CN111549054A - CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用 - Google Patents
CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用 Download PDFInfo
- Publication number
- CN111549054A CN111549054A CN202010443155.8A CN202010443155A CN111549054A CN 111549054 A CN111549054 A CN 111549054A CN 202010443155 A CN202010443155 A CN 202010443155A CN 111549054 A CN111549054 A CN 111549054A
- Authority
- CN
- China
- Prior art keywords
- rfxcas13d
- crispr
- vector
- lys
- 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.)
- Pending
Links
- 239000013598 vector Substances 0.000 title claims abstract description 48
- 108091033409 CRISPR Proteins 0.000 title claims abstract description 25
- 238000010354 CRISPR gene editing Methods 0.000 title claims abstract description 25
- 241001493065 dsRNA viruses Species 0.000 title claims abstract description 25
- 108020005089 Plant RNA Proteins 0.000 title claims abstract description 21
- 238000010276 construction Methods 0.000 title claims abstract description 14
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 43
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims abstract description 15
- 241000723838 Turnip mosaic virus Species 0.000 claims abstract description 15
- 241000700605 Viruses Species 0.000 claims abstract description 12
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 11
- 208000015181 infectious disease Diseases 0.000 claims abstract description 9
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 claims abstract description 7
- 101710090029 Replication-associated protein A Proteins 0.000 claims abstract description 7
- 108700007698 Genetic Terminator Regions Proteins 0.000 claims abstract description 6
- 108091034057 RNA (poly(A)) Proteins 0.000 claims abstract description 6
- 230000010076 replication Effects 0.000 claims abstract description 6
- 230000002458 infectious effect Effects 0.000 claims abstract description 4
- 201000010099 disease Diseases 0.000 claims abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 3
- 239000012634 fragment Substances 0.000 claims description 24
- 239000013612 plasmid Substances 0.000 claims description 22
- 241000589158 Agrobacterium Species 0.000 claims description 10
- 102000012410 DNA Ligases Human genes 0.000 claims description 6
- 108010061982 DNA Ligases Proteins 0.000 claims description 6
- 241000207746 Nicotiana benthamiana Species 0.000 claims description 6
- 238000009825 accumulation Methods 0.000 claims description 5
- 238000003776 cleavage reaction Methods 0.000 claims description 4
- 238000010367 cloning Methods 0.000 claims description 4
- 230000029087 digestion Effects 0.000 claims description 4
- 238000000034 method Methods 0.000 claims description 4
- 230000007017 scission Effects 0.000 claims description 4
- 238000012163 sequencing technique Methods 0.000 claims description 4
- 230000002194 synthesizing effect Effects 0.000 claims description 4
- 238000012795 verification Methods 0.000 claims description 4
- 239000013603 viral vector Substances 0.000 claims description 4
- 125000006850 spacer group Chemical group 0.000 claims description 3
- 238000001976 enzyme digestion Methods 0.000 claims description 2
- 230000030279 gene silencing Effects 0.000 claims description 2
- 230000009368 gene silencing by RNA Effects 0.000 claims description 2
- 238000003860 storage Methods 0.000 claims description 2
- 208000024891 symptom Diseases 0.000 claims description 2
- 238000001262 western blot Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims 2
- 238000005070 sampling Methods 0.000 claims 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 230000001131 transforming effect Effects 0.000 claims 1
- 241000196324 Embryophyta Species 0.000 abstract description 18
- 108091027544 Subgenomic mRNA Proteins 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 239000007850 fluorescent dye Substances 0.000 abstract 1
- 239000005090 green fluorescent protein Substances 0.000 description 15
- 108020004414 DNA Proteins 0.000 description 6
- 238000010362 genome editing Methods 0.000 description 6
- 108090000565 Capsid Proteins Proteins 0.000 description 4
- 102100023321 Ceruloplasmin Human genes 0.000 description 4
- 108010042407 Endonucleases Proteins 0.000 description 4
- 102000004533 Endonucleases Human genes 0.000 description 4
- 108010005233 alanylglutamic acid Proteins 0.000 description 4
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 4
- 108010038320 lysylphenylalanine Proteins 0.000 description 4
- YGKVNUAKYPGORG-AVGNSLFASA-N Tyr-Asp-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O YGKVNUAKYPGORG-AVGNSLFASA-N 0.000 description 3
- 238000012271 agricultural production Methods 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
- 238000005516 engineering process Methods 0.000 description 3
- 108010050848 glycylleucine Proteins 0.000 description 3
- 108010054155 lysyllysine Proteins 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- PAPSMOYMQDWIOR-AVGNSLFASA-N Arg-Lys-Val Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O PAPSMOYMQDWIOR-AVGNSLFASA-N 0.000 description 2
- UVTGNSWSRSCPLP-UHFFFAOYSA-N Arg-Tyr Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccc(O)cc1)C(=O)O UVTGNSWSRSCPLP-UHFFFAOYSA-N 0.000 description 2
- SLKLLQWZQHXYSV-CIUDSAMLSA-N Asn-Ala-Lys Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O SLKLLQWZQHXYSV-CIUDSAMLSA-N 0.000 description 2
- 108010077805 Bacterial Proteins Proteins 0.000 description 2
- OCJRHJZKGGSPRW-IUCAKERBSA-N Glu-Lys-Gly Chemical compound NCCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CCC(O)=O OCJRHJZKGGSPRW-IUCAKERBSA-N 0.000 description 2
- YKBUCXNNBYZYAY-MNXVOIDGSA-N Glu-Lys-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YKBUCXNNBYZYAY-MNXVOIDGSA-N 0.000 description 2
