manufacturing Methods 0.000 description 6
- OEDPHAKKZGDBEV-GFPBKZJXSA-N (2s)-6-amino-2-[[(2s)-6-amino-2-[[(2s)-6-amino-2-[[(2s)-6-amino-2-[[(2s)-2-[[(2r)-3-[2,3-di(hexadecanoyloxy)propylsulfanyl]-2-(hexadecanoylamino)propanoyl]amino]-3-hydroxypropanoyl]amino]hexanoyl]amino]hexanoyl]amino]hexanoyl]amino]hexanoic acid Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)CCCCCCCCCCCCCCC)CSCC(COC(=O)CCCCCCCCCCCCCCC)OC(=O)CCCCCCCCCCCCCCC OEDPHAKKZGDBEV-GFPBKZJXSA-N 0.000 description 5
- 108010084333 N-palmitoyl-S-(2,3-bis(palmitoyloxy)propyl)cysteinyl-seryl-lysyl-lysyl-lysyl-lysine Proteins 0.000 description 5
- 101800000795 Proadrenomedullin N-20 terminal peptide Proteins 0.000 description 5
- 102400001018 Proadrenomedullin N-20 terminal peptide Human genes 0.000 description 5
- 108091027544 Subgenomic mRNA Proteins 0.000 description 5
- 108010034529 leucyl-lysine Proteins 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 108090001005 Interleukin-6 Proteins 0.000 description 4
- 102000003960 Ligases Human genes 0.000 description 4
- 108090000364 Ligases Proteins 0.000 description 4
- 125000000539 amino acid group Chemical group 0.000 description 4
- 238000000137 annealing Methods 0.000 description 4
- 238000010276 construction Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 239000002158 endotoxin Substances 0.000 description 4
- 238000001976 enzyme digestion Methods 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 210000005007 innate immune system Anatomy 0.000 description 4
- 229920006008 lipopolysaccharide Polymers 0.000 description 4
- 230000001404 mediated effect Effects 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 230000019491 signal transduction Effects 0.000 description 4
- 238000001890 transfection Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- UMHUHHJMEXNSIV-CIUDSAMLSA-N Asp-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UMHUHHJMEXNSIV-CIUDSAMLSA-N 0.000 description 3
- 238000010354 CRISPR gene editing Methods 0.000 description 3
- 108010012236 Chemokines Proteins 0.000 description 3
- 102000019034 Chemokines Human genes 0.000 description 3
- 108090001007 Interleukin-8 Proteins 0.000 description 3
- IOQWIOPSKJOEKI-SRVKXCTJSA-N Lys-Ser-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O IOQWIOPSKJOEKI-SRVKXCTJSA-N 0.000 description 3
- 241001494479 Pecora Species 0.000 description 3
- 101150063416 add gene Proteins 0.000 description 3
- 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 3
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 3
- 230000030279 gene silencing Effects 0.000 description 3
- 238000010363 gene targeting Methods 0.000 description 3
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 3
- 230000009545 invasion Effects 0.000 description 3
- 108010057821 leucylproline Proteins 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 108020003175 receptors Proteins 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 241000588986 Alcaligenes Species 0.000 description 2
- 208000031295 Animal disease Diseases 0.000 description 2
- DAPLJWATMAXPPZ-CIUDSAMLSA-N Asn-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(N)=O DAPLJWATMAXPPZ-CIUDSAMLSA-N 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 241000588698 Erwinia Species 0.000 description 2
- 241000588722 Escherichia Species 0.000 description 2
- 108700024394 Exon Proteins 0.000 description 2
- 241000589565 Flavobacterium Species 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101000763579 Homo sapiens Toll-like receptor 1 Proteins 0.000 description 2
- 101000669406 Homo sapiens Toll-like receptor 6 Proteins 0.000 description 2
- 102100026720 Interferon beta Human genes 0.000 description 2
- 108090000467 Interferon-beta Proteins 0.000 description 2
- 108050006617 Interleukin-1 receptor Proteins 0.000 description 2
- 102000019223 Interleukin-1 receptor Human genes 0.000 description 2
- QWWPYKKLXWOITQ-VOAKCMCISA-N Leu-Thr-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(C)C QWWPYKKLXWOITQ-VOAKCMCISA-N 0.000 description 2
- WVJNGSFKBKOKRV-AJNGGQMLSA-N Lys-Leu-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WVJNGSFKBKOKRV-AJNGGQMLSA-N 0.000 description 2
- 101710175625 Maltose/maltodextrin-binding periplasmic protein Proteins 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- YCCUXNNKXDGMAM-KKUMJFAQSA-N Phe-Leu-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YCCUXNNKXDGMAM-KKUMJFAQSA-N 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- KNCJWSPMTFFJII-ZLUOBGJFSA-N Ser-Cys-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(O)=O)C(O)=O KNCJWSPMTFFJII-ZLUOBGJFSA-N 0.000 description 2
- 238000000692 Student's t-test Methods 0.000 description 2
- 101150082427 Tlr4 gene Proteins 0.000 description 2
- 102100027010 Toll-like receptor 1 Human genes 0.000 description 2
- 102100039387 Toll-like receptor 6 Human genes 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000000941 anti-staphylcoccal effect Effects 0.000 description 2
- 108010047857 aspartylglycine Proteins 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 108010045262 enhanced cyan fluorescent protein Proteins 0.000 description 2
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 108010012058 leucyltyrosine Proteins 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- DAZSWUUAFHBCGE-KRWDZBQOSA-N n-[(2s)-3-methyl-1-oxo-1-pyrrolidin-1-ylbutan-2-yl]-3-phenylpropanamide Chemical compound N([C@@H](C(C)C)C(=O)N1CCCC1)C(=O)CCC1=CC=CC=C1 DAZSWUUAFHBCGE-KRWDZBQOSA-N 0.000 description 2
- 238000010449 nuclear transplantation Methods 0.000 description 2
- 230000000242 pagocytic effect Effects 0.000 description 2
- 108010051242 phenylalanylserine Proteins 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 108010061238 threonyl-glycine Proteins 0.000 description 2
- 108010084932 tryptophyl-proline Proteins 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 241000589158 Agrobacterium Species 0.000 description 1
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 1
- STACJSVFHSEZJV-GHCJXIJMSA-N Ala-Asn-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O STACJSVFHSEZJV-GHCJXIJMSA-N 0.000 description 1
- WCBVQNZTOKJWJS-ACZMJKKPSA-N Ala-Cys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(O)=O WCBVQNZTOKJWJS-ACZMJKKPSA-N 0.000 description 1
- CSAHOYQKNHGDHX-ACZMJKKPSA-N Ala-Gln-Asn Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CSAHOYQKNHGDHX-ACZMJKKPSA-N 0.000 description 1
- XYTNPQNAZREREP-XQXXSGGOSA-N Ala-Glu-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XYTNPQNAZREREP-XQXXSGGOSA-N 0.000 description 1
- PNALXAODQKTNLV-JBDRJPRFSA-N Ala-Ile-Ala Chemical compound C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O PNALXAODQKTNLV-JBDRJPRFSA-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
- IHMCQESUJVZTKW-UBHSHLNASA-N Ala-Phe-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](C)N)CC1=CC=CC=C1 IHMCQESUJVZTKW-UBHSHLNASA-N 0.000 description 1
- KUFVXLQLDHJVOG-SHGPDSBTSA-N Ala-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](C)N)O KUFVXLQLDHJVOG-SHGPDSBTSA-N 0.000 description 1
- IIABBYGHLYWVOS-FXQIFTODSA-N Arg-Asn-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O IIABBYGHLYWVOS-FXQIFTODSA-N 0.000 description 1
- ZTKHZAXGTFXUDD-VEVYYDQMSA-N Arg-Asn-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZTKHZAXGTFXUDD-VEVYYDQMSA-N 0.000 description 1
- RWDVGVPHEWOZMO-GUBZILKMSA-N Arg-Cys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CS)NC(=O)[C@@H](N)CCCNC(N)=N)C(O)=O RWDVGVPHEWOZMO-GUBZILKMSA-N 0.000 description 1
- COXMUHNBYCVVRG-DCAQKATOSA-N Arg-Leu-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O COXMUHNBYCVVRG-DCAQKATOSA-N 0.000 description 1
- CGWVCWFQGXOUSJ-ULQDDVLXSA-N Arg-Tyr-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(O)=O CGWVCWFQGXOUSJ-ULQDDVLXSA-N 0.000 description 1
- MEFGKQUUYZOLHM-GMOBBJLQSA-N Asn-Arg-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MEFGKQUUYZOLHM-GMOBBJLQSA-N 0.000 description 1
- MFFOYNGMOYFPBD-DCAQKATOSA-N Asn-Arg-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O MFFOYNGMOYFPBD-DCAQKATOSA-N 0.000 description 1
- NVGWESORMHFISY-SRVKXCTJSA-N Asn-Asn-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O NVGWESORMHFISY-SRVKXCTJSA-N 0.000 description 1
- YYSYDIYQTUPNQQ-SXTJYALSSA-N Asn-Ile-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YYSYDIYQTUPNQQ-SXTJYALSSA-N 0.000 description 1
- 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 1
- HDHZCEDPLTVHFZ-GUBZILKMSA-N Asn-Leu-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O HDHZCEDPLTVHFZ-GUBZILKMSA-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
- DJIMLSXHXKWADV-CIUDSAMLSA-N Asn-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(N)=O DJIMLSXHXKWADV-CIUDSAMLSA-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
- XMHFCUKJRCQXGI-CIUDSAMLSA-N Asn-Pro-Gln Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC(=O)N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O XMHFCUKJRCQXGI-CIUDSAMLSA-N 0.000 description 1
- VLDRQOHCMKCXLY-SRVKXCTJSA-N Asn-Ser-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O VLDRQOHCMKCXLY-SRVKXCTJSA-N 0.000 description 1
- PIABYSIYPGLLDQ-XVSYOHENSA-N Asn-Thr-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PIABYSIYPGLLDQ-XVSYOHENSA-N 0.000 description 1
- ZELQAFZSJOBEQS-ACZMJKKPSA-N Asp-Asn-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ZELQAFZSJOBEQS-ACZMJKKPSA-N 0.000 description 1
- AKPLMZMNJGNUKT-ZLUOBGJFSA-N Asp-Asp-Cys Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CS)C(O)=O AKPLMZMNJGNUKT-ZLUOBGJFSA-N 0.000 description 1
- NYQHSUGFEWDWPD-ACZMJKKPSA-N Asp-Gln-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC(=O)O)N NYQHSUGFEWDWPD-ACZMJKKPSA-N 0.000 description 1
- KLYPOCBLKMPBIQ-GHCJXIJMSA-N Asp-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC(=O)O)N KLYPOCBLKMPBIQ-GHCJXIJMSA-N 0.000 description 1
- JNNVNVRBYUJYGS-CIUDSAMLSA-N Asp-Leu-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O JNNVNVRBYUJYGS-CIUDSAMLSA-N 0.000 description 1
- XLILXFRAKOYEJX-GUBZILKMSA-N Asp-Leu-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O XLILXFRAKOYEJX-GUBZILKMSA-N 0.000 description 1
- PCJOFZYFFMBZKC-PCBIJLKTSA-N Asp-Phe-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O PCJOFZYFFMBZKC-PCBIJLKTSA-N 0.000 description 1
- UTLCRGFJFSZWAW-OLHMAJIHSA-N Asp-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)O)N)O UTLCRGFJFSZWAW-OLHMAJIHSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 238000010453 CRISPR/Cas method Methods 0.000 description 1
- 241000283708 Capra aegagrus Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- AVFGSUXQKHIQJS-QEJZJMRPSA-N Cys-Gln-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CS)N)C(O)=O)=CNC2=C1 AVFGSUXQKHIQJS-QEJZJMRPSA-N 0.000 description 1
- KGIHMGPYGXBYJJ-SRVKXCTJSA-N Cys-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CS KGIHMGPYGXBYJJ-SRVKXCTJSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 102100031780 Endonuclease Human genes 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- LJEPDHWNQXPXMM-NHCYSSNCSA-N Gln-Arg-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(O)=O LJEPDHWNQXPXMM-NHCYSSNCSA-N 0.000 description 1
- CYTSBCIIEHUPDU-ACZMJKKPSA-N Gln-Asp-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O CYTSBCIIEHUPDU-ACZMJKKPSA-N 0.000 description 1
- UFNSPPFJOHNXRE-AUTRQRHGSA-N Gln-Gln-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O UFNSPPFJOHNXRE-AUTRQRHGSA-N 0.000 description 1
- KCJJFESQRXGTGC-BQBZGAKWSA-N Gln-Glu-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O KCJJFESQRXGTGC-BQBZGAKWSA-N 0.000 description 1
- HHQCBFGKQDMWSP-GUBZILKMSA-N Gln-Leu-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCC(=O)N)N HHQCBFGKQDMWSP-GUBZILKMSA-N 0.000 description 1
- FKXCBKCOSVIGCT-AVGNSLFASA-N Gln-Lys-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O FKXCBKCOSVIGCT-AVGNSLFASA-N 0.000 description 1
- NYCVMJGIJYQWDO-CIUDSAMLSA-N Gln-Ser-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NYCVMJGIJYQWDO-CIUDSAMLSA-N 0.000 description 1
- RSUVOPBMWMTVDI-XEGUGMAKSA-N Glu-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCC(O)=O)C)C(O)=O)=CNC2=C1 RSUVOPBMWMTVDI-XEGUGMAKSA-N 0.000 description 1
- JRCUFCXYZLPSDZ-ACZMJKKPSA-N Glu-Asp-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O JRCUFCXYZLPSDZ-ACZMJKKPSA-N 0.000 description 1
- HILMIYALTUQTRC-XVKPBYJWSA-N Glu-Gly-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O HILMIYALTUQTRC-XVKPBYJWSA-N 0.000 description 1
- ZWABFSSWTSAMQN-KBIXCLLPSA-N Glu-Ile-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O ZWABFSSWTSAMQN-KBIXCLLPSA-N 0.000 description 1
- BKRQSECBKKCCKW-HVTMNAMFSA-N Glu-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](CCC(=O)O)N BKRQSECBKKCCKW-HVTMNAMFSA-N 0.000 description 1
- IRXNJYPKBVERCW-DCAQKATOSA-N Glu-Leu-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IRXNJYPKBVERCW-DCAQKATOSA-N 0.000 description 1
- MWMJCGBSIORNCD-AVGNSLFASA-N Glu-Leu-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O MWMJCGBSIORNCD-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
- CUPSDFQZTVVTSK-GUBZILKMSA-N Glu-Lys-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCC(O)=O CUPSDFQZTVVTSK-GUBZILKMSA-N 0.000 description 1
- RBXSZQRSEGYDFG-GUBZILKMSA-N Glu-Lys-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O RBXSZQRSEGYDFG-GUBZILKMSA-N 0.000 description 1
- QDMVXRNLOPTPIE-WDCWCFNPSA-N Glu-Lys-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QDMVXRNLOPTPIE-WDCWCFNPSA-N 0.000 description 1
- JWNZHMSRZXXGTM-XKBZYTNZSA-N Glu-Ser-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JWNZHMSRZXXGTM-XKBZYTNZSA-N 0.000 description 1
- YQAQQKPWFOBSMU-WDCWCFNPSA-N Glu-Thr-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O YQAQQKPWFOBSMU-WDCWCFNPSA-N 0.000 description 1
- MXJYXYDREQWUMS-XKBZYTNZSA-N Glu-Thr-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O MXJYXYDREQWUMS-XKBZYTNZSA-N 0.000 description 1
- UUTGYDAKPISJAO-JYJNAYRXSA-N Glu-Tyr-Leu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C(O)=O)CC1=CC=C(O)C=C1 UUTGYDAKPISJAO-JYJNAYRXSA-N 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- XCLCVBYNGXEVDU-WHFBIAKZSA-N Gly-Asn-Ser Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O XCLCVBYNGXEVDU-WHFBIAKZSA-N 0.000 description 1
- GRIRDMVMJJDZKV-RCOVLWMOSA-N Gly-Asn-Val Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O GRIRDMVMJJDZKV-RCOVLWMOSA-N 0.000 description 1
- XLFHCWHXKSFVIB-BQBZGAKWSA-N Gly-Gln-Gln Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O XLFHCWHXKSFVIB-BQBZGAKWSA-N 0.000 description 1
- HFXJIZNEXNIZIJ-BQBZGAKWSA-N Gly-Glu-Gln Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O HFXJIZNEXNIZIJ-BQBZGAKWSA-N 0.000 description 1
- KMSGYZQRXPUKGI-BYPYZUCNSA-N Gly-Gly-Asn Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC(N)=O KMSGYZQRXPUKGI-BYPYZUCNSA-N 0.000 description 1
- HMHRTKOWRUPPNU-RCOVLWMOSA-N Gly-Ile-Gly Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O HMHRTKOWRUPPNU-RCOVLWMOSA-N 0.000 description 1
- AAHSHTLISQUZJL-QSFUFRPTSA-N Gly-Ile-Ile Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O AAHSHTLISQUZJL-QSFUFRPTSA-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
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- HVLSXIKZNLPZJJ-TXZCQADKSA-N HA peptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HVLSXIKZNLPZJJ-TXZCQADKSA-N 0.000 description 1
- GMIWMPUGTFQFHK-KCTSRDHCSA-N His-Ala-Trp Chemical compound C[C@H](NC(=O)[C@@H](N)Cc1cnc[nH]1)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(O)=O GMIWMPUGTFQFHK-KCTSRDHCSA-N 0.000 description 1
- JBJNKUOMNZGQIM-PYJNHQTQSA-N His-Arg-Ile Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JBJNKUOMNZGQIM-PYJNHQTQSA-N 0.000 description 1
- 108010093488 His-His-His-His-His-His Proteins 0.000 description 1
- BXOLYFJYQQRQDJ-MXAVVETBSA-N His-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC1=CN=CN1)N BXOLYFJYQQRQDJ-MXAVVETBSA-N 0.000 description 1
- SKOKHBGDXGTDDP-MELADBBJSA-N His-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N SKOKHBGDXGTDDP-MELADBBJSA-N 0.000 description 1
- YXXKBPJEIYFGOD-MGHWNKPDSA-N His-Phe-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CC2=CN=CN2)N YXXKBPJEIYFGOD-MGHWNKPDSA-N 0.000 description 1
- FHKZHRMERJUXRJ-DCAQKATOSA-N His-Ser-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CN=CN1 FHKZHRMERJUXRJ-DCAQKATOSA-N 0.000 description 1
- FRDFAWHTPDKRHG-ULQDDVLXSA-N His-Tyr-Arg Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)C1=CN=CN1 FRDFAWHTPDKRHG-ULQDDVLXSA-N 0.000 description 1
- 101001011382 Homo sapiens Interferon regulatory factor 3 Proteins 0.000 description 1
- 101100310115 Homo sapiens SETD5 gene Proteins 0.000 description 1
- 101000763537 Homo sapiens Toll-like receptor 10 Proteins 0.000 description 1
- 102000043138 IRF family Human genes 0.000 description 1
- YOTNPRLPIPHQSB-XUXIUFHCSA-N Ile-Arg-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N YOTNPRLPIPHQSB-XUXIUFHCSA-N 0.000 description 1
- BEWFWZRGBDVXRP-PEFMBERDSA-N Ile-Glu-Asn Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O BEWFWZRGBDVXRP-PEFMBERDSA-N 0.000 description 1
- KEKTTYCXKGBAAL-VGDYDELISA-N Ile-His-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CO)C(=O)O)N KEKTTYCXKGBAAL-VGDYDELISA-N 0.000 description 1