- OCDLPQDYTJPWNG-YUMQZZPRSA-N Gly-Asn-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)CN OCDLPQDYTJPWNG-YUMQZZPRSA-N 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- UZZXGLOJRZKYEL-DJFWLOJKSA-N His-Asn-Ile Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UZZXGLOJRZKYEL-DJFWLOJKSA-N 0.000 description 2
- 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 2
- JYXBNQOKPRQNQS-YTFOTSKYSA-N Lys-Ile-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JYXBNQOKPRQNQS-YTFOTSKYSA-N 0.000 description 2
- SKRGVGLIRUGANF-AVGNSLFASA-N Lys-Leu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SKRGVGLIRUGANF-AVGNSLFASA-N 0.000 description 2
- WBSCNDJQPKSPII-KKUMJFAQSA-N Lys-Lys-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O WBSCNDJQPKSPII-KKUMJFAQSA-N 0.000 description 2
- 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 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 2
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 2
- WIVCOAKLPICYGY-KKUMJFAQSA-N Phe-Asp-Lys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N WIVCOAKLPICYGY-KKUMJFAQSA-N 0.000 description 2
- OQTDZEJJWWAGJT-KKUMJFAQSA-N Phe-Lys-Asp Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O OQTDZEJJWWAGJT-KKUMJFAQSA-N 0.000 description 2
- 238000010459 TALEN Methods 0.000 description 2
- 108010043645 Transcription Activator-Like Effector Nucleases Proteins 0.000 description 2
- OJOBTAOGJIWAGB-UHFFFAOYSA-N acetosyringone Chemical compound COC1=CC(C(C)=O)=CC(OC)=C1O OJOBTAOGJIWAGB-UHFFFAOYSA-N 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 2
- 108010049041 glutamylalanine Proteins 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 108010057821 leucylproline Proteins 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 108010003700 lysyl aspartic acid Proteins 0.000 description 2
- 230000006780 non-homologous end joining Effects 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 230000003007 single stranded DNA break Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- YYYARFHFWYKNLF-UHFFFAOYSA-N 4-[(2,4-dimethylphenyl)diazenyl]-3-hydroxynaphthalene-2,7-disulfonic acid Chemical compound CC1=CC(C)=CC=C1N=NC1=C(O)C(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=C12 YYYARFHFWYKNLF-UHFFFAOYSA-N 0.000 description 1
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 1
- AAQGRPOPTAUUBM-ZLUOBGJFSA-N Ala-Ala-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O AAQGRPOPTAUUBM-ZLUOBGJFSA-N 0.000 description 1
- FJVAQLJNTSUQPY-CIUDSAMLSA-N Ala-Ala-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN FJVAQLJNTSUQPY-CIUDSAMLSA-N 0.000 description 1
- UGLPMYSCWHTZQU-AUTRQRHGSA-N Ala-Ala-Tyr Chemical compound C[C@H]([NH3+])C(=O)N[C@@H](C)C(=O)N[C@H](C([O-])=O)CC1=CC=C(O)C=C1 UGLPMYSCWHTZQU-AUTRQRHGSA-N 0.000 description 1
- XEXJJJRVTFGWIC-FXQIFTODSA-N Ala-Asn-Arg Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N XEXJJJRVTFGWIC-FXQIFTODSA-N 0.000 description 1
- XCVRVWZTXPCYJT-BIIVOSGPSA-N Ala-Asn-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N XCVRVWZTXPCYJT-BIIVOSGPSA-N 0.000 description 1
- XQJAFSDFQZPYCU-UWJYBYFXSA-N Ala-Asn-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N XQJAFSDFQZPYCU-UWJYBYFXSA-N 0.000 description 1
- 108010040956 Ala-Asp-Glu-Leu Proteins 0.000 description 1
- LSLIRHLIUDVNBN-CIUDSAMLSA-N Ala-Asp-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN LSLIRHLIUDVNBN-CIUDSAMLSA-N 0.000 description 1
- BTYTYHBSJKQBQA-GCJQMDKQSA-N Ala-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C)N)O BTYTYHBSJKQBQA-GCJQMDKQSA-N 0.000 description 1
- ZDYNWWQXFRUOEO-XDTLVQLUSA-N Ala-Gln-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZDYNWWQXFRUOEO-XDTLVQLUSA-N 0.000 description 1
- NWVVKQZOVSTDBQ-CIUDSAMLSA-N Ala-Glu-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NWVVKQZOVSTDBQ-CIUDSAMLSA-N 0.000 description 1
- BGNLUHXLSAQYRQ-FXQIFTODSA-N Ala-Glu-Gln Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O BGNLUHXLSAQYRQ-FXQIFTODSA-N 0.000 description 1
- UHMQKOBNPRAZGB-CIUDSAMLSA-N Ala-Glu-Met Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCSC)C(=O)O)N UHMQKOBNPRAZGB-CIUDSAMLSA-N 0.000 description 1
- ROLXPVQSRCPVGK-XDTLVQLUSA-N Ala-Glu-Tyr Chemical compound N[C@@H](C)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O ROLXPVQSRCPVGK-XDTLVQLUSA-N 0.000 description 1
- RZZMZYZXNJRPOJ-BJDJZHNGSA-N Ala-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](C)N RZZMZYZXNJRPOJ-BJDJZHNGSA-N 0.000 description 1
- HHRAXZAYZFFRAM-CIUDSAMLSA-N Ala-Leu-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O HHRAXZAYZFFRAM-CIUDSAMLSA-N 0.000 description 1
- VHVVPYOJIIQCKS-QEJZJMRPSA-N Ala-Leu-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 VHVVPYOJIIQCKS-QEJZJMRPSA-N 0.000 description 1
- MEFILNJXAVSUTO-JXUBOQSCSA-N Ala-Leu-Thr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MEFILNJXAVSUTO-JXUBOQSCSA-N 0.000 description 1
- RGQCNKIDEQJEBT-CQDKDKBSSA-N Ala-Leu-Tyr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 RGQCNKIDEQJEBT-CQDKDKBSSA-N 0.000 description 1
- MFMDKJIPHSWSBM-GUBZILKMSA-N Ala-Lys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O MFMDKJIPHSWSBM-GUBZILKMSA-N 0.000 description 1
- SUHLZMHFRALVSY-YUMQZZPRSA-N Ala-Lys-Gly Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)NCC(O)=O SUHLZMHFRALVSY-YUMQZZPRSA-N 0.000 description 1
- OINVDEKBKBCPLX-JXUBOQSCSA-N Ala-Lys-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OINVDEKBKBCPLX-JXUBOQSCSA-N 0.000 description 1
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 1
- CJQAEJMHBAOQHA-DLOVCJGASA-N Ala-Phe-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)N)C(=O)O)N CJQAEJMHBAOQHA-DLOVCJGASA-N 0.000 description 1
- YCRAFFCYWOUEOF-DLOVCJGASA-N Ala-Phe-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 YCRAFFCYWOUEOF-DLOVCJGASA-N 0.000 description 1
- ZJLORAAXDAJLDC-CQDKDKBSSA-N Ala-Tyr-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(O)=O ZJLORAAXDAJLDC-CQDKDKBSSA-N 0.000 description 1