- HUORUFRRJHELPD-MNXVOIDGSA-N Ile-Leu-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N HUORUFRRJHELPD-MNXVOIDGSA-N 0.000 description 1
- ZLFNNVATRMCAKN-ZKWXMUAHSA-N Ile-Ser-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)NCC(=O)O)N ZLFNNVATRMCAKN-ZKWXMUAHSA-N 0.000 description 1
- HXIDVIFHRYRXLZ-NAKRPEOUSA-N Ile-Ser-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)O)N HXIDVIFHRYRXLZ-NAKRPEOUSA-N 0.000 description 1
- DGTOKVBDZXJHNZ-WZLNRYEVSA-N Ile-Thr-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N DGTOKVBDZXJHNZ-WZLNRYEVSA-N 0.000 description 1
- ZGKVPOSSTGHJAF-HJPIBITLSA-N Ile-Tyr-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CO)C(=O)O)N ZGKVPOSSTGHJAF-HJPIBITLSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 108010032038 Interferon Regulatory Factor-3 Proteins 0.000 description 1
- 102000002227 Interferon Type I Human genes 0.000 description 1
- 108010014726 Interferon Type I Proteins 0.000 description 1
- 102100029843 Interferon regulatory factor 3 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 108090000174 Interleukin-10 Proteins 0.000 description 1
- 101150068332 KIT gene Proteins 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
- 241000880493 Leptailurus serval Species 0.000 description 1
- WNGVUZWBXZKQES-YUMQZZPRSA-N Leu-Ala-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O WNGVUZWBXZKQES-YUMQZZPRSA-N 0.000 description 1
- HBJZFCIVFIBNSV-DCAQKATOSA-N Leu-Arg-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(N)=O)C(O)=O HBJZFCIVFIBNSV-DCAQKATOSA-N 0.000 description 1
- GRZSCTXVCDUIPO-SRVKXCTJSA-N Leu-Arg-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O GRZSCTXVCDUIPO-SRVKXCTJSA-N 0.000 description 1
- STAVRDQLZOTNKJ-RHYQMDGZSA-N Leu-Arg-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O STAVRDQLZOTNKJ-RHYQMDGZSA-N 0.000 description 1
- DLCOFDAHNMMQPP-SRVKXCTJSA-N Leu-Asp-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O DLCOFDAHNMMQPP-SRVKXCTJSA-N 0.000 description 1
- PVMPDMIKUVNOBD-CIUDSAMLSA-N Leu-Asp-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O PVMPDMIKUVNOBD-CIUDSAMLSA-N 0.000 description 1
- LOLUPZNNADDTAA-AVGNSLFASA-N Leu-Gln-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LOLUPZNNADDTAA-AVGNSLFASA-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
- LLBQJYDYOLIQAI-JYJNAYRXSA-N Leu-Glu-Tyr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O LLBQJYDYOLIQAI-JYJNAYRXSA-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
- XQXGNBFMAXWIGI-MXAVVETBSA-N Leu-His-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(C)C)CC1=CN=CN1 XQXGNBFMAXWIGI-MXAVVETBSA-N 0.000 description 1
- CSFVADKICPDRRF-KKUMJFAQSA-N Leu-His-Leu Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C([O-])=O)CC1=CN=CN1 CSFVADKICPDRRF-KKUMJFAQSA-N 0.000 description 1
- OHZIZVWQXJPBJS-IXOXFDKPSA-N Leu-His-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OHZIZVWQXJPBJS-IXOXFDKPSA-N 0.000 description 1
- KUIDCYNIEJBZBU-AJNGGQMLSA-N Leu-Ile-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O KUIDCYNIEJBZBU-AJNGGQMLSA-N 0.000 description 1
- DSFYPIUSAMSERP-IHRRRGAJSA-N Leu-Leu-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DSFYPIUSAMSERP-IHRRRGAJSA-N 0.000 description 1
- IAJFFZORSWOZPQ-SRVKXCTJSA-N Leu-Leu-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IAJFFZORSWOZPQ-SRVKXCTJSA-N 0.000 description 1
- 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
- VCHVSKNMTXWIIP-SRVKXCTJSA-N Leu-Lys-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O VCHVSKNMTXWIIP-SRVKXCTJSA-N 0.000 description 1
- LZHJZLHSRGWBBE-IHRRRGAJSA-N Leu-Lys-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O LZHJZLHSRGWBBE-IHRRRGAJSA-N 0.000 description 1
- HDHQQEDVWQGBEE-DCAQKATOSA-N Leu-Met-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(O)=O HDHQQEDVWQGBEE-DCAQKATOSA-N 0.000 description 1
- PTRKPHUGYULXPU-KKUMJFAQSA-N Leu-Phe-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O PTRKPHUGYULXPU-KKUMJFAQSA-N 0.000 description 1
- XWEVVRRSIOBJOO-SRVKXCTJSA-N Leu-Pro-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O XWEVVRRSIOBJOO-SRVKXCTJSA-N 0.000 description 1
- KZZCOWMDDXDKSS-CIUDSAMLSA-N Leu-Ser-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O KZZCOWMDDXDKSS-CIUDSAMLSA-N 0.000 description 1
- 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 1
- GOFJOGXGMPHOGL-DCAQKATOSA-N Leu-Ser-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(C)C GOFJOGXGMPHOGL-DCAQKATOSA-N 0.000 description 1
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 1
- LCNASHSOFMRYFO-WDCWCFNPSA-N Leu-Thr-Gln Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(N)=O LCNASHSOFMRYFO-WDCWCFNPSA-N 0.000 description 1
- ODRREERHVHMIPT-OEAJRASXSA-N Leu-Thr-Phe Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 ODRREERHVHMIPT-OEAJRASXSA-N 0.000 description 1
- BTEMNFBEAAOGBR-BZSNNMDCSA-N Leu-Tyr-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BTEMNFBEAAOGBR-BZSNNMDCSA-N 0.000 description 1
- YQFZRHYZLARWDY-IHRRRGAJSA-N Leu-Val-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN YQFZRHYZLARWDY-IHRRRGAJSA-N 0.000 description 1
- 108010028921 Lipopeptides Proteins 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- NTSPQIONFJUMJV-AVGNSLFASA-N Lys-Arg-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(O)=O NTSPQIONFJUMJV-AVGNSLFASA-N 0.000 description 1
- FACUGMGEFUEBTI-SRVKXCTJSA-N Lys-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCCCN FACUGMGEFUEBTI-SRVKXCTJSA-N 0.000 description 1
- RZHLIPMZXOEJTL-AVGNSLFASA-N Lys-Gln-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCCN)N RZHLIPMZXOEJTL-AVGNSLFASA-N 0.000 description 1
- MQMIRLVJXQNTRJ-SDDRHHMPSA-N Lys-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCCN)N)C(=O)O MQMIRLVJXQNTRJ-SDDRHHMPSA-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
- JOSAKOKSPXROGQ-BJDJZHNGSA-N Lys-Ser-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JOSAKOKSPXROGQ-BJDJZHNGSA-N 0.000 description 1
- RPWTZTBIFGENIA-VOAKCMCISA-N Lys-Thr-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O RPWTZTBIFGENIA-VOAKCMCISA-N 0.000 description 1
- PHKBGZKVOJCIMZ-SRVKXCTJSA-N Met-Pro-Arg Chemical compound CSCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O PHKBGZKVOJCIMZ-SRVKXCTJSA-N 0.000 description 1
- 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 1
- AYPMIIKUMNADSU-IHRRRGAJSA-N Phe-Arg-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O AYPMIIKUMNADSU-IHRRRGAJSA-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
- YZJKNDCEPDDIDA-BZSNNMDCSA-N Phe-His-Lys Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC=CC=1)C1=CN=CN1 YZJKNDCEPDDIDA-BZSNNMDCSA-N 0.000 description 1
- RGZYXNFHYRFNNS-MXAVVETBSA-N Phe-Ile-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N RGZYXNFHYRFNNS-MXAVVETBSA-N 0.000 description 1
- RORUIHAWOLADSH-HJWJTTGWSA-N Phe-Ile-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=CC=C1 RORUIHAWOLADSH-HJWJTTGWSA-N 0.000 description 1
- MSHZERMPZKCODG-ACRUOGEOSA-N Phe-Leu-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 MSHZERMPZKCODG-ACRUOGEOSA-N 0.000 description 1
- INHMISZWLJZQGH-ULQDDVLXSA-N Phe-Leu-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 INHMISZWLJZQGH-ULQDDVLXSA-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
- CZQZSMJXFGGBHM-KKUMJFAQSA-N Phe-Pro-Gln Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O CZQZSMJXFGGBHM-KKUMJFAQSA-N 0.000 description 1
- GMWNQSGWWGKTSF-LFSVMHDDSA-N Phe-Thr-Ala Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O GMWNQSGWWGKTSF-LFSVMHDDSA-N 0.000 description 1
- BSTPNLNKHKBONJ-HTUGSXCWSA-N Phe-Thr-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N)O BSTPNLNKHKBONJ-HTUGSXCWSA-N 0.000 description 1
- XNQMZHLAYFWSGJ-HTUGSXCWSA-N Phe-Thr-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O XNQMZHLAYFWSGJ-HTUGSXCWSA-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
- FRMKIPSIZSFTTE-HJOGWXRNSA-N Phe-Tyr-Phe 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](CC1=CC=CC=C1)C(O)=O FRMKIPSIZSFTTE-HJOGWXRNSA-N 0.000 description 1
- CYQQWUPHIZVCNY-GUBZILKMSA-N Pro-Arg-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O CYQQWUPHIZVCNY-GUBZILKMSA-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
- NOXSEHJOXCWRHK-DCAQKATOSA-N Pro-Cys-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@@H]1CCCN1 NOXSEHJOXCWRHK-DCAQKATOSA-N 0.000 description 1
- HJSCRFZVGXAGNG-SRVKXCTJSA-N Pro-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1 HJSCRFZVGXAGNG-SRVKXCTJSA-N 0.000 description 1
- LQZZPNDMYNZPFT-KKUMJFAQSA-N Pro-Gln-Phe Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LQZZPNDMYNZPFT-KKUMJFAQSA-N 0.000 description 1