- XSLGWYYNOSUMRM-ZKWXMUAHSA-N Ala-Val-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O XSLGWYYNOSUMRM-ZKWXMUAHSA-N 0.000 description 1
- RVDVDRUZWZIBJQ-CIUDSAMLSA-N Arg-Asn-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O RVDVDRUZWZIBJQ-CIUDSAMLSA-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
- RWCLSUOSKWTXLA-FXQIFTODSA-N Arg-Asp-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O RWCLSUOSKWTXLA-FXQIFTODSA-N 0.000 description 1
- JCAISGGAOQXEHJ-ZPFDUUQYSA-N Arg-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCN=C(N)N)N JCAISGGAOQXEHJ-ZPFDUUQYSA-N 0.000 description 1
- OGUPCHKBOKJFMA-SRVKXCTJSA-N Arg-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N OGUPCHKBOKJFMA-SRVKXCTJSA-N 0.000 description 1
- UBCPNBUIQNMDNH-NAKRPEOUSA-N Arg-Ile-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O UBCPNBUIQNMDNH-NAKRPEOUSA-N 0.000 description 1
- HJDNZFIYILEIKR-OSUNSFLBSA-N Arg-Ile-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HJDNZFIYILEIKR-OSUNSFLBSA-N 0.000 description 1
- YBZMTKUDWXZLIX-UWVGGRQHSA-N Arg-Leu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YBZMTKUDWXZLIX-UWVGGRQHSA-N 0.000 description 1
- FIQKRDXFTANIEJ-ULQDDVLXSA-N Arg-Phe-His Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N FIQKRDXFTANIEJ-ULQDDVLXSA-N 0.000 description 1
- IGFJVXOATGZTHD-UHFFFAOYSA-N Arg-Phe-His Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccccc1)C(=O)NC(Cc2c[nH]cn2)C(=O)O IGFJVXOATGZTHD-UHFFFAOYSA-N 0.000 description 1
- OQPAZKMGCWPERI-GUBZILKMSA-N Arg-Ser-Val Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O OQPAZKMGCWPERI-GUBZILKMSA-N 0.000 description 1
- VLIJAPRTSXSGFY-STQMWFEESA-N Arg-Tyr-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=C(O)C=C1 VLIJAPRTSXSGFY-STQMWFEESA-N 0.000 description 1
- HZPSDHRYYIORKR-WHFBIAKZSA-N Asn-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CC(N)=O HZPSDHRYYIORKR-WHFBIAKZSA-N 0.000 description 1
- QHBMKQWOIYJYMI-BYULHYEWSA-N Asn-Asn-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O QHBMKQWOIYJYMI-BYULHYEWSA-N 0.000 description 1
- WVCJSDCHTUTONA-FXQIFTODSA-N Asn-Asp-Arg Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WVCJSDCHTUTONA-FXQIFTODSA-N 0.000 description 1
- UGXVKHRDGLYFKR-CIUDSAMLSA-N Asn-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(N)=O UGXVKHRDGLYFKR-CIUDSAMLSA-N 0.000 description 1
- QISZHYWZHJRDAO-CIUDSAMLSA-N Asn-Asp-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N QISZHYWZHJRDAO-CIUDSAMLSA-N 0.000 description 1
- ULRPXVNMIIYDDJ-ACZMJKKPSA-N Asn-Glu-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC(=O)N)N ULRPXVNMIIYDDJ-ACZMJKKPSA-N 0.000 description 1
- MECFLTFREHAZLH-ACZMJKKPSA-N Asn-Glu-Cys Chemical compound C(CC(=O)O)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(=O)N)N MECFLTFREHAZLH-ACZMJKKPSA-N 0.000 description 1
- BZMWJLLUAKSIMH-FXQIFTODSA-N Asn-Glu-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O BZMWJLLUAKSIMH-FXQIFTODSA-N 0.000 description 1
- GNKVBRYFXYWXAB-WDSKDSINSA-N Asn-Glu-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O GNKVBRYFXYWXAB-WDSKDSINSA-N 0.000 description 1
- JREOBWLIZLXRIS-GUBZILKMSA-N Asn-Glu-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O JREOBWLIZLXRIS-GUBZILKMSA-N 0.000 description 1
- QUAWOKPCAKCHQL-SRVKXCTJSA-N Asn-His-Lys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N QUAWOKPCAKCHQL-SRVKXCTJSA-N 0.000 description 1
- LVHMEJJWEXBMKK-GMOBBJLQSA-N Asn-Ile-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC(=O)N)N LVHMEJJWEXBMKK-GMOBBJLQSA-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
- 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
- HZZIFFOVHLWGCS-KKUMJFAQSA-N Asn-Phe-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O HZZIFFOVHLWGCS-KKUMJFAQSA-N 0.000 description 1
- XTMZYFMTYJNABC-ZLUOBGJFSA-N Asn-Ser-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)N XTMZYFMTYJNABC-ZLUOBGJFSA-N 0.000 description 1
- HPBNLFLSSQDFQW-WHFBIAKZSA-N Asn-Ser-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O HPBNLFLSSQDFQW-WHFBIAKZSA-N 0.000 description 1
- YSYTWUMRHSFODC-QWRGUYRKSA-N Asn-Tyr-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O YSYTWUMRHSFODC-QWRGUYRKSA-N 0.000 description 1
- MJIJBEYEHBKTIM-BYULHYEWSA-N Asn-Val-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N MJIJBEYEHBKTIM-BYULHYEWSA-N 0.000 description 1
- UWMIZBCTVWVMFI-FXQIFTODSA-N Asp-Ala-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O UWMIZBCTVWVMFI-FXQIFTODSA-N 0.000 description 1
- SDHFVYLZFBDSQT-DCAQKATOSA-N Asp-Arg-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(=O)O)N SDHFVYLZFBDSQT-DCAQKATOSA-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
- JDHOJQJMWBKHDB-CIUDSAMLSA-N Asp-Asn-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)O)N JDHOJQJMWBKHDB-CIUDSAMLSA-N 0.000 description 1
- UGIBTKGQVWFTGX-BIIVOSGPSA-N Asp-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)O)N)C(=O)O UGIBTKGQVWFTGX-BIIVOSGPSA-N 0.000 description 1
- SNAWMGHSCHKSDK-GUBZILKMSA-N Asp-Gln-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC(=O)O)N SNAWMGHSCHKSDK-GUBZILKMSA-N 0.000 description 1
- XJQRWGXKUSDEFI-ACZMJKKPSA-N Asp-Glu-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O XJQRWGXKUSDEFI-ACZMJKKPSA-N 0.000 description 1
- KHBLRHKVXICFMY-GUBZILKMSA-N Asp-Glu-Lys Chemical compound N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O KHBLRHKVXICFMY-GUBZILKMSA-N 0.000 description 1
- OVPHVTCDVYYTHN-AVGNSLFASA-N Asp-Glu-Phe Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 OVPHVTCDVYYTHN-AVGNSLFASA-N 0.000 description 1
- KTTCQQNRRLCIBC-GHCJXIJMSA-N Asp-Ile-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O KTTCQQNRRLCIBC-GHCJXIJMSA-N 0.000 description 1
- QNFRBNZGVVKBNJ-PEFMBERDSA-N Asp-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)O)N QNFRBNZGVVKBNJ-PEFMBERDSA-N 0.000 description 1