- NMELOOXSGDRBRU-YUMQZZPRSA-N Pro-Glu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CCC(=O)O)NC(=O)[C@@H]1CCCN1 NMELOOXSGDRBRU-YUMQZZPRSA-N 0.000 description 1
- UEHYFUCOGHWASA-HJGDQZAQSA-N Pro-Glu-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CCCN1 UEHYFUCOGHWASA-HJGDQZAQSA-N 0.000 description 1
- UIMCLYYSUCIUJM-UWVGGRQHSA-N Pro-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 UIMCLYYSUCIUJM-UWVGGRQHSA-N 0.000 description 1
- HAEGAELAYWSUNC-WPRPVWTQSA-N Pro-Gly-Val Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O HAEGAELAYWSUNC-WPRPVWTQSA-N 0.000 description 1
- VTFXTWDFPTWNJY-RHYQMDGZSA-N Pro-Leu-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VTFXTWDFPTWNJY-RHYQMDGZSA-N 0.000 description 1
- WHNJMTHJGCEKGA-ULQDDVLXSA-N Pro-Phe-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O WHNJMTHJGCEKGA-ULQDDVLXSA-N 0.000 description 1
- NAIPAPCKKRCMBL-JYJNAYRXSA-N Pro-Pro-Phe Chemical compound C([C@@H](C(=O)O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H]1NCCC1)C1=CC=CC=C1 NAIPAPCKKRCMBL-JYJNAYRXSA-N 0.000 description 1
- RNEFESSBTOQSAC-DCAQKATOSA-N Pro-Ser-His Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O RNEFESSBTOQSAC-DCAQKATOSA-N 0.000 description 1
- DCHQYSOGURGJST-FJXKBIBVSA-N Pro-Thr-Gly Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O DCHQYSOGURGJST-FJXKBIBVSA-N 0.000 description 1
- KHRLUIPIMIQFGT-AVGNSLFASA-N Pro-Val-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O KHRLUIPIMIQFGT-AVGNSLFASA-N 0.000 description 1
- 208000005074 Retroviridae Infections Diseases 0.000 description 1
- 101150103943 SETD5 gene Proteins 0.000 description 1
- 108010044012 STAT1 Transcription Factor Proteins 0.000 description 1
- 102000006381 STAT1 Transcription Factor Human genes 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- GXXTUIUYTWGPMV-FXQIFTODSA-N Ser-Arg-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O GXXTUIUYTWGPMV-FXQIFTODSA-N 0.000 description 1
- QFBNNYNWKYKVJO-DCAQKATOSA-N Ser-Arg-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CCCN=C(N)N QFBNNYNWKYKVJO-DCAQKATOSA-N 0.000 description 1
- ULVMNZOKDBHKKI-ACZMJKKPSA-N Ser-Gln-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O ULVMNZOKDBHKKI-ACZMJKKPSA-N 0.000 description 1
- SMIDBHKWSYUBRZ-ACZMJKKPSA-N Ser-Glu-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O SMIDBHKWSYUBRZ-ACZMJKKPSA-N 0.000 description 1
- SQBLRDDJTUJDMV-ACZMJKKPSA-N Ser-Glu-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O SQBLRDDJTUJDMV-ACZMJKKPSA-N 0.000 description 1
- GYXVUTAOICLGKJ-ACZMJKKPSA-N Ser-Glu-Cys Chemical compound C(CC(=O)O)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CO)N GYXVUTAOICLGKJ-ACZMJKKPSA-N 0.000 description 1
- UOLGINIHBRIECN-FXQIFTODSA-N Ser-Glu-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O UOLGINIHBRIECN-FXQIFTODSA-N 0.000 description 1
- GZFAWAQTEYDKII-YUMQZZPRSA-N Ser-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO GZFAWAQTEYDKII-YUMQZZPRSA-N 0.000 description 1
- MOINZPRHJGTCHZ-MMWGEVLESA-N Ser-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N MOINZPRHJGTCHZ-MMWGEVLESA-N 0.000 description 1
- UIPXCLNLUUAMJU-JBDRJPRFSA-N Ser-Ile-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O UIPXCLNLUUAMJU-JBDRJPRFSA-N 0.000 description 1
- MQQBBLVOUUJKLH-HJPIBITLSA-N Ser-Ile-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O MQQBBLVOUUJKLH-HJPIBITLSA-N 0.000 description 1
- KCNSGAMPBPYUAI-CIUDSAMLSA-N Ser-Leu-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O KCNSGAMPBPYUAI-CIUDSAMLSA-N 0.000 description 1
- GVMUJUPXFQFBBZ-GUBZILKMSA-N Ser-Lys-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O GVMUJUPXFQFBBZ-GUBZILKMSA-N 0.000 description 1
- PMCMLDNPAZUYGI-DCAQKATOSA-N Ser-Lys-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O PMCMLDNPAZUYGI-DCAQKATOSA-N 0.000 description 1
- UPLYXVPQLJVWMM-KKUMJFAQSA-N Ser-Phe-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O UPLYXVPQLJVWMM-KKUMJFAQSA-N 0.000 description 1
- MQUZANJDFOQOBX-SRVKXCTJSA-N Ser-Phe-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O MQUZANJDFOQOBX-SRVKXCTJSA-N 0.000 description 1
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 1
- RXUOAOOZIWABBW-XGEHTFHBSA-N Ser-Thr-Arg Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N RXUOAOOZIWABBW-XGEHTFHBSA-N 0.000 description 1
- YEDSOSIKVUMIJE-DCAQKATOSA-N Ser-Val-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O YEDSOSIKVUMIJE-DCAQKATOSA-N 0.000 description 1
- LGIMRDKGABDMBN-DCAQKATOSA-N Ser-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N LGIMRDKGABDMBN-DCAQKATOSA-N 0.000 description 1
- HSWXBJCBYSWBPT-GUBZILKMSA-N Ser-Val-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)C(O)=O HSWXBJCBYSWBPT-GUBZILKMSA-N 0.000 description 1
- 108020004459 Small interfering RNA Proteins 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- NJEMRSFGDNECGF-GCJQMDKQSA-N Thr-Ala-Asp Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O NJEMRSFGDNECGF-GCJQMDKQSA-N 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
- NFMPFBCXABPALN-OWLDWWDNSA-N Thr-Ala-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N)O NFMPFBCXABPALN-OWLDWWDNSA-N 0.000 description 1
- GARULAKWZGFIKC-RWRJDSDZSA-N Thr-Gln-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GARULAKWZGFIKC-RWRJDSDZSA-N 0.000 description 1
- IGGFFPOIFHZYKC-PBCZWWQYSA-N Thr-His-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O IGGFFPOIFHZYKC-PBCZWWQYSA-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
- VTVVYQOXJCZVEB-WDCWCFNPSA-N Thr-Leu-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O VTVVYQOXJCZVEB-WDCWCFNPSA-N 0.000 description 1
- RFKVQLIXNVEOMB-WEDXCCLWSA-N Thr-Leu-Gly Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)O)N)O RFKVQLIXNVEOMB-WEDXCCLWSA-N 0.000 description 1
- VRUFCJZQDACGLH-UVOCVTCTSA-N Thr-Leu-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VRUFCJZQDACGLH-UVOCVTCTSA-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
- SOUPNXUJAJENFU-SWRJLBSHSA-N Thr-Trp-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O SOUPNXUJAJENFU-SWRJLBSHSA-N 0.000 description 1
- KPMIQCXJDVKWKO-IFFSRLJSSA-N Thr-Val-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KPMIQCXJDVKWKO-IFFSRLJSSA-N 0.000 description 1
- BKVICMPZWRNWOC-RHYQMDGZSA-N Thr-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)[C@@H](C)O BKVICMPZWRNWOC-RHYQMDGZSA-N 0.000 description 1
- 102100027009 Toll-like receptor 10 Human genes 0.000 description 1
- OQMQBYOEAHVCGD-GQGQLFGLSA-N Trp-Cys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N OQMQBYOEAHVCGD-GQGQLFGLSA-N 0.000 description 1
- DVIIYMVCSUQOJG-QEJZJMRPSA-N Trp-Glu-Asp Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O DVIIYMVCSUQOJG-QEJZJMRPSA-N 0.000 description 1
- CKKFTIQYURNSEI-IHRRRGAJSA-N Tyr-Asn-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 CKKFTIQYURNSEI-IHRRRGAJSA-N 0.000 description 1
- PMDWYLVWHRTJIW-STQMWFEESA-N Tyr-Gly-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 PMDWYLVWHRTJIW-STQMWFEESA-N 0.000 description 1
- QARCDOCCDOLJSF-HJPIBITLSA-N Tyr-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N QARCDOCCDOLJSF-HJPIBITLSA-N 0.000 description 1
- KSCVLGXNQXKUAR-JYJNAYRXSA-N Tyr-Leu-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KSCVLGXNQXKUAR-JYJNAYRXSA-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
- UUYCNAXCCDNULB-QXEWZRGKSA-N Val-Arg-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(N)=O)C(O)=O UUYCNAXCCDNULB-QXEWZRGKSA-N 0.000 description 1
- COYSIHFOCOMGCF-WPRPVWTQSA-N Val-Arg-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CCCN=C(N)N COYSIHFOCOMGCF-WPRPVWTQSA-N 0.000 description 1
- COYSIHFOCOMGCF-UHFFFAOYSA-N Val-Arg-Gly Natural products CC(C)C(N)C(=O)NC(C(=O)NCC(O)=O)CCCN=C(N)N COYSIHFOCOMGCF-UHFFFAOYSA-N 0.000 description 1
- PVPAOIGJYHVWBT-KKHAAJSZSA-N Val-Asn-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C(C)C)N)O PVPAOIGJYHVWBT-KKHAAJSZSA-N 0.000 description 1
- BWVHQINTNLVWGZ-ZKWXMUAHSA-N Val-Cys-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(=O)O)C(=O)O)N BWVHQINTNLVWGZ-ZKWXMUAHSA-N 0.000 description 1
- JXGWQYWDUOWQHA-DZKIICNBSA-N Val-Gln-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N JXGWQYWDUOWQHA-DZKIICNBSA-N 0.000 description 1
- FOADDSDHGRFUOC-DZKIICNBSA-N Val-Glu-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N FOADDSDHGRFUOC-DZKIICNBSA-N 0.000 description 1
- NXRAUQGGHPCJIB-RCOVLWMOSA-N Val-Gly-Asn Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O NXRAUQGGHPCJIB-RCOVLWMOSA-N 0.000 description 1
- OVBMCNDKCWAXMZ-NAKRPEOUSA-N Val-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](C(C)C)N OVBMCNDKCWAXMZ-NAKRPEOUSA-N 0.000 description 1