- NHSDEZURHWEZPN-SXTJYALSSA-N Asp-Ile-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)NC(=O)[C@H](CC(=O)O)N NHSDEZURHWEZPN-SXTJYALSSA-N 0.000 description 1
- PYXXJFRXIYAESU-PCBIJLKTSA-N Asp-Ile-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PYXXJFRXIYAESU-PCBIJLKTSA-N 0.000 description 1
- HXVILZUZXFLVEN-DCAQKATOSA-N Asp-Met-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O HXVILZUZXFLVEN-DCAQKATOSA-N 0.000 description 1
- GPPIDDWYKJPRES-YDHLFZDLSA-N Asp-Phe-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O GPPIDDWYKJPRES-YDHLFZDLSA-N 0.000 description 1
- KPSHWSWFPUDEGF-FXQIFTODSA-N Asp-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC(O)=O KPSHWSWFPUDEGF-FXQIFTODSA-N 0.000 description 1
- DRCOAZZDQRCGGP-GHCJXIJMSA-N Asp-Ser-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O DRCOAZZDQRCGGP-GHCJXIJMSA-N 0.000 description 1
- JJQGZGOEDSSHTE-FOHZUACHSA-N Asp-Thr-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O JJQGZGOEDSSHTE-FOHZUACHSA-N 0.000 description 1
- XMKXONRMGJXCJV-LAEOZQHASA-N Asp-Val-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O XMKXONRMGJXCJV-LAEOZQHASA-N 0.000 description 1
- 238000010453 CRISPR/Cas method Methods 0.000 description 1
- AEJSNWMRPXAKCW-WHFBIAKZSA-N Cys-Ala-Gly Chemical compound SC[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O AEJSNWMRPXAKCW-WHFBIAKZSA-N 0.000 description 1
- SAEVTQWAYDPXMU-KATARQTJSA-N Cys-Thr-Leu Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O SAEVTQWAYDPXMU-KATARQTJSA-N 0.000 description 1
- VIOQRFNAZDMVLO-NRPADANISA-N Cys-Val-Glu Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O VIOQRFNAZDMVLO-NRPADANISA-N 0.000 description 1
- 241000450599 DNA viruses Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- NUMFTVCBONFQIQ-DRZSPHRISA-N Gln-Ala-Phe Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O NUMFTVCBONFQIQ-DRZSPHRISA-N 0.000 description 1
- MWLYSLMKFXWZPW-ZPFDUUQYSA-N Gln-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CCC(N)=O MWLYSLMKFXWZPW-ZPFDUUQYSA-N 0.000 description 1
- HSHCEAUPUPJPTE-JYJNAYRXSA-N Gln-Leu-Tyr Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N HSHCEAUPUPJPTE-JYJNAYRXSA-N 0.000 description 1
- JNENSVNAUWONEZ-GUBZILKMSA-N Gln-Lys-Asn Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O JNENSVNAUWONEZ-GUBZILKMSA-N 0.000 description 1
- SDSMVVSHLAAOJL-UKJIMTQDSA-N Gln-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCC(=O)N)N SDSMVVSHLAAOJL-UKJIMTQDSA-N 0.000 description 1
- MXOODARRORARSU-ACZMJKKPSA-N Glu-Ala-Ser Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)O)N MXOODARRORARSU-ACZMJKKPSA-N 0.000 description 1
- CGYDXNKRIMJMLV-GUBZILKMSA-N Glu-Arg-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O CGYDXNKRIMJMLV-GUBZILKMSA-N 0.000 description 1
- GCYFUZJHAXJKKE-KKUMJFAQSA-N Glu-Arg-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O GCYFUZJHAXJKKE-KKUMJFAQSA-N 0.000 description 1
- AKJRHDMTEJXTPV-ACZMJKKPSA-N Glu-Asn-Ala Chemical compound C[C@H](NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCC(O)=O)C(O)=O AKJRHDMTEJXTPV-ACZMJKKPSA-N 0.000 description 1
- RDPOETHPAQEGDP-ACZMJKKPSA-N Glu-Asp-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O RDPOETHPAQEGDP-ACZMJKKPSA-N 0.000 description 1
- JVSBYEDSSRZQGV-GUBZILKMSA-N Glu-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCC(O)=O JVSBYEDSSRZQGV-GUBZILKMSA-N 0.000 description 1
- RAUDKMVXNOWDLS-WDSKDSINSA-N Glu-Gly-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O RAUDKMVXNOWDLS-WDSKDSINSA-N 0.000 description 1
- VSRCAOIHMGCIJK-SRVKXCTJSA-N Glu-Leu-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O VSRCAOIHMGCIJK-SRVKXCTJSA-N 0.000 description 1
- HRBYTAIBKPNZKQ-AVGNSLFASA-N Glu-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCC(O)=O HRBYTAIBKPNZKQ-AVGNSLFASA-N 0.000 description 1
- ITVBKCZZLJUUHI-HTUGSXCWSA-N Glu-Phe-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ITVBKCZZLJUUHI-HTUGSXCWSA-N 0.000 description 1
- WIKMTDVSCUJIPJ-CIUDSAMLSA-N Glu-Ser-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N WIKMTDVSCUJIPJ-CIUDSAMLSA-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
- HVKAAUOFFTUSAA-XDTLVQLUSA-N Glu-Tyr-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O HVKAAUOFFTUSAA-XDTLVQLUSA-N 0.000 description 1
- UZWUBBRJWFTHTD-LAEOZQHASA-N Glu-Val-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCC(O)=O UZWUBBRJWFTHTD-LAEOZQHASA-N 0.000 description 1
- XUDLUKYPXQDCRX-BQBZGAKWSA-N Gly-Arg-Asn Chemical compound [H]NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O XUDLUKYPXQDCRX-BQBZGAKWSA-N 0.000 description 1
- AIJAPFVDBFYNKN-WHFBIAKZSA-N Gly-Asn-Asp Chemical compound C([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)CN)C(=O)N AIJAPFVDBFYNKN-WHFBIAKZSA-N 0.000 description 1
- BULIVUZUDBHKKZ-WDSKDSINSA-N Gly-Gln-Asn Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O BULIVUZUDBHKKZ-WDSKDSINSA-N 0.000 description 1
- DGKBSGNCMCLDSL-BYULHYEWSA-N Gly-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)CN DGKBSGNCMCLDSL-BYULHYEWSA-N 0.000 description 1
- LLZXNUUIBOALNY-QWRGUYRKSA-N Gly-Leu-Lys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN LLZXNUUIBOALNY-QWRGUYRKSA-N 0.000 description 1
- BXICSAQLIHFDDL-YUMQZZPRSA-N Gly-Lys-Asn Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O BXICSAQLIHFDDL-YUMQZZPRSA-N 0.000 description 1
- LOEANKRDMMVOGZ-YUMQZZPRSA-N Gly-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)N[C@@H](CC(O)=O)C(O)=O LOEANKRDMMVOGZ-YUMQZZPRSA-N 0.000 description 1
- OQQKUTVULYLCDG-ONGXEEELSA-N Gly-Lys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)CN)C(O)=O OQQKUTVULYLCDG-ONGXEEELSA-N 0.000 description 1
- MTBIKIMYHUWBRX-QWRGUYRKSA-N Gly-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)CN MTBIKIMYHUWBRX-QWRGUYRKSA-N 0.000 description 1
- IGOYNRWLWHWAQO-JTQLQIEISA-N Gly-Phe-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 IGOYNRWLWHWAQO-JTQLQIEISA-N 0.000 description 1
- DHNXGWVNLFPOMQ-KBPBESRZSA-N Gly-Phe-His Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)CN DHNXGWVNLFPOMQ-KBPBESRZSA-N 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- TVQGUFGDVODUIF-LSJOCFKGSA-N His-Arg-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC1=CN=CN1)N TVQGUFGDVODUIF-LSJOCFKGSA-N 0.000 description 1
- OQDLKDUVMTUPPG-AVGNSLFASA-N His-Leu-Glu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O OQDLKDUVMTUPPG-AVGNSLFASA-N 0.000 description 1
- VSZALHITQINTGC-GHCJXIJMSA-N Ile-Ala-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CC(=O)O)C(=O)O)N VSZALHITQINTGC-GHCJXIJMSA-N 0.000 description 1
- RWIKBYVJQAJYDP-BJDJZHNGSA-N Ile-Ala-Lys Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN RWIKBYVJQAJYDP-BJDJZHNGSA-N 0.000 description 1
- ATXGFMOBVKSOMK-PEDHHIEDSA-N Ile-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N ATXGFMOBVKSOMK-PEDHHIEDSA-N 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
- QADCTXFNLZBZAB-GHCJXIJMSA-N Ile-Asn-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C)C(=O)O)N QADCTXFNLZBZAB-GHCJXIJMSA-N 0.000 description 1
- FJWYJQRCVNGEAQ-ZPFDUUQYSA-N Ile-Asn-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N FJWYJQRCVNGEAQ-ZPFDUUQYSA-N 0.000 description 1
- RPZFUIQVAPZLRH-GHCJXIJMSA-N Ile-Asp-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C)C(=O)O)N RPZFUIQVAPZLRH-GHCJXIJMSA-N 0.000 description 1
- UMYZBHKAVTXWIW-GMOBBJLQSA-N Ile-Asp-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N UMYZBHKAVTXWIW-GMOBBJLQSA-N 0.000 description 1
- IDAHFEPYTJJZFD-PEFMBERDSA-N Ile-Asp-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N IDAHFEPYTJJZFD-PEFMBERDSA-N 0.000 description 1
- DCQMJRSOGCYKTR-GHCJXIJMSA-N Ile-Asp-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O DCQMJRSOGCYKTR-GHCJXIJMSA-N 0.000 description 1
- WTOAPTKSZJJWKK-HTFCKZLJSA-N Ile-Cys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N WTOAPTKSZJJWKK-HTFCKZLJSA-N 0.000 description 1
- YBJWJQQBWRARLT-KBIXCLLPSA-N Ile-Gln-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O YBJWJQQBWRARLT-KBIXCLLPSA-N 0.000 description 1
- DFJJAVZIHDFOGQ-MNXVOIDGSA-N Ile-Glu-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N DFJJAVZIHDFOGQ-MNXVOIDGSA-N 0.000 description 1
- VOBYAKCXGQQFLR-LSJOCFKGSA-N Ile-Gly-Val Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O VOBYAKCXGQQFLR-LSJOCFKGSA-N 0.000 description 1
- KYLIZSDYWQQTFM-PEDHHIEDSA-N Ile-Ile-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CCCN=C(N)N KYLIZSDYWQQTFM-PEDHHIEDSA-N 0.000 description 1
- RVNOXPZHMUWCLW-GMOBBJLQSA-N Ile-Met-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(=O)N)C(=O)O)N RVNOXPZHMUWCLW-GMOBBJLQSA-N 0.000 description 1
- JHNJNTMTZHEDLJ-NAKRPEOUSA-N Ile-Ser-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O JHNJNTMTZHEDLJ-NAKRPEOUSA-N 0.000 description 1
- SAEWJTCJQVZQNZ-IUKAMOBKSA-N Ile-Thr-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N SAEWJTCJQVZQNZ-IUKAMOBKSA-N 0.000 description 1
- QGXQHJQPAPMACW-PPCPHDFISA-N Ile-Thr-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)O)N QGXQHJQPAPMACW-PPCPHDFISA-N 0.000 description 1
- BLFXHAFTNYZEQE-VKOGCVSHSA-N Ile-Trp-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N BLFXHAFTNYZEQE-VKOGCVSHSA-N 0.000 description 1
- HQLSBZFLOUHQJK-STECZYCISA-N Ile-Tyr-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N HQLSBZFLOUHQJK-STECZYCISA-N 0.000 description 1
- ZYVTXBXHIKGZMD-QSFUFRPTSA-N Ile-Val-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(=O)N)C(=O)O)N ZYVTXBXHIKGZMD-QSFUFRPTSA-N 0.000 description 1
- 206010061217 Infestation Diseases 0.000 description 1
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- HASRFYOMVPJRPU-SRVKXCTJSA-N Leu-Arg-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O HASRFYOMVPJRPU-SRVKXCTJSA-N 0.000 description 1
- KSZCCRIGNVSHFH-UWVGGRQHSA-N Leu-Arg-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O KSZCCRIGNVSHFH-UWVGGRQHSA-N 0.000 description 1
- UILIPCLTHRPCRB-XUXIUFHCSA-N Leu-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(C)C)N UILIPCLTHRPCRB-XUXIUFHCSA-N 0.000 description 1
- ULXYQAJWJGLCNR-YUMQZZPRSA-N Leu-Asp-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O ULXYQAJWJGLCNR-YUMQZZPRSA-N 0.000 description 1
- KTFHTMHHKXUYPW-ZPFDUUQYSA-N Leu-Asp-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KTFHTMHHKXUYPW-ZPFDUUQYSA-N 0.000 description 1
- MYGQXVYRZMKRDB-SRVKXCTJSA-N Leu-Asp-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN MYGQXVYRZMKRDB-SRVKXCTJSA-N 0.000 description 1
- WQWSMEOYXJTFRU-GUBZILKMSA-N Leu-Glu-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O WQWSMEOYXJTFRU-GUBZILKMSA-N 0.000 description 1
- HYMLKESRWLZDBR-WEDXCCLWSA-N Leu-Gly-Thr Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HYMLKESRWLZDBR-WEDXCCLWSA-N 0.000 description 1
- KXODZBLFVFSLAI-AVGNSLFASA-N Leu-His-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(C)C)CC1=CN=CN1 KXODZBLFVFSLAI-AVGNSLFASA-N 0.000 description 1
- USLNHQZCDQJBOV-ZPFDUUQYSA-N Leu-Ile-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O USLNHQZCDQJBOV-ZPFDUUQYSA-N 0.000 description 1
- HNDWYLYAYNBWMP-AJNGGQMLSA-N Leu-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(C)C)N HNDWYLYAYNBWMP-AJNGGQMLSA-N 0.000 description 1
- IFMPDNRWZZEZSL-SRVKXCTJSA-N Leu-Leu-Cys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(O)=O IFMPDNRWZZEZSL-SRVKXCTJSA-N 0.000 description 1
- LXKNSJLSGPNHSK-KKUMJFAQSA-N Leu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N LXKNSJLSGPNHSK-KKUMJFAQSA-N 0.000 description 1
- WXUOJXIGOPMDJM-SRVKXCTJSA-N Leu-Lys-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O WXUOJXIGOPMDJM-SRVKXCTJSA-N 0.000 description 1
- OZTZJMUZVAVJGY-BZSNNMDCSA-N Leu-Tyr-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N OZTZJMUZVAVJGY-BZSNNMDCSA-N 0.000 description 1