- DJQIUOKSNRBTSV-CYDGBPFRSA-N Val-Ile-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](C(C)C)N DJQIUOKSNRBTSV-CYDGBPFRSA-N 0.000 description 1
- OJPRSVJGNCAKQX-SRVKXCTJSA-N Val-Met-Arg Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N OJPRSVJGNCAKQX-SRVKXCTJSA-N 0.000 description 1
- KSFXWENSJABBFI-ZKWXMUAHSA-N Val-Ser-Asn Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O KSFXWENSJABBFI-ZKWXMUAHSA-N 0.000 description 1
- IRAUYEAFPFPVND-UVBJJODRSA-N Val-Trp-Ala Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](N)C(C)C)C(=O)N[C@@H](C)C(O)=O)=CNC2=C1 IRAUYEAFPFPVND-UVBJJODRSA-N 0.000 description 1
- LLJLBRRXKZTTRD-GUBZILKMSA-N Val-Val-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N LLJLBRRXKZTTRD-GUBZILKMSA-N 0.000 description 1
- 229920000392 Zymosan Polymers 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 210000005006 adaptive immune system Anatomy 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010050025 alpha-glutamyltryptophan Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000003975 animal breeding Methods 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 1
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000010523 cascade reaction Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000004163 cytometry Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 238000000432 density-gradient centrifugation Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 230000007783 downstream signaling Effects 0.000 description 1
- 210000001671 embryonic stem cell Anatomy 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 230000012173 estrus Effects 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010046775 glutamyl-isoleucyl-leucyl-aspartyl-valine Proteins 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 108010013726 glycyl-arginyl-glycyl-glutamyl-serine Proteins 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010010147 glycylglutamine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 108010053037 kyotorphin Proteins 0.000 description 1
- 125000001909 leucine group Chemical group [H]N(*)C(C(*)=O)C([H])([H])C(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000009437 off-target effect Effects 0.000 description 1
- 210000000287 oocyte Anatomy 0.000 description 1
- 210000003101 oviduct Anatomy 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 108010064486 phenylalanyl-leucyl-valine Proteins 0.000 description 1
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- 108010053725 prolylvaline Proteins 0.000 description 1
- 108010054624 red fluorescent protein Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 238000010374 somatic cell nuclear transfer Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 210000001635 urinary tract Anatomy 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 239000013603 viral vector Substances 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
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70596—Molecules with a "CD"-designation not provided for elsewhere
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
- A01K67/0278—Knock-in vertebrates, e.g. humanised vertebrates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/15—Humanized animals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/102—Caprine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/02—Animal zootechnically ameliorated
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
- C07K2319/41—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation containing a Myc-tag
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Veterinary Medicine (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Biomedical Technology (AREA)
- Environmental Sciences (AREA)
- Toxicology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Animal Behavior & Ethology (AREA)
- Plant Pathology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了嵌合抗病基因及其相关生物材料与应用。具体地公开了氨基酸序列是SEQ ID No.2的嵌合蛋白质TLR2‑4,包括TLR2的胞外区、TLR4的跨膜区与胞内区。本发明还提供了嵌合蛋白质TLR2‑4的编码基因,以及含有该编码基因的重组载体和重组细胞。本发明TLR2‑4基因在山羊成纤维细胞中转录成功表达,能够执行模式识别受体的功能产生更多的细胞因子和趋化因子,增强了山羊成纤维细胞在识别S.aureus后募集天然吞噬细胞的能力。本发明嵌合TLR2‑4基因能够促进巨噬细胞发挥吞噬作用,增强免疫力,提高山羊细胞对导致乳腺炎的常见病原菌金黄色葡萄球菌的抗性,提高山羊的抗病能力。
Description
技术领域
本发明属于基因工程技术及动物抗病育种领域,具体涉及嵌合抗病基因及其相关生物材料与应用。
背景技术
先天免疫系统是宿主抵抗外界微生物入侵的第一道防线,在机体识别和抵抗微生物入侵的过程中起到重要作用。在先天免疫系统中由模式识别受体(PRRs)负责识别入侵病原的分子模式(Pathogen-associated molecular pattern,PAMPs),Toll样受体(Tolllike receptors,TLRs)是PRRs中的重要家族,是一类广泛表达于哺乳动物细胞表面的跨膜信号转导蛋白,能够通过与相应PAMPs的结合触发信号通路中的级联反应,诱导炎症因子及细胞因子的产生,激活固有免疫系统(先天免疫系统)和获得性免疫系统,来共同抵抗病原微生物的入侵,在宿主抵抗病原感染中发挥重要作用。
TLRs是由胞外区、跨膜区和胞内区组成的Ⅰ型跨膜信号蛋白受体。TLRs的胞外区包含亮氨酸重复序列(Leucine-richrepeat,LRR),负责各种病原微生物的PAMPs的识别;跨膜区主要负责TLRs在细胞膜上的定位;胞内区由于与白介素1受体(IL-1R)具有同源性而被称为TIR(Toll-IL-1Receptor)结构域,该结构域通过募集下游含有TIR结构的信号分子从而介导信号级联反应,活化信号通路。现已发现的TLR家族成员有13个,其中TLR1-9在人类和鼠类是相同的,而TLR10是人类所特有的,TLR11-13则是鼠类特有的。TLRs可以表达于多种免疫细胞,如单核巨噬细胞、淋巴细胞、树突状细胞等,另外还可以表达于气道、泌尿道等上皮细胞。目前牛、猪、绵羊、山羊等家畜的TLRs基因已被确认,大多数哺乳动物含有10-12个TLRs,不同受体(TLR)可识别不同的分子模式(PAMP),已发现多个PAMP能被人类Toll样受体所识别,例如TLR4(第一个在人体发现的TLRs家族成员)能够通过形成同源二聚体识别革兰氏阴性菌细胞壁外层的脂多糖(LPS);TLR2的配体较TLR4的广泛,可识别脂多肽,脂蛋白,脂壁酸(LTA)、脂阿拉伯甘聚糖(LAM)及酵母多糖等多种分子。TLR4与其配体LPS结合后,可以使转录因子干扰素调节因子3(interferonregulatefactor3,IRF3)活化,IRF3活化后可诱导IFN-β表达,之后通过IFN-β激活STAT1,最后导致Ⅰ型干扰素及其炎症因子的表达,TLR4不需要与其他TLRs形成异源二聚体来发挥作用。而TLR2需要通过与TLR1或TLR6形成TLR2/TLR1或TLR2/TLR6异源二聚体后来识别和清除病原菌,发挥生物学功能。
发明内容
本发明所要解决的技术问题是如何增强动物免疫力和/或提高动物抗病能力。所要解决的技术问题不限于如所描述的技术主题,本领域技术人员通过以下描述可以清楚地理解本文未提及的其它技术主题。
为解决上述技术问题,本发明首先提供了嵌合蛋白质(融合蛋白质),名称为TLR2-4,所述嵌合蛋白质包括TLR2的胞外区和TLR4的跨膜区与胞内区。
进一步地,所述嵌合蛋白质还包括信号肽和/或标签蛋白。
本领域技术人员应当理解,在研究某一目的蛋白功能时,将一个通过各种方法可以检测到的标签蛋白的基因和目的蛋白的基因进行重组融合,产生的融合蛋白除了目的蛋白本身,同时会带上可检测的标签蛋白,这样目的蛋白也一并检测到,其目的在于能够定位目的蛋白或研究蛋白相互作用,因此,适用于本申请的标签蛋白并不局限于特定种类。
所述标签蛋白包括但不限于:GST(谷胱甘肽巯基转移酶)标签蛋白、His6标签蛋白(His-tag)、MBP(麦芽糖结合蛋白)标签蛋白、Flag标签蛋白、SUMO标签蛋白、HA标签蛋白、Myc标签蛋白、eGFP(增强型绿色荧光蛋白)、eCFP(增强型青色荧光蛋白)、eYFP(增强型黄绿色荧光蛋白)、mCherry(单体红色荧光蛋白)或AviTag标签蛋白。
进一步地,所述信号肽可为TLR2的信号肽和/或所述标签蛋白可为Myc。
进一步地,所述TLR2的胞外区为下述任一种蛋白质:
P1)氨基酸序列是SEQ ID No.2的第31-597位的蛋白质;
P2)将P1)所示的蛋白质经过氨基酸残基的取代和/或缺失和/或添加得到的与P1)所示的蛋白质具有80%以上的同一性且具有相同功能的蛋白质;
所述TLR4的跨膜区与胞内区为下述任一种蛋白质:
Q1)氨基酸序列是SEQ ID No.2的第598-805位的蛋白质;
Q2)将Q1)所示的蛋白质经过氨基酸残基的取代和/或缺失和/或添加得到的与Q1)所示的蛋白质具有80%以上的同一性且具有相同功能的蛋白质。
所述嵌合蛋白质具体可由所述TLR2的胞外区和所述TLR4的跨膜区与胞内区连接而成,也可由所述信号肽、所述TLR2的胞外区和所述TLR4的跨膜区与胞内区连接而成,也可由所述标签蛋白、所述TLR2的胞外区和所述TLR4的跨膜区与胞内区连接而成,也可由所述标签蛋白、所述信号肽、所述TLR2的胞外区和所述TLR4的跨膜区与胞内区连接而成。
进一步地,所述嵌合蛋白质可为下述任一种蛋白质:
A1)氨基酸序列是SEQ ID No.2的蛋白质;
A2)氨基酸序列是SEQ ID No.2的第31-805位的蛋白质;
A3)将SEQ ID No.2的氨基酸序列经过氨基酸残基的取代和/或缺失和/或添加得到的与A1)所示的蛋白质具有80%以上的同一性且具有相同功能的蛋白质;
A4)将SEQ ID No.2的第31-805位的氨基酸序列经过氨基酸残基的取代和/或缺失和/或添加得到的与A2)所示的蛋白质具有80%以上的同一性且具有相同功能的蛋白质。
SEQ ID No.1所示的DNA分子编码SEQ ID No.2所示的TLR2-4嵌合蛋白质。
SEQ ID No.1的第91-2418位所示的DNA分子编码SEQ ID No.2的第31-805位所示的TLR2-4嵌合蛋白质。
其中,
SEQ ID No.2的第1-20位是TLR2的信号肽的氨基酸序列;
SEQ ID No.2的第21-30位是标签蛋白Myc的氨基酸序列;
SEQ ID No.2的第31-597位是TLR2的胞外区的氨基酸序列;
SEQ ID No.2的第598-805位是TLR4的跨膜区与胞内区的氨基酸序列;
SEQ ID No.1的第1-60位是TLR2的信号肽的核苷酸序列;
SEQ ID No.1的第61-90位是标签蛋白Myc的核苷酸序列;
SEQ ID No.1的第91-1791位是TLR2的胞外区的核苷酸序列;
SEQ ID No.1的第1792-2418位是TLR4的跨膜区与胞内区的核苷酸序列。
本领域普通技术人员可以很容易地采用已知的方法,例如定向进化或点突变的方法,对本发明的编码TLR2-4嵌合蛋白质的核苷酸序列进行突变。那些经过人工修饰的,具有与本发明分离得到的TLR2-4嵌合蛋白质的核苷酸序列75%或者更高同一性的核苷酸,只要编码TLR2-4嵌合蛋白质且具有TLR2-4嵌合蛋白质功能,均是衍生于本发明的核苷酸序列并且等同于本发明的序列。
上述75%或75%以上同一性,可为80%、85%、90%或95%以上的同一性。
本文中,同一性是指氨基酸序列或核苷酸序列的同一性。可使用国际互联网上的同源性检索站点测定氨基酸序列的同一性,如NCBI主页网站的BLAST网页。例如,可在高级BLAST2.1中,通过使用blastp作为程序,将Expect值设置为10,将所有Filter设置为OFF,使用BLOSUM62作为Matrix,将Gap existence cost,Per residue gap cost和Lambda ratio分别设置为11,1和0.85(缺省值)并进行检索一对氨基酸序列的同一性进行计算,然后即可获得同一性的值(%)。
本文中,所述80%以上的同一性可为至少80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%的同一性。
本发明还提供了生物材料,所述生物材料可为下述任一种:
B1)编码所述TLR2-4嵌合蛋白质的核酸分子;
B2)含有B1)所述核酸分子的表达盒;
B3)含有B1)所述核酸分子的重组载体、或含有B2)所述表达盒的重组载体;
B4)含有B1)所述核酸分子的重组微生物、或含有B2)所述表达盒的重组微生物、或含有B3)所述重组载体的重组微生物;
B5)含有B1)所述核酸分子的重组细胞、或含有B2)所述表达盒的重组细胞、或含有B3)所述重组载体的重组细胞。
所述载体可为质粒、黏粒、噬菌体或病毒载体。
所述微生物可为酵母、细菌、藻或真菌。其中,细菌可来自埃希氏菌属(Escherichia),欧文氏菌(Erwinia),根癌农杆菌属(Agrobacterium)、黄杆菌属(Flavobacterium),产碱菌属(Alcaligenes),假单胞菌属(Pseudomonas),芽胞杆菌属(Bacillus)等。
所述细胞可为动物细胞,具体可为山羊成纤维细胞。所述重组细胞具体可为转基因细胞(TLR2-4),所述转基因细胞(TLR2-4)可为含有TLR2-4基因(SEQ ID No.1)的重组山羊成纤维细胞。
上述生物材料中,所述核酸分子可为下述任一种:
C1)编码序列是SEQ ID No.1的DNA分子;
C2)编码序列是SEQ ID No.1的第91-2418位的DNA分子;
C3)与C1)或C2)限定的核苷酸序列具有75%或75%以上同一性,且编码所述嵌合蛋白质的DNA分子。
编码所述TLR2-4嵌合蛋白质的核酸分子可为TLR2-4嵌合基因(又称TLR2-4融合基因,简称TLR2-4基因),TLR2-4基因的核苷酸序列可为SEQ ID No.1。
上述生物材料中,所述重组载体还可包括CMV-tdtomato元件。
所述重组载体具体可为广谱表达TLR2-4的基因定点整合供体质粒pR-T2/4;pR-T2/4是以pRosa26-promoter为骨架载体,在Poly(A)上游引入EcoRI、PmlI酶切位点,上下游分别添加BamHI及XbaI酶切位点;用BamHI及XbaI同时双酶切pRosa26-promoter载体和poly(A),将poly(A)克隆到pRosa26-promoter载体;用XbaI和ScaI-HF双酶切,通过无缝克隆将右同源臂HA-R插入poly(A)后;用Acc65I和ScalI双酶切载体,在pRosa26-promoter载体上游引入PacI酶切位点并插入左同源臂HA-L,得到重组载体pRosa26-HA。pRosa26-HA是将pRosa26-promoter的限制性核酸内切酶BamHI和Xbal之间的小片段替换为核苷酸序列是SEQ ID No.3所示的DNA片段,保持pRosa26-promoter的其它核苷酸不变得到的重组载体。用PmlI和EcoRI-HF双酶切pRosa26-HA,在pRosa26-promoter与poly(A)之间插入TLR2-4基因(SEQ ID No.1),得到重组载体pRosa26-TLR2-4;用SacI-HF单酶切pRosa26-TLR2-4,在HA-R下游插入从pCMV-tdTomato载体中获得的CMV-tdtomato,获得完整的pRosa26-TLR2-4供体质粒,简称为pR-T2/4(图4)。