- VKVDRTGWLVZJOM-DCAQKATOSA-N Leu-Val-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O VKVDRTGWLVZJOM-DCAQKATOSA-N 0.000 description 1
- PNPYKQFJGRFYJE-GUBZILKMSA-N Lys-Ala-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O PNPYKQFJGRFYJE-GUBZILKMSA-N 0.000 description 1
- KCXUCYYZNZFGLL-SRVKXCTJSA-N Lys-Ala-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O KCXUCYYZNZFGLL-SRVKXCTJSA-N 0.000 description 1
- IRNSXVOWSXSULE-DCAQKATOSA-N Lys-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN IRNSXVOWSXSULE-DCAQKATOSA-N 0.000 description 1
- SJNZALDHDUYDBU-IHRRRGAJSA-N Lys-Arg-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(O)=O SJNZALDHDUYDBU-IHRRRGAJSA-N 0.000 description 1
- NQCJGQHHYZNUDK-DCAQKATOSA-N Lys-Arg-Ser Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CO)C(O)=O)CCCN=C(N)N NQCJGQHHYZNUDK-DCAQKATOSA-N 0.000 description 1
- GGAPIOORBXHMNY-ULQDDVLXSA-N Lys-Arg-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCCN)N)O GGAPIOORBXHMNY-ULQDDVLXSA-N 0.000 description 1
- HKCCVDWHHTVVPN-CIUDSAMLSA-N Lys-Asp-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O HKCCVDWHHTVVPN-CIUDSAMLSA-N 0.000 description 1
- LPAJOCKCPRZEAG-MNXVOIDGSA-N Lys-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CCCCN LPAJOCKCPRZEAG-MNXVOIDGSA-N 0.000 description 1
- VQXAVLQBQJMENB-SRVKXCTJSA-N Lys-Glu-Met Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O VQXAVLQBQJMENB-SRVKXCTJSA-N 0.000 description 1
- DUTMKEAPLLUGNO-JYJNAYRXSA-N Lys-Glu-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O DUTMKEAPLLUGNO-JYJNAYRXSA-N 0.000 description 1
- NKKFVJRLCCUJNA-QWRGUYRKSA-N Lys-Gly-Lys Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN NKKFVJRLCCUJNA-QWRGUYRKSA-N 0.000 description 1
- VLMNBMFYRMGEMB-QWRGUYRKSA-N Lys-His-Gly Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CNC=N1 VLMNBMFYRMGEMB-QWRGUYRKSA-N 0.000 description 1
- VUTWYNQUSJWBHO-BZSNNMDCSA-N Lys-Leu-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O VUTWYNQUSJWBHO-BZSNNMDCSA-N 0.000 description 1
- PYFNONMJYNJENN-AVGNSLFASA-N Lys-Lys-Gln Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N PYFNONMJYNJENN-AVGNSLFASA-N 0.000 description 1
- YUAXTFMFMOIMAM-QWRGUYRKSA-N Lys-Lys-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O YUAXTFMFMOIMAM-QWRGUYRKSA-N 0.000 description 1
- BXPHMHQHYHILBB-BZSNNMDCSA-N Lys-Lys-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BXPHMHQHYHILBB-BZSNNMDCSA-N 0.000 description 1
- AZOFEHCPMBRNFD-BZSNNMDCSA-N Lys-Phe-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCCN)C(O)=O)CC1=CC=CC=C1 AZOFEHCPMBRNFD-BZSNNMDCSA-N 0.000 description 1
- LUAJJLPHUXPQLH-KKUMJFAQSA-N Lys-Phe-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCCCN)N LUAJJLPHUXPQLH-KKUMJFAQSA-N 0.000 description 1
- SQXZLVXQXWILKW-KKUMJFAQSA-N Lys-Ser-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SQXZLVXQXWILKW-KKUMJFAQSA-N 0.000 description 1
- DIBZLYZXTSVGLN-CIUDSAMLSA-N Lys-Ser-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O DIBZLYZXTSVGLN-CIUDSAMLSA-N 0.000 description 1
- MIFFFXHMAHFACR-KATARQTJSA-N Lys-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CCCCN MIFFFXHMAHFACR-KATARQTJSA-N 0.000 description 1
- SQRLLZAQNOQCEG-KKUMJFAQSA-N Lys-Tyr-Ser Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CO)C(O)=O)CC1=CC=C(O)C=C1 SQRLLZAQNOQCEG-KKUMJFAQSA-N 0.000 description 1
- VWPJQIHBBOJWDN-DCAQKATOSA-N Lys-Val-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O VWPJQIHBBOJWDN-DCAQKATOSA-N 0.000 description 1
- QLFAPXUXEBAWEK-NHCYSSNCSA-N Lys-Val-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O QLFAPXUXEBAWEK-NHCYSSNCSA-N 0.000 description 1
- GILLQRYAWOMHED-DCAQKATOSA-N Lys-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN GILLQRYAWOMHED-DCAQKATOSA-N 0.000 description 1
- HMZPYMSEAALNAE-ULQDDVLXSA-N Lys-Val-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O HMZPYMSEAALNAE-ULQDDVLXSA-N 0.000 description 1
- BLIPQDLSCFGUFA-GUBZILKMSA-N Met-Arg-Asn Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O BLIPQDLSCFGUFA-GUBZILKMSA-N 0.000 description 1
- BMHIFARYXOJDLD-WPRPVWTQSA-N Met-Gly-Val Chemical compound [H]N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O BMHIFARYXOJDLD-WPRPVWTQSA-N 0.000 description 1
- WPTHAGXMYDRPFD-SRVKXCTJSA-N Met-Lys-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O WPTHAGXMYDRPFD-SRVKXCTJSA-N 0.000 description 1
- YLDSJJOGQNEQJK-AVGNSLFASA-N Met-Pro-Leu Chemical compound CSCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O YLDSJJOGQNEQJK-AVGNSLFASA-N 0.000 description 1
- RMLLCGYYVZKKRT-CIUDSAMLSA-N Met-Ser-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O RMLLCGYYVZKKRT-CIUDSAMLSA-N 0.000 description 1
- PHURAEXVWLDIGT-LPEHRKFASA-N Met-Ser-Pro Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N PHURAEXVWLDIGT-LPEHRKFASA-N 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 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
- HCTXJGRYAACKOB-SRVKXCTJSA-N Phe-Asn-Asp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N HCTXJGRYAACKOB-SRVKXCTJSA-N 0.000 description 1
- LDSOBEJVGGVWGD-DLOVCJGASA-N Phe-Asp-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 LDSOBEJVGGVWGD-DLOVCJGASA-N 0.000 description 1
- WMGVYPPIMZPWPN-SRVKXCTJSA-N Phe-Asp-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N WMGVYPPIMZPWPN-SRVKXCTJSA-N 0.000 description 1
- ZLGQEBCCANLYRA-RYUDHWBXSA-N Phe-Gly-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O ZLGQEBCCANLYRA-RYUDHWBXSA-N 0.000 description 1
- GXDPQJUBLBZKDY-IAVJCBSLSA-N Phe-Ile-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GXDPQJUBLBZKDY-IAVJCBSLSA-N 0.000 description 1
- LRBSWBVUCLLRLU-BZSNNMDCSA-N Phe-Leu-Lys Chemical compound CC(C)C[C@H](NC(=O)[C@@H](N)Cc1ccccc1)C(=O)N[C@@H](CCCCN)C(O)=O LRBSWBVUCLLRLU-BZSNNMDCSA-N 0.000 description 1
- ILGCZYGFYQLSDZ-KKUMJFAQSA-N Phe-Ser-His Chemical compound N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O ILGCZYGFYQLSDZ-KKUMJFAQSA-N 0.000 description 1
- JXQVYPWVGUOIDV-MXAVVETBSA-N Phe-Ser-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JXQVYPWVGUOIDV-MXAVVETBSA-N 0.000 description 1
- UNBFGVQVQGXXCK-KKUMJFAQSA-N Phe-Ser-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O UNBFGVQVQGXXCK-KKUMJFAQSA-N 0.000 description 1
- IPFXYNKCXYGSSV-KKUMJFAQSA-N Phe-Ser-Lys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N IPFXYNKCXYGSSV-KKUMJFAQSA-N 0.000 description 1
- YUPRIZTWANWWHK-DZKIICNBSA-N Phe-Val-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N YUPRIZTWANWWHK-DZKIICNBSA-N 0.000 description 1
- JTKGCYOOJLUETJ-ULQDDVLXSA-N Phe-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 JTKGCYOOJLUETJ-ULQDDVLXSA-N 0.000 description 1
- 108700001094 Plant Genes Proteins 0.000 description 1
- 241001533393 Potyviridae Species 0.000 description 1
- IWNOFCGBMSFTBC-CIUDSAMLSA-N Pro-Ala-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IWNOFCGBMSFTBC-CIUDSAMLSA-N 0.000 description 1
- VCYJKOLZYPYGJV-AVGNSLFASA-N Pro-Arg-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O VCYJKOLZYPYGJV-AVGNSLFASA-N 0.000 description 1
- KDIIENQUNVNWHR-JYJNAYRXSA-N Pro-Arg-Phe Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O KDIIENQUNVNWHR-JYJNAYRXSA-N 0.000 description 1
- CJZTUKSFZUSNCC-FXQIFTODSA-N Pro-Asp-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 CJZTUKSFZUSNCC-FXQIFTODSA-N 0.000 description 1
- VPFGPKIWSDVTOY-SRVKXCTJSA-N Pro-Glu-His Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O VPFGPKIWSDVTOY-SRVKXCTJSA-N 0.000 description 1
- CPRLKHJUFAXVTD-ULQDDVLXSA-N Pro-Leu-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CPRLKHJUFAXVTD-ULQDDVLXSA-N 0.000 description 1
- ABSSTGUCBCDKMU-UWVGGRQHSA-N Pro-Lys-Gly Chemical compound NCCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H]1CCCN1 ABSSTGUCBCDKMU-UWVGGRQHSA-N 0.000 description 1
- 108010003201 RGH 0205 Proteins 0.000 description 1
- 108020005091 Replication Origin Proteins 0.000 description 1
- 101710097247 Ribulose bisphosphate carboxylase large chain Proteins 0.000 description 1
- 101710104360 Ribulose bisphosphate carboxylase large chain, chromosomal Proteins 0.000 description 1
- 241000192026 Ruminococcus flavefaciens Species 0.000 description 1
- LVVBAKCGXXUHFO-ZLUOBGJFSA-N Ser-Ala-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(O)=O LVVBAKCGXXUHFO-ZLUOBGJFSA-N 0.000 description 1
- WTUJZHKANPDPIN-CIUDSAMLSA-N Ser-Ala-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N WTUJZHKANPDPIN-CIUDSAMLSA-N 0.000 description 1
- VGNYHOBZJKWRGI-CIUDSAMLSA-N Ser-Asn-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CO VGNYHOBZJKWRGI-CIUDSAMLSA-N 0.000 description 1
- QKQDTEYDEIJPNK-GUBZILKMSA-N Ser-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CO QKQDTEYDEIJPNK-GUBZILKMSA-N 0.000 description 1
- WBINSDOPZHQPPM-AVGNSLFASA-N Ser-Glu-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CO)N)O WBINSDOPZHQPPM-AVGNSLFASA-N 0.000 description 1
- KDGARKCAKHBEDB-NKWVEPMBSA-N Ser-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CO)N)C(=O)O KDGARKCAKHBEDB-NKWVEPMBSA-N 0.000 description 1
- YIUWWXVTYLANCJ-NAKRPEOUSA-N Ser-Ile-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O YIUWWXVTYLANCJ-NAKRPEOUSA-N 0.000 description 1
- DJACUBDEDBZKLQ-KBIXCLLPSA-N Ser-Ile-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O DJACUBDEDBZKLQ-KBIXCLLPSA-N 0.000 description 1
- JWOBLHJRDADHLN-KKUMJFAQSA-N Ser-Leu-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JWOBLHJRDADHLN-KKUMJFAQSA-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
- VGQVAVQWKJLIRM-FXQIFTODSA-N Ser-Ser-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O VGQVAVQWKJLIRM-FXQIFTODSA-N 0.000 description 1
- ZVBCMFDJIMUELU-BZSNNMDCSA-N Ser-Tyr-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CO)N ZVBCMFDJIMUELU-BZSNNMDCSA-N 0.000 description 1
- 101000910035 Streptococcus pyogenes serotype M1 CRISPR-associated endonuclease Cas9/Csn1 Proteins 0.000 description 1
- LVHHEVGYAZGXDE-KDXUFGMBSA-N Thr-Ala-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(=O)O)N)O LVHHEVGYAZGXDE-KDXUFGMBSA-N 0.000 description 1
- UKBSDLHIKIXJKH-HJGDQZAQSA-N Thr-Arg-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O UKBSDLHIKIXJKH-HJGDQZAQSA-N 0.000 description 1
- OJRNZRROAIAHDL-LKXGYXEUSA-N Thr-Asn-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O OJRNZRROAIAHDL-LKXGYXEUSA-N 0.000 description 1
- ZLNWJMRLHLGKFX-SVSWQMSJSA-N Thr-Cys-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O ZLNWJMRLHLGKFX-SVSWQMSJSA-N 0.000 description 1
- UBDDORVPVLEECX-FJXKBIBVSA-N Thr-Gly-Met Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O UBDDORVPVLEECX-FJXKBIBVSA-N 0.000 description 1
- MSIYNSBKKVMGFO-BHNWBGBOSA-N Thr-Gly-Pro Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N1CCC[C@@H]1C(=O)O)N)O MSIYNSBKKVMGFO-BHNWBGBOSA-N 0.000 description 1
- IMDMLDSVUSMAEJ-HJGDQZAQSA-N Thr-Leu-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IMDMLDSVUSMAEJ-HJGDQZAQSA-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
- SPVHQURZJCUDQC-VOAKCMCISA-N Thr-Lys-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O SPVHQURZJCUDQC-VOAKCMCISA-N 0.000 description 1
- UXUAZXWKIGPUCH-RCWTZXSCSA-N Thr-Met-Met Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCSC)C(O)=O UXUAZXWKIGPUCH-RCWTZXSCSA-N 0.000 description 1
- CGCMNOIQVAXYMA-UNQGMJICSA-N Thr-Met-Phe Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O CGCMNOIQVAXYMA-UNQGMJICSA-N 0.000 description 1
- WVVOFCVMHAXGLE-LFSVMHDDSA-N Thr-Phe-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(O)=O WVVOFCVMHAXGLE-LFSVMHDDSA-N 0.000 description 1
- BBPCSGKKPJUYRB-UVOCVTCTSA-N Thr-Thr-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O BBPCSGKKPJUYRB-UVOCVTCTSA-N 0.000 description 1
- NLWDSYKZUPRMBJ-IEGACIPQSA-N Thr-Trp-Leu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC(C)C)C(=O)O)N)O NLWDSYKZUPRMBJ-IEGACIPQSA-N 0.000 description 1