本发明首次构建了pR-T2/4广谱表达TLR2-4的基因定点整合供体质粒,并通过CRISPER/Cas9基因编辑系统将其导入山羊成纤维细胞中。进一步研究嵌合TLR2-4抗病基因表达的转基因细胞和野生对照细胞在TLR2配体和金黄色葡萄球菌攻毒后的抗病反应。
所述嵌合TLR2-4抗病基因能显著提高山羊成纤维细胞对金黄色葡萄球菌(Staphylococcus aureus)的抗性,增加细胞因子和趋化因子的分泌。
本发明还提供了一种培育抗病动物的方法,所述方法包括将编码TLR2-4嵌合蛋白质的核酸分子或所述重组载体导入受体动物。
进一步地,所述导入可为通过显微注射法、逆转录病毒感染法、胚胎干细胞介导法、体细胞核移植法、精子载体介导法或受体介导法等,将所述编码TLR2-4嵌合蛋白质的核酸分子或所述重组载体导入受体动物。
本发明还提供了应用,所述应用为M1或M2:
M1、所述生物材料在制备增强动物巨噬细胞对金黄色葡萄球菌的吞噬能力的产品中的应用,
M2、所述生物材料在制备增强动物细胞对金黄色葡萄球菌的抗性的产品中的应用。
本文中,所述动物可为山羊。所述山羊可为崂山羊。
所述抗病动物可为抗金黄色葡萄球菌动物,具体可为抗金黄色葡萄球菌山羊。
本发明还提供了TLR2-4嵌合蛋白质和/或所述生物材料在制备增强动物免疫力和/或提高动物抗病能力的产品中应用,和/或,在培育抗病动物中的应用,和/或,在动物育种中的应用。
本发明将山羊TLR2基因的胞外区序列与TLR4基因跨膜区和胞内区融合为TLR2-4嵌合基因,构建了广谱表达TLR2-4的基因定点整合供体质粒pR-T2/4。将供体质粒与PX458-2基因打靶载体共转染至山羊成纤维细胞,通过PCR鉴定筛选发生定点整合的细胞株。
实验证明,本发明与现有技术相比,具有以下优点:
(1)本发明构建的嵌合TLR2-4基因在山羊成纤维细胞中转录成功表达,而且能够执行模式识别受体的功能并比野生型细胞(WT)产生更多的细胞因子和趋化因子,增强了山羊成纤维细胞在识别S.aureus后募集天然吞噬细胞的能力。
(2)本发明构建的嵌合TLR2-4基因可增强巨噬细胞在识别S.aureus后产生的抗菌炎症反应。
(3)本发明构建的嵌合TLR2-4基因可促进巨噬细胞发挥吞噬作用,增强免疫力。
(4)本发明的嵌合TLR2-4抗病基因在山羊细胞中的表达能够提高山羊细胞对导致乳腺炎的常见病原菌金黄色葡萄球菌的抗性,提高了山羊的抗病能力。
附图说明
图1为sgRNA-Cas9载体结构图。
图2为sgRNA打靶示意图。
图3为Overlap PCR构建TLR2-4嵌合基因示意图。
图4为供体质粒pRosa26-TLR2-4(pR-T2/4)载体结构示意图。
图5为培养14天的山羊成纤维细胞单克隆(100×)。
图6为CRISPR/Cas9系统介导的TLR2-4嵌合基因定点整合鉴定结果。
图7为siTLR2对内源TLR2的沉默效率检测结果。
图8为TLR2-4对配体及病原菌的识别结果。
图9为TLR2-4巨噬细胞成功表达TLR2-4蛋白的鉴定结果图。
图10为TLR2-4巨噬细胞对金黄色葡萄球菌(S.aureus)的识别。
图11为TLR2-4巨噬细胞和WT巨噬细胞对金黄色葡萄球菌(S.aureus)的吞噬指数。
图12为TLR2-4巨噬细胞和WT巨噬细胞对金黄色葡萄球菌(S.aureus)的吞噬率。
具体实施方式
下面结合具体实施方式对本发明进行进一步的详细描述,给出的实施例仅为了阐明本发明,而不是为了限制本发明的范围。以下提供的实施例可作为本技术领域普通技术人员进行进一步改进的指南,并不以任何方式构成对本发明的限制。
下述实施例中的实验方法,如无特殊说明,均为常规方法,按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。
下述实施例中的PX458载体、pRosa26-promoter载体购于Addgene公司;pCMV-tdTomato载体购于Clontech公司;限制性核酸内切酶、T7E1酶、T4连接酶及Buffe r购于NEB公司;引物合成及序列测定由苏州金唯智公司完成;无缝克隆试剂盒Gene Art GibsonAssembly购于Thermo Fisher公司。
实施例1TLR2-4嵌合基因转染羊成纤维细胞
1、设计和筛选CRISPR/Cas9基因编辑靶点
将SETD5基因的内含子区作为基因编辑靶点筛选区设计基因编辑靶点。在该区域的DNA双链上寻找PAM区(5’-NGG),并将其5’端的20nt作为靶点序列。随后,选取PAM区5’端的13nt作为种子序列,与NCBI在线数据库进行比对以进行脱靶效应的估计,剔除掉除靶点外还有较多脱靶位点的靶点序列。使用sgRNA结构在线预测网站(http://unafold.rna.albany.edu/q=mfold/RNA-Folding-Form/)评估各靶点对应的sgRNA的结构及自由能,筛选出8个CRISPR/Cas9基因编辑靶点并进行细胞水平的验证。
使用BbsI核酸内切酶酶切PX458载体(购于Addgene公司),37℃酶切1h后通过1%琼脂糖凝胶电泳鉴定,并纯化回收酶切后的载体。将8组crRNA-oligo退火形成双链DNA后与纯化后的PX458载体连接。构建8个靶点筛选载体sgRNA-Cas9,结构如图1。
利用Amaxa’s Nucleofector电转仪(Lonza),程序:N-024,将8个靶点筛选载体sgRNA-Cas9分别转染至山羊成纤维细胞中。通过T7E1酶切(购于NEB公司)评估各靶点的基因编辑效率,综合打靶效率评估,选择靶点2作为CRISPR/Cas9的打靶序列。所构建的打靶载体为sgRNA2-Cas9打靶载体(也称为PX458-2基因打靶载体)。靶点2的相对位置及序列如图2所示。
sgRNA2-Cas9打靶载体的具体构建方法如下:将合成的引物SETD5-IN-sgRNA2-F(5’-caccgACTACACATTATAGATGACT-3’)和引物SETD5-IN-sgRNA2-R(5’-aaacAGTCATCTATAATGTGTAGTc-3’)用ddH2O稀释成100μM的溶液,在退火体系中添加SETD5-IN-sgRNA2-F和SETD5-IN-sgRNA2-R引物溶液各1μL,T4连接酶Buffer 1μL,ddH2O 7μL,混匀。将上述退火体系放入PCR仪中:95℃,2min;以-5℃/s的速度降至25℃,随后置于冰上,令sgRNA退火体系中的F和R引物互补配对,形成含有黏性末端的双链寡核苷酸(oligo);并取1μL oligo,用ddH2O稀释100倍,得到oligo稀释液。
将PX458载体用限制性内切酶BbsI酶切并回收纯化,得到酶切后的线性骨架载体,并在连接体系中添加100ng酶切后的线性骨架载体;在连接体系中加入1μL上述oligo稀释液、1μL T4连接酶Buffer、1μL T4连接酶,并用ddH2O补足至10μL,混匀,置于16℃过夜反应。通过常规转化法进行转化、涂板。待单菌落长成后,挑取数个扩大培养并测序验证,结果显示sgRNA识别区的编码DNA已成功连入骨架载体,得到sgRNA2-Cas9打靶载体。
2、构建TLR2-4嵌合基因
为了构建TLR2-4嵌合基因,提取山羊血液总RNA进行反转录,利用表1的引物进行山羊TLR2、TLR4基因外显子的扩增。
表1 PCR引物
利用表2的引物进行山羊TLR2基因胞外区外显子的扩增。PCR模板为上述获得的TLR2基因外显子PCR产物。
表2 PCR引物
利用表3的引物,在TLR2基因外显子的5’端添加信号肽及Myc序列,同时在TLR2基因胞外区外显子的3’端添加20bp的TLR4基因胞外区同源序列。
表3 PCR引物
将上述获得的TLR2基因胞外区外显子序列克隆至平末端载体。利用表4的引物进行Overlap PCR。
表4 PCR引物
通过Overlap PCR获得的信号肽-Myc-TLR2-4嵌合基因(以下简称TLR2-4基因)如图3所示,嵌合基因PCR产物为2418bp(SEQ ID No.1)。序列为:
TLR2-4嵌合基因的核苷酸序列为SEQ ID No.1,编码的嵌合蛋白TLR2-4的氨基酸序列为SEQ ID No.2。
SEQ ID No.1的第91-2418位是TLR2的胞外区及TLR4的跨膜区与胞内区的核苷酸序列;
SEQ ID No.2的第31-805位是TLR2的胞外区及TLR4的跨膜区与胞内区的氨基酸序列;
SEQ ID No.2的第1-20位是TLR2的信号肽的氨基酸序列;
SEQ ID No.2的第21-30位是标签蛋白Myc的氨基酸序列;
SEQ ID No.2的第31-597位是TLR2的胞外区的氨基酸序列;
SEQ ID No.2的第598-805位是TLR4的跨膜区与胞内区的氨基酸序列;
SEQ ID No.1的第1-60位是TLR2的信号肽的核苷酸序列;
SEQ ID No.1的第61-90位是标签蛋白Myc的核苷酸序列;
SEQ ID No.1的第91-1791位是TLR2的胞外区的核苷酸序列;
SEQ ID No.1的第1792-2418位是TLR4的跨膜区与胞内区的核苷酸序列。
3、构建供体质粒pR-T2/4
为后续的无缝克隆试验,需要在获得的TLR2-4嵌合基因两侧添加同源序列。以山羊基因组DNA为模板,利用表5的引物扩增左右同源臂(HA-L和HA-R)。
表5 PCR引物
通过表6的引物获得供体质粒pR-T2/4中所需元件CMV-tdtomato和poly(A)。
表6 PCR引物
表中pRosa26-promoter载体购自Addgene公司。
以pRosa26-promoter为骨架载体,在Poly(A)上游引入EcoRI、PmlI酶切位点,上下游分别添加BamHI及XbaI酶切位点;用BamHI及XbaI同时双酶切pRosa26-promoter载体和poly(A),将poly(A)克隆到pRosa26-promoter载体;用XbaI和ScaI-HF双酶切,通过无缝克隆将右同源臂HA-R插入Poly(A)后;用Acc65I和ScalI双酶切载体,在pRosa26-promoter载体上游引入PacI酶切位点并插入左同源臂HA-L,得到重组载体pRosa26-HA。pRosa26-HA是将pRosa26-promoter的限制性核酸内切酶BamHI和Xbal之间的小片段替换为核苷酸序列是SEQ ID No.3所示的DNA片段,保持pRosa26-promoter的其它核苷酸不变得到的重组载体。
用PmlI和EcoRI-HF双酶切pRosa26-HA,在pRosa26-promoter与poly(A)之间插入TLR2-4基因(SEQ ID No.1),得到重组载体pRosa26-TLR2-4;用SacI-HF单酶切pRosa26-TLR2-4,在HA-R下游插入从pCMV-tdTomato载体中获得的CMV-tdtomato,获得完整的pRosa26-TLR2-4供体质粒,简称为pR-T2/4(图4)。
4、细胞转染得到转基因细胞(TLR2-4)
将完整的pRosa26-TLR2-4供体质粒与sgRNA2-Cas9打靶载体(PX458-2基因打靶载体)共转染入崂山羊成纤维细胞(即野生型,WT)。消化转染培养3天后的成纤维细胞至流式管中,通过流式分选筛选出同时表达GFP及tdtomato的细胞。将分选获得的细胞以500个细胞/100mm培养皿的密度铺板培养10-14天,使用克隆环挑取细胞单克隆至96孔板中培养,获得的细胞单克隆如图5所示,得到转基因细胞(TLR2-4)。
扩大培养转基因细胞(TLR2-4),将上述获得的细胞单克隆裂解液作为模板,利用引物R-U和R-L进行PCR验证。
表7 PCR引物
结果显示,当存在定点整合的细胞时,可见1303bp的条带,而未转染的山羊成纤维细胞则无1303bp的条带,编号为1-C5的细胞单克隆利用引物R-U和R-L检出基因定点整合特异性条带,得到大小为1303bp的PCR产物,已成功定点整合了TLR2-4嵌合基因(图6)。图6中,各个泳道分别为1kb:1kb ladder;M3:Marker3;1-D3,1-C9,1-C5,1-C4,1-C2,1-B2,1-E3,1-D7为细胞单克隆号。
实施例2TLR2-4对配体及病原菌的识别
为了进一步检测TLR2-4能否形成二聚体并识别金黄色葡萄球菌及配体,通过qRT-PCR方法进行检测。为了排除细胞中内源TLR2的干扰并利于研究两组细胞的功能差异,使用siRNA沉默了内源TLR2基因。将siTLR2(sense 5’GCACUUCAACCCUCCCUUUTT3’;antisense 5’AAAGGGAGGGUUGAAGUGCTT3’)转染入转基因细胞(TLR2-4),得到重组细胞TLR2-4-siTLR2;将siTLR2转染入山羊成纤维细胞,得到重组细胞WTsiTLR2。转染24小时后,对重组细胞TLR2-4-siTLR2和重组细胞WTsiTLR2分别进行如下三种处理:1)加入500ng/mL的TLR2激动剂P2CSK4(P2C)溶液(购自InvivoGen公司,溶剂为无菌去离子水),该处理记为“TLR2-4-siTLR2+P2C”和“WTsiTLR2+P2C”;2)加入500ng/mL的P3CSK4(P3C)溶液(购自InvivoGen公司,溶剂为无菌去离子水),该处理记为“TLR2-4-siTLR2+P3C”和“WTsiTLR2+P3C”;3)加入MOI=10的热灭活金黄色葡萄球菌(S.aureus ATCC29213)(购自上海北诺生物科技有限公司)悬液(用无菌水重悬热灭活金黄色葡萄球菌(S.aureus)得到的液体),该处理记为“TLR2-4-siTLR2+S.aureus”和“WTsiTLR2+S.aureus”。qRT-PCR检测各处理内源TLR2基因的沉默情况,结果显示,通过siTLR2可以使细胞中85%以上的内源TLR2沉默(图7,“siTLR2组”表示重组细胞TLR2-4-siTLR2,“对照组”表示转基因细胞(TLR2-4))。选择高沉默效率的细胞,即重组细胞(TLR2-4-siTLR2)和野生型细胞(WT-siTLR2)通过qRT-PCR进行细胞因子IL-6、IL-8和IL-1β的转录情况的比较。
表8 qPCR引物
结果显示,转基因细胞(TLR2-4)识别配体P3C、P2C后产生比野生型细胞(WT)更多的细胞因子IL-6,识别金黄色葡萄球菌后产生比野生型细胞(WT)更多的细胞因子IL-8和IL6(图8)。采用Origin 8统计软件对数据进行处理,实验结果以平均值±标准偏差表示,采用Student’s t检验,P<0.001(***)表示具有极显著性差异。
结果表明本发明构建的嵌合TLR2-4基因在山羊成纤维细胞中转录成功表达,而且能够执行模式识别受体的功能并比野生型细胞(WT)产生更多的细胞因子和趋化因子,增强了山羊成纤维细胞在识别S.aureus后募集天然吞噬细胞的能力。
实施例3巨噬细胞的TLR2-4嵌合基因表达及对金黄色葡萄球菌的识别和吞噬能力的验证
1、巨噬细胞的TLR2-4嵌合基因表达
1-1核移植获得TLR2-4巨噬细胞
将实施例1步骤4中的阳性转基因细胞(TLR2-4)转移至100mm细胞培养皿中扩大培养,90%汇合后1-3d用于体细胞核移植。将转基因细胞(TLR2-4)移入去核的黑山羊的卵母细胞中,得到TLR2-4转基因克隆胚胎。126枚TLR2-4转基因克隆胚胎移植到12头同期发情的受体山羊(黑山羊)输卵管,经过妊娠、分娩,获得TLR2-4转基因山羊。采集山羊血液,通过密度梯度离心法分离获得TLR2-4转基因山羊和2只野生型山羊(崂山羊)外周血来源巨噬细胞(WT)。
1-2 Western blot检测巨噬细胞的TLR2-4嵌合基因表达
通过Western blot的方法使用Myc标签抗体检测巨噬细胞是否表达TLR2-4嵌合蛋白,结果如图9所示,TLR2-4巨噬细胞成功表达TLR2-4嵌合蛋白,对照组巨噬细胞(WT1和WT2)不表达TLR2-4蛋白。
2、TLR2-4巨噬细胞对金黄色葡萄球菌的识别
为了检测TLR2-4巨噬细胞能否识别金黄色葡萄球菌(S.aureus),首先用活的金黄色葡萄球菌(S.aureus ATCC29213)(购自上海北诺生物科技有限公司)分别感染TLR2-4巨噬细胞、WT巨噬细胞1小时,然后通过qRT-PCR对两组细胞的细胞因子表达情况进行检测。
表9 qPCR引物
结果显示,TLR2-4巨噬细胞识别金黄色葡萄球菌后产生比WT巨噬细胞更多的细胞因子IL-6、IL-8和IL-10(图10)。采用Origin 8统计软件对数据进行处理,实验结果以平均值±标准偏差表示,采用Student’s t检验,P<0.05(*)表示具有显著性差异,P<0.01(**)表示具有显著性差异,P<0.001(***)表示具有极显著性差异。
实验结果表明TLR2-4巨噬细胞表达的TLR2-4嵌合蛋白能够执行模式识别受体的功能,在识别S.aureus后产生比WT巨噬细胞更多的细胞因子和趋化因子,增强了山羊巨噬细胞在识别S.aureus后产生的抗菌炎症反应。
3、TLR2-4巨噬细胞对金黄色葡萄球菌的吞噬
为了检测TLR2-4巨噬细胞吞噬金黄色葡萄球菌的能力,使用FITC染色的金黄色葡萄球菌(S.aureus ATCC29213)悬液(MOI=1)感染TLR2-4巨噬细胞和WT巨噬细胞不同时间。PBS洗涤三次后,用含有200μg/ml庆大霉素的DMEM孵育细胞1小时以清除黏附在细胞表面的细菌。通过流式细胞术检测两组巨噬细胞的吞噬指数,即巨噬细胞中FITC的平均荧光强度(图11)。另外还检测了两组巨噬细胞的吞噬率,即FITC阳性细胞数/总细胞术×100%(图12)。结果显示TLR2-4巨噬细胞在15min,30min,60min,120min时对金黄色葡萄球菌的吞噬指数和吞噬率均高于WT巨噬细胞。
实验结果表明,在感染金黄色葡萄球菌后,表达嵌合蛋白TLR2-4的巨噬细胞发挥吞噬作用的细胞比例显著高于对照组(WT组),且TLR2-4巨噬细胞对金葡菌的吞噬能力显著高于WT组巨噬细胞。
本发明的嵌合TLR2-4抗病基因在山羊细胞中的表达能够提高山羊细胞对导致乳腺炎的常见病原菌金黄色葡萄球菌的抗性。
以上对本发明进行了详述。对于本领域技术人员来说,在不脱离本发明的宗旨和范围,以及无需进行不必要的实验情况下,可在等同参数、浓度和条件下,在较宽范围内实施本发明。虽然本发明给出了特殊的实施例,应该理解为,可以对本发明作进一步的改进。总之,按本发明的原理,本申请欲包括任何变更、用途或对本发明的改进,包括脱离了本申请中已公开范围,而用本领域已知的常规技术进行的改变。按以下附带的权利要求的范围,可以进行一些基本特征的应用。
SEQUENCE LISTING
<110> 中国农业大学
<120> 嵌合抗病基因及其相关生物材料与应用
<160> 3
<170> PatentIn version 3.5
<210> 1
<211> 2418
<212> DNA
<213> 人工序列(Artificial sequence)
<400> 1
atgccacgtg ctttgtggac agcgtgggtc tgggctgtaa tcagcgtgtt cacggaagga 60
gagcagaaac tcatctctga agaggatctg gcctctgatc aggcttcttc tctgtcttgt 120
gacccaactg gtgtctgcga tggccattcc agatctttaa actccatccc ctctggtctc 180
acagcaggtg tgaaaagcct tgacctgtcc gacaacgaga tcacctatgt cggcaacaga 240