- ILUOMMDDGREELW-OSUNSFLBSA-N Thr-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)[C@@H](C)O ILUOMMDDGREELW-OSUNSFLBSA-N 0.000 description 1
- BPGDJSUFQKWUBK-KJEVXHAQSA-N Thr-Val-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 BPGDJSUFQKWUBK-KJEVXHAQSA-N 0.000 description 1
- XZSJDSBPEJBEFZ-QRTARXTBSA-N Trp-Asn-Val Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O XZSJDSBPEJBEFZ-QRTARXTBSA-N 0.000 description 1
- TVOGEPLDNYTAHD-CQDKDKBSSA-N Tyr-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 TVOGEPLDNYTAHD-CQDKDKBSSA-N 0.000 description 1
- DXYWRYQRKPIGGU-BPNCWPANSA-N Tyr-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 DXYWRYQRKPIGGU-BPNCWPANSA-N 0.000 description 1
- AYHSJESDFKREAR-KKUMJFAQSA-N Tyr-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AYHSJESDFKREAR-KKUMJFAQSA-N 0.000 description 1
- IXTQGBGHWQEEDE-AVGNSLFASA-N Tyr-Asp-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 IXTQGBGHWQEEDE-AVGNSLFASA-N 0.000 description 1
- NLMXVDDEQFKQQU-CFMVVWHZSA-N Tyr-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NLMXVDDEQFKQQU-CFMVVWHZSA-N 0.000 description 1
- WEFIPBYPXZYPHD-HJPIBITLSA-N Tyr-Cys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC1=CC=C(C=C1)O)N WEFIPBYPXZYPHD-HJPIBITLSA-N 0.000 description 1
- WPXKRJVHBXYLDT-JUKXBJQTSA-N Tyr-His-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CC2=CC=C(C=C2)O)N WPXKRJVHBXYLDT-JUKXBJQTSA-N 0.000 description 1
- USYGMBIIUDLYHJ-GVARAGBVSA-N Tyr-Ile-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 USYGMBIIUDLYHJ-GVARAGBVSA-N 0.000 description 1
- NXRGXTBPMOGFID-CFMVVWHZSA-N Tyr-Ile-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O NXRGXTBPMOGFID-CFMVVWHZSA-N 0.000 description 1
- HHFMNAVFGBYSAT-IGISWZIWSA-N Tyr-Ile-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N HHFMNAVFGBYSAT-IGISWZIWSA-N 0.000 description 1
- HFJJDMOFTCQGEI-STECZYCISA-N Tyr-Ile-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N HFJJDMOFTCQGEI-STECZYCISA-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
- QHLIUFUEUDFAOT-MGHWNKPDSA-N Tyr-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=CC=C(C=C1)O)N QHLIUFUEUDFAOT-MGHWNKPDSA-N 0.000 description 1
- OLYXUGBVBGSZDN-ACRUOGEOSA-N Tyr-Leu-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 OLYXUGBVBGSZDN-ACRUOGEOSA-N 0.000 description 1
- GYKDRHDMGQUZPU-MGHWNKPDSA-N Tyr-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC1=CC=C(C=C1)O)N GYKDRHDMGQUZPU-MGHWNKPDSA-N 0.000 description 1
- ZOBLBMGJKVJVEV-BZSNNMDCSA-N Tyr-Lys-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)O)N)O ZOBLBMGJKVJVEV-BZSNNMDCSA-N 0.000 description 1
- AVFGBGGRZOKSFS-KJEVXHAQSA-N Tyr-Met-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CC1=CC=C(C=C1)O)N)O AVFGBGGRZOKSFS-KJEVXHAQSA-N 0.000 description 1
- IGXLNVIYDYONFB-UFYCRDLUSA-N Tyr-Phe-Arg Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)C1=CC=C(O)C=C1 IGXLNVIYDYONFB-UFYCRDLUSA-N 0.000 description 1
- OKDNSNWJEXAMSU-IRXDYDNUSA-N Tyr-Phe-Gly Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)NCC(O)=O)C1=CC=C(O)C=C1 OKDNSNWJEXAMSU-IRXDYDNUSA-N 0.000 description 1
- JQOMHZMWQHXALX-FHWLQOOXSA-N Tyr-Tyr-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JQOMHZMWQHXALX-FHWLQOOXSA-N 0.000 description 1
- KRXFXDCNKLANCP-CXTHYWKRSA-N Tyr-Tyr-Ile Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 KRXFXDCNKLANCP-CXTHYWKRSA-N 0.000 description 1
- VKYDVKAKGDNZED-STECZYCISA-N Tyr-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC1=CC=C(C=C1)O)N VKYDVKAKGDNZED-STECZYCISA-N 0.000 description 1
- DDRBQONWVBDQOY-GUBZILKMSA-N Val-Ala-Arg Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O DDRBQONWVBDQOY-GUBZILKMSA-N 0.000 description 1
- UEOOXDLMQZBPFR-ZKWXMUAHSA-N Val-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N UEOOXDLMQZBPFR-ZKWXMUAHSA-N 0.000 description 1
- AUMNPAUHKUNHHN-BYULHYEWSA-N Val-Asn-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N AUMNPAUHKUNHHN-BYULHYEWSA-N 0.000 description 1
- HURRXSNHCCSJHA-AUTRQRHGSA-N Val-Gln-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N HURRXSNHCCSJHA-AUTRQRHGSA-N 0.000 description 1
- VVZDBPBZHLQPPB-XVKPBYJWSA-N Val-Glu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O VVZDBPBZHLQPPB-XVKPBYJWSA-N 0.000 description 1
- FEFZWCSXEMVSPO-LSJOCFKGSA-N Val-His-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](C)C(O)=O FEFZWCSXEMVSPO-LSJOCFKGSA-N 0.000 description 1
- BZMIYHIJVVJPCK-QSFUFRPTSA-N Val-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N BZMIYHIJVVJPCK-QSFUFRPTSA-N 0.000 description 1
- VXDSPJJQUQDCKH-UKJIMTQDSA-N Val-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N VXDSPJJQUQDCKH-UKJIMTQDSA-N 0.000 description 1
- RQOMPQGUGBILAG-AVGNSLFASA-N Val-Met-Leu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O RQOMPQGUGBILAG-AVGNSLFASA-N 0.000 description 1
- WMRWZYSRQUORHJ-YDHLFZDLSA-N Val-Phe-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)O)C(=O)O)N WMRWZYSRQUORHJ-YDHLFZDLSA-N 0.000 description 1
- QIVPZSWBBHRNBA-JYJNAYRXSA-N Val-Pro-Phe Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1ccccc1)C(O)=O QIVPZSWBBHRNBA-JYJNAYRXSA-N 0.000 description 1
- VIKZGAUAKQZDOF-NRPADANISA-N Val-Ser-Glu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O VIKZGAUAKQZDOF-NRPADANISA-N 0.000 description 1
- AOILQMZPNLUXCM-AVGNSLFASA-N Val-Val-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN AOILQMZPNLUXCM-AVGNSLFASA-N 0.000 description 1
- -1 ZFNs Proteins 0.000 description 1
- 108010017070 Zinc Finger Nucleases Proteins 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 108010045350 alanyl-tyrosyl-alanine Proteins 0.000 description 1
- 108010041407 alanylaspartic acid Proteins 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010069926 arginyl-glycyl-serine Proteins 0.000 description 1
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