gacctgcaga ggtgtgtgaa cctgaagact ctgaggctgg gggccaatga aattcacacg 300
gtggaggaag attctttttt tcacctgagg aatcttgaat atttggactt atcctataat 360
cgcttatcta acttatcatc ttcctggttc aggtcccttt acgtcttgaa attcttaaac 420
ttactgggaa atttatacaa aacacttggg gaaacatctc ttttttctca tctcccaaat 480
ctgcggaccc tgaaagtagg aaatagtaac agcttcactc agattcatga aaaggatttc 540
actggactga cttttcttga ggagcttgag atcagtgctc aaaatctgca gttatatgtg 600
ccaaagagtt taaagtcgat ccagaacatt agccatctga ttcttcatct gaagcagcct 660
gttttactcc tggacattct tatagatatt gtaagttcct tagattattt agaactgaga 720
gatactaatt tgcacacttt ctatttttca gaagcatcca tcagtgaagt taatacatca 780
gttaaaaagc ttatatttag aaatgtgcaa ttcaccgatg aaagttttgt tgaagttgtc 840
aaactgttta actatgtttc tgggatcttg gaagtagagt ttgatgactg tacccatgat 900
ggaattggcg attttacagc actgactttg aacagaatta gatacctagg taacgtggag 960
acgttaacaa tacggaagtt gcatatccca cagtttttct tattttatga tctgagtagt 1020
atatatccac tcacaggcaa agttaaaaga gtcacaatag aaaacagtaa ggttttcctg 1080
gttccttgtt tactttcaca acatttaaaa tcactagaat atttggatct cagtgaaaac 1140
ttaatgtctg aagaaacctt gagaaactca gcctgtgagc atgcctggcc cttccttcaa 1200
accctggttt taaggcagaa tcgtttgaaa tcactagaaa aaactggaga acttttgctt 1260
actctgaaaa atctgaataa ccttgatatc agtaagaata attttctttc aatgcctgaa 1320
acttgtcagt ggccaggaaa aatgaaacag ttgaacttat ccagcacgag gatacacagt 1380
ttaacccagt gccttcccca gaccctggaa attttagatg ttagcaataa caatctcgat 1440
tcattttctt tgattttgcc gcaactcaaa gaactttata tttccagaaa taagttgaag 1500
actctaccag atgcctcctt cttacccgtg ttatcagtta tgagaattag cggaaatata 1560
ataaatactt tctcgaagga acaacttgat tcttttccac aactgaaggc tttggaggcc 1620
ggtggcaaca acttcatttg ctcctgtgac ttcctgtcct tcacacaggg acagcaggca 1680
ctggcccgtg tcctggtcga ctggccagat ggctaccgct gtgacgctcc ctcgcacgtg 1740
cggggccagc gggtgcagga cgcccggctc tccctttctg aatgccaccg gatcatcagc 1800
gtgtcggttg tcactgtact cctggtatct gtggtaggag tcctagtcta taagttctat 1860
ttccacctga tgcttcttgc tggctgcaaa aagtatggca gaggtgaaag cacctatgat 1920
gcctttgtaa tctactcgag ccaggatgaa gcctgggtgc ggaatgaact ggtaaagaac 1980
ttggaggagg gcgtgccccc ctttcagctc tgccttcact acagggactt tattcctggg 2040
gtggccatcg ccgccaacat catccaggaa ggtttccaca agagccgtaa ggtgattgtc 2100
gtggtgtccc agcacttcat ccagagccga tggtgtatct tcgagtatga gattgcccag 2160
acctggcagt ttctgagcag ccgtgctggc atcatcttca tcgtcctgca gaagctggag 2220
aagtctctcc tgcggcagca ggtggaactc tatcgccttc tgaacaggaa cacctacctg 2280
gagtgggagg acagtgtcct ggggcggcat gtcttctgga gaagactcag aaaagccttg 2340
ctggctggta aaccccggag tccagaagga acagcagatg cagagaccaa cccgcaggaa 2400
gcgaccacct ccacctga 2418
<210> 2
<211> 805
<212> PRT
<213> 人工序列(Artificial sequence)
<400> 2
Met Pro Arg Ala Leu Trp Thr Ala Trp Val Trp Ala Val Ile Ser Val
1 5 10 15
Phe Thr Glu Gly Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Ala Ser
20 25 30
Asp Gln Ala Ser Ser Leu Ser Cys Asp Pro Thr Gly Val Cys Asp Gly
35 40 45
His Ser Arg Ser Leu Asn Ser Ile Pro Ser Gly Leu Thr Ala Gly Val
50 55 60
Lys Ser Leu Asp Leu Ser Asp Asn Glu Ile Thr Tyr Val Gly Asn Arg
65 70 75 80
Asp Leu Gln Arg Cys Val Asn Leu Lys Thr Leu Arg Leu Gly Ala Asn
85 90 95
Glu Ile His Thr Val Glu Glu Asp Ser Phe Phe His Leu Arg Asn Leu
100 105 110
Glu Tyr Leu Asp Leu Ser Tyr Asn Arg Leu Ser Asn Leu Ser Ser Ser
115 120 125
Trp Phe Arg Ser Leu Tyr Val Leu Lys Phe Leu Asn Leu Leu Gly Asn
130 135 140
Leu Tyr Lys Thr Leu Gly Glu Thr Ser Leu Phe Ser His Leu Pro Asn
145 150 155 160
Leu Arg Thr Leu Lys Val Gly Asn Ser Asn Ser Phe Thr Gln Ile His
165 170 175
Glu Lys Asp Phe Thr Gly Leu Thr Phe Leu Glu Glu Leu Glu Ile Ser
180 185 190
Ala Gln Asn Leu Gln Leu Tyr Val Pro Lys Ser Leu Lys Ser Ile Gln
195 200 205
Asn Ile Ser His Leu Ile Leu His Leu Lys Gln Pro Val Leu Leu Leu
210 215 220
Asp Ile Leu Ile Asp Ile Val Ser Ser Leu Asp Tyr Leu Glu Leu Arg
225 230 235 240
Asp Thr Asn Leu His Thr Phe Tyr Phe Ser Glu Ala Ser Ile Ser Glu
245 250 255
Val Asn Thr Ser Val Lys Lys Leu Ile Phe Arg Asn Val Gln Phe Thr
260 265 270
Asp Glu Ser Phe Val Glu Val Val Lys Leu Phe Asn Tyr Val Ser Gly
275 280 285
Ile Leu Glu Val Glu Phe Asp Asp Cys Thr His Asp Gly Ile Gly Asp
290 295 300
Phe Thr Ala Leu Thr Leu Asn Arg Ile Arg Tyr Leu Gly Asn Val Glu
305 310 315 320
Thr Leu Thr Ile Arg Lys Leu His Ile Pro Gln Phe Phe Leu Phe Tyr
325 330 335
Asp Leu Ser Ser Ile Tyr Pro Leu Thr Gly Lys Val Lys Arg Val Thr
340 345 350
Ile Glu Asn Ser Lys Val Phe Leu Val Pro Cys Leu Leu Ser Gln His
355 360 365
Leu Lys Ser Leu Glu Tyr Leu Asp Leu Ser Glu Asn Leu Met Ser Glu
370 375 380
Glu Thr Leu Arg Asn Ser Ala Cys Glu His Ala Trp Pro Phe Leu Gln
385 390 395 400
Thr Leu Val Leu Arg Gln Asn Arg Leu Lys Ser Leu Glu Lys Thr Gly
405 410 415
Glu Leu Leu Leu Thr Leu Lys Asn Leu Asn Asn Leu Asp Ile Ser Lys
420 425 430
Asn Asn Phe Leu Ser Met Pro Glu Thr Cys Gln Trp Pro Gly Lys Met
435 440 445
Lys Gln Leu Asn Leu Ser Ser Thr Arg Ile His Ser Leu Thr Gln Cys
450 455 460
Leu Pro Gln Thr Leu Glu Ile Leu Asp Val Ser Asn Asn Asn Leu Asp
465 470 475 480
Ser Phe Ser Leu Ile Leu Pro Gln Leu Lys Glu Leu Tyr Ile Ser Arg
485 490 495
Asn Lys Leu Lys Thr Leu Pro Asp Ala Ser Phe Leu Pro Val Leu Ser
500 505 510
Val Met Arg Ile Ser Gly Asn Ile Ile Asn Thr Phe Ser Lys Glu Gln
515 520 525
Leu Asp Ser Phe Pro Gln Leu Lys Ala Leu Glu Ala Gly Gly Asn Asn
530 535 540
Phe Ile Cys Ser Cys Asp Phe Leu Ser Phe Thr Gln Gly Gln Gln Ala
545 550 555 560
Leu Ala Arg Val Leu Val Asp Trp Pro Asp Gly Tyr Arg Cys Asp Ala
565 570 575
Pro Ser His Val Arg Gly Gln Arg Val Gln Asp Ala Arg Leu Ser Leu
580 585 590
Ser Glu Cys His Arg Ile Ile Ser Val Ser Val Val Thr Val Leu Leu
595 600 605
Val Ser Val Val Gly Val Leu Val Tyr Lys Phe Tyr Phe His Leu Met
610 615 620
Leu Leu Ala Gly Cys Lys Lys Tyr Gly Arg Gly Glu Ser Thr Tyr Asp
625 630 635 640
Ala Phe Val Ile Tyr Ser Ser Gln Asp Glu Ala Trp Val Arg Asn Glu
645 650 655
Leu Val Lys Asn Leu Glu Glu Gly Val Pro Pro Phe Gln Leu Cys Leu
660 665 670
His Tyr Arg Asp Phe Ile Pro Gly Val Ala Ile Ala Ala Asn Ile Ile
675 680 685
Gln Glu Gly Phe His Lys Ser Arg Lys Val Ile Val Val Val Ser Gln
690 695 700
His Phe Ile Gln Ser Arg Trp Cys Ile Phe Glu Tyr Glu Ile Ala Gln
705 710 715 720
Thr Trp Gln Phe Leu Ser Ser Arg Ala Gly Ile Ile Phe Ile Val Leu
725 730 735
Gln Lys Leu Glu Lys Ser Leu Leu Arg Gln Gln Val Glu Leu Tyr Arg
740 745 750
Leu Leu Asn Arg Asn Thr Tyr Leu Glu Trp Glu Asp Ser Val Leu Gly
755 760 765
Arg His Val Phe Trp Arg Arg Leu Arg Lys Ala Leu Leu Ala Gly Lys
770 775 780
Pro Arg Ser Pro Glu Gly Thr Ala Asp Ala Glu Thr Asn Pro Gln Glu
785 790 795 800
Ala Thr Thr Ser Thr
805
<210> 3
<211> 5599
<212> DNA
<213> 人工序列(Artificial sequence)
<400> 3
actacacatt atagatgact gggcccaagc tatggggtat ctactcaatt cagaatgaat 60
atacagcaga acatttttca taagactaac tgattaaaca aaatatgtac tgagaaatta 120
aagaaatgat aatctatcaa cctctcatga actaccctga atcttcacca caagagacat 180
cactgcgtca acagaaaggt taaaaatgcc atttcatcac atactttgga atttttggca 240
gcaaggaatg gtgtcaatca ttatttatca taggtacatt acactttgag gaatgactga 300
atttttttct ttttacaaat aaccgccagt tgtattacta aatatatact atctcataat 360
tttgcagggt tgaagtttca tttgtcctaa ataacaaaaa ataagaaaac tttattttat 420
tcataatcca ctttttcact ggtaagatgg atccaacaat cactgattgt ctttagcttc 480
tgcccaatat ttcataaatt aattgaatat gaagaggata acgtaaaagt ggttgtctgc 540
acatatgcat tttgaacaat attattttcc tgtccaaagg gaaaaaatga agtggccatc 600
ctcctccaga aaggccaaaa taagtcccca aagtatgtac atttctagac acaaaacact 660
atcagaagac agtaaaaatt tcaagcatgc tggtaaaata aattctatga aaaggccttc 720
tatgctctta aggaaatgat cttacgttaa gatctcaaag gagttccaaa caggaaatac 780
aagttaaaaa caattatttt ataaggtagc tttcatatta tataaaagac aacttacaga 840
catttgtatc tatgttaaac agccagtatg agtgacacca tacttatctg atatggcatc 900
ttttatatgt gatagtaata agaattagaa ccacctaaaa tgttatccat taaaggttct 960
gtttcattat tccccttgca acactgaaag acaatgcaag taacatttgt gaatgacaaa 1020
actcatttcc attctttctg gttcccgcca caaacgccag agtcatacta cacattatag 1080
atttaattaa gtcgactaga tgaaggagag cctttctctc tgggcaagag cggtgcaatg 1140
gtgtgtaaag gtagctgaga agacgaaaag ggcaagcatc ttcctgctac caggctgggg 1200
aggcccaggc ccacgacccc gaggagaggg aacgcaggga gactgaggtg acccttcttt 1260
cccccggggc ccggtcgtgt ggttcggtgt ctcttttctg ttggaccctt accttgaccc 1320
aggcgctgcc ggggcctggg cccgggctgc ggcgcacggc actcccggga ggcagcgaga 1380
ctcgagttag gcccaacgcg gcgccacggc gtttcctggc cgggaatggc ccgtacccgt 1440
gaggtggggg tggggggcag aaaaggcgga gcgagcccga ggcggggagg gggagggcca 1500
ggggcggagg gggccggcac tactgtgttg gcggactggc gggactaggg ctgcgtgagt 1560
ctctgagcgc aggcgggcgg cggccgcccc tcccccggcg gcggcagcgg cggcagcggc 1620
ggcagctcac tcagcccgct gcccgagcgg aaacgccact gaccgcacgg ggattcccag 1680
tgccggcgcc aggggcacgc gggacacgcc ccctcccgcc gcgccattgg cctctccgcc 1740
caccgcccca cacttattgg ccggtgcgcc gccaatcagc ggaggctgcc ggggccgcct 1800
aaagaagagg ctgtgctttg gggctccggc tcctcagaga gcctcggcta ggtaggggat 1860
cgggactctg gcgggagggc ggcttggtgc gtttgcgggg atccgaattc atgccacgtg 1920
ctttgtggac agcgtgggtc tgggctgtaa tcagcgtgtt cacggaagga gagcagaaac 1980
tcatctctga agaggatctg gcctctgatc aggcttcttc tctgtcttgt gacccaactg 2040
gtgtctgcga tggccattcc agatctttaa actccatccc ctctggtctc acagcaggtg 2100
tgaaaagcct tgacctgtcc gacaacgaga tcacctatgt cggcaacaga gacctgcaga 2160
ggtgtgtgaa cctgaagact ctgaggctgg gggccaatga aattcacacg gtggaggaag 2220
attctttttt tcacctgagg aatcttgaat atttggactt atcctataat cgcttatcta 2280
acttatcatc ttcctggttc aggtcccttt acgtcttgaa attcttaaac ttactgggaa 2340
atttatacaa aacacttggg gaaacatctc ttttttctca tctcccaaat ctgcggaccc 2400
tgaaagtagg aaatagtaac agcttcactc agattcatga aaaggatttc actggactga 2460
cttttcttga ggagcttgag atcagtgctc aaaatctgca gttatatgtg ccaaagagtt 2520
taaagtcgat ccagaacatt agccatctga ttcttcatct gaagcagcct gttttactcc 2580
tggacattct tatagatatt gtaagttcct tagattattt agaactgaga gatactaatt 2640
tgcacacttt ctatttttca gaagcatcca tcagtgaagt taatacatca gttaaaaagc 2700
ttatatttag aaatgtgcaa ttcaccgatg aaagttttgt tgaagttgtc aaactgttta 2760
actatgtttc tgggatcttg gaagtagagt ttgatgactg tacccatgat ggaattggcg 2820
attttacagc actgactttg aacagaatta gatacctagg taacgtggag acgttaacaa 2880
tacggaagtt gcatatccca cagtttttct tattttatga tctgagtagt atatatccac 2940
tcacaggcaa agttaaaaga gtcacaatag aaaacagtaa ggttttcctg gttccttgtt 3000
tactttcaca acatttaaaa tcactagaat atttggatct cagtgaaaac ttaatgtctg 3060
aagaaacctt gagaaactca gcctgtgagc atgcctggcc cttccttcaa accctggttt 3120
taaggcagaa tcgtttgaaa tcactagaaa aaactggaga acttttgctt actctgaaaa 3180
atctgaataa ccttgatatc agtaagaata attttctttc aatgcctgaa acttgtcagt 3240
ggccaggaaa aatgaaacag ttgaacttat ccagcacgag gatacacagt ttaacccagt 3300
gccttcccca gaccctggaa attttagatg ttagcaataa caatctcgat tcattttctt 3360
tgattttgcc gcaactcaaa gaactttata tttccagaaa taagttgaag actctaccag 3420
atgcctcctt cttacccgtg ttatcagtta tgagaattag cggaaatata ataaatactt 3480
tctcgaagga acaacttgat tcttttccac aactgaaggc tttggaggcc ggtggcaaca 3540
acttcatttg ctcctgtgac ttcctgtcct tcacacaggg acagcaggca ctggcccgtg 3600
tcctggtcga ctggccagat ggctaccgct gtgacgctcc ctcgcacgtg cggggccagc 3660
gggtgcagga cgcccggctc tccctttctg aatgccaccg gatcatcagc gtgtcggttg 3720
tcactgtact cctggtatct gtggtaggag tcctagtcta taagttctat ttccacctga 3780
tgcttcttgc tggctgcaaa aagtatggca gaggtgaaag cacctatgat gcctttgtaa 3840
tctactcgag ccaggatgaa gcctgggtgc ggaatgaact ggtaaagaac ttggaggagg 3900
gcgtgccccc ctttcagctc tgccttcact acagggactt tattcctggg gtggccatcg 3960
ccgccaacat catccaggaa ggtttccaca agagccgtaa ggtgattgtc gtggtgtccc 4020
agcacttcat ccagagccga tggtgtatct tcgagtatga gattgcccag acctggcagt 4080
ttctgagcag ccgtgctggc atcatcttca tcgtcctgca gaagctggag aagtctctcc 4140
tgcggcagca ggtggaactc tatcgccttc tgaacaggaa cacctacctg gagtgggagg 4200
acagtgtcct ggggcggcat gtcttctgga gaagactcag aaaagccttg ctggctggta 4260
aaccccggag tccagaagga acagcagatg cagagaccaa cccgcaggaa gcgaccacct 4320
ccacctgaca cgtgctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 4380
gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 4440
tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 4500
caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggt 4560
ctagagggtt catttacaca aaaccgaaca tactaaaatt ctgaaggaaa aaaaaaatat 4620
atatatatat aggatgcctg agtcttaggg ggagacaata acaggcatct atctgttaag 4680
tatcagatgt aactgatggt atattgggct ggtgattctc cttcaagtac ttccatattt 4740
aacaaactgt tctgcttaat cagagatctt cctaaactgt gggaaccgtg gaagaagaca 4800
gactacataa ggggcctaga tgctatcatc tcaagtatcc tgtatactag atttattcaa 4860
tttctgagta ttccttctag tctgtcatat ttcaaatgga tcactgtcaa acatatagat 4920
ccttgtgatc ttagacaact atactcaaca catattataa atcaatatca caggtcaaat 4980
tctaaagaaa taatctaatt tggagaaatc tattactaac acagaaaata agaaaatact 5040
tgatatttat acatagaaat cgttctgcaa aggacagttg tttctagagt ttaagaccct 5100
aaatatagta aaatgtttag caaaacatct gttcaataag gtaactatgg tctaacaact 5160
ggaaggaaga agacgtacaa gtattaaaat tgatttaatg attaaaaaga tggaggaaaa 5220
catgccagct gcagaaaaat caactgatat tttctccatt ctggttctca tgaagatcct 5280
taaattctta ttaccttggt tttaaaagga ttgaaaaatt aaaactcaaa tggaacttct 5340
ggtcctatcc aagcaaaatc cccagattaa aaccaaaaat ctgccacaaa aaccaaacag 5400
aaaagtcttt aactacggtt cagtttccaa atgctgatat ccataaatgg gtagaaggtt 5460
atgaattgct aattctcatt tccccaccaa taaaatgcaa accagcacag cagtgaaaat 5520
tcccaggaaa gcaaggtttc cgtcttagcc ctgaagcagc cattgcctaa gtgcagctcc 5580
ctgcagccaa cagcattaa 5599
Claims (9)
1.嵌合蛋白质,其特征在于,所述嵌合蛋白质由TLR2的胞外区和TLR4的跨膜区与胞内区组成。
2.根据权利要求1所述的嵌合蛋白质,其特征在于,所述嵌合蛋白质还包括信号肽和/或标签蛋白。
3.根据权利要求1或2所述的嵌合蛋白质,其特征在于,所述TLR2的胞外区为氨基酸序列是SEQ ID No.2的第31-597位的蛋白质;所述TLR4的跨膜区与胞内区为氨基酸序列是SEQID No.2的第598-805位的蛋白质。
4.根据权利要求1-3任一所述的嵌合蛋白质,其特征在于,所述嵌合蛋白质为下述任一种蛋白质:
A1)氨基酸序列是SEQ ID No.2的蛋白质;
A2)氨基酸序列是SEQ ID No.2的第31-805位的蛋白质。
5.生物材料,其特征在于,所述生物材料为下述任一种:
B1)编码权利要求1-4中任一所述的嵌合蛋白质的核酸分子;
B2)含有B1)所述核酸分子的表达盒;
B3)含有B1)所述核酸分子的重组载体、或含有B2)所述表达盒的重组载体;
B4)含有B1)所述核酸分子的重组微生物、或含有B2)所述表达盒的重组微生物、或含有B3)所述重组载体的重组微生物;
B5)含有B1)所述核酸分子的重组细胞、或含有B2)所述表达盒的重组细胞、或含有B3)所述重组载体的重组细胞。
6.根据权利要求5所述的生物材料,其特征在于,所述核酸分子为下述任一种:
C1)编码序列是SEQ ID No.1的DNA分子;
C2)编码序列是SEQ ID No.1的第91-2418位的DNA分子;
C3)与C1)或C2)限定的核苷酸序列具有75%或75%以上同一性,且编码权利要求3所述嵌合蛋白质的DNA分子。
7.应用,其特征在于,所述应用为M1或M2:
M1、权利要求5所述的生物材料在制备增强动物巨噬细胞对金黄色葡萄球菌的吞噬能力的产品中的应用,
M2、权利要求5所述的生物材料在制备增强动物细胞对金黄色葡萄球菌的抗性的产品中的应用。
8.根据权利要求7所述的应用,其特征在于,所述动物为山羊。
9.权利要求1-4中任一所述的嵌合蛋白质和/或权利要求5或6所述的生物材料在制备增强动物免疫力和/或提高动物抗病能力的产品中应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110834140.9A CN113501885B (zh) | 2021-07-20 | 2021-07-20 | 嵌合抗病基因及其相关生物材料与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110834140.9A CN113501885B (zh) | 2021-07-20 | 2021-07-20 | 嵌合抗病基因及其相关生物材料与应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113501885A CN113501885A (zh) | 2021-10-15 |
CN113501885B true CN113501885B (zh) | 2023-02-24 |
Family
ID=78014251
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110834140.9A Active CN113501885B (zh) | 2021-07-20 | 2021-07-20 | 嵌合抗病基因及其相关生物材料与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113501885B (zh) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017024440A1 (zh) * | 2015-08-07 | 2017-02-16 | 深圳市体内生物医药科技有限公司 | 含Toll样受体胞内结构域的嵌合抗原受体 |
CN108853144A (zh) * | 2017-05-16 | 2018-11-23 | 科济生物医药(上海)有限公司 | Toll样受体激动剂与免疫效应细胞的联用 |
CN109868277A (zh) * | 2019-03-13 | 2019-06-11 | 浙江海洋大学 | 曼氏无针乌贼Toll样受体基因及其免疫防御功能 |
-
2021
- 2021-07-20 CN CN202110834140.9A patent/CN113501885B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017024440A1 (zh) * | 2015-08-07 | 2017-02-16 | 深圳市体内生物医药科技有限公司 | 含Toll样受体胞内结构域的嵌合抗原受体 |
CN108853144A (zh) * | 2017-05-16 | 2018-11-23 | 科济生物医药(上海)有限公司 | Toll样受体激动剂与免疫效应细胞的联用 |
CN109868277A (zh) * | 2019-03-13 | 2019-06-11 | 浙江海洋大学 | 曼氏无针乌贼Toll样受体基因及其免疫防御功能 |
Non-Patent Citations (2)
Title |
---|
"Mastitis Increases Mammary mRNA Abundance of β-Defensin 5,Toll-Like-Receptor 2 (TLR2), and TLR4 but Not TLR9 in Cattle";T. Goldammer等;《MASTITIS AND INNATE IMMUNE DEFENSE》;20040131;第11卷(第1期);全文 * |
"Toll样受体研究进展";王德成等;《动物医学进展》;20080229;第29卷(第2期);全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN113501885A (zh) | 2021-10-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US10597648B2 (en) | Engineered cascade components and cascade complexes | |
CN111757889B (zh) | 新型CRISPR/Cas12f酶和系统 | |
Chen et al. | WSSV infection activates STAT in shrimp | |
AU2019381258A1 (en) | CRISPR-Cas12j enzyme and system | |
CN113881652B (zh) | 新型Cas酶和系统以及应用 | |
CN115851665A (zh) | 工程化的Cas12i核酸酶及其效应蛋白以及用途 | |
CN110643600A (zh) | 用于切割靶dna的系统及其用途 | |
CN113015798B (zh) | CRISPR-Cas12a酶和系统 | |
KR102508182B1 (ko) | 형질전환 대식세포, 키메라 항원 수용체, 및 관련 방법 | |
CN112105728A (zh) | CRISPR/Cas效应蛋白及系统 | |
CN112020560A (zh) | 一种RNA编辑的CRISPR/Cas效应蛋白及系统 | |
AU2018278516A1 (en) | Trait selection in avians | |
RU2746486C2 (ru) | Универсальный антителоопосредованный биосенсор | |
CN114134211B (zh) | Usp30基因作为靶点在抑制塞内卡谷病毒复制中的应用 | |
CN113501885B (zh) | 嵌合抗病基因及其相关生物材料与应用 | |
CN110577934B (zh) | 敲低tlr4基因猪肺泡巨噬细胞系的构建方法及其应用 | |
CN109021086B (zh) | 一种抗菌肽天蚕素a突变体及其编码基因、制备方法和应用 | |
CN101161676A (zh) | 抑制hiv复制的突变型apobec3g分子及其应用 | |
US20130023643A1 (en) | Nuclear localization signal peptides derived from vp2 protein of chicken anemia virus and uses of said peptides | |
CN113502299B (zh) | 一种抗金黄色葡萄球菌遗传修饰山羊生产方法 | |
WO2023138617A1 (zh) | 工程化的CasX核酸酶、效应蛋白及其用途 | |
WO2023234409A1 (ja) | ゲノム編集の効率を高めるための組成物及びその利用 | |
CN113372428B (zh) | 文昌鱼C型凝集素BjCTL4基因及其应用 | |
CN101603045B (zh) | 利用家畜乳腺高效生产人溶菌酶 | |
CN116769754A (zh) | 融合蛋白及其在基因编辑中的应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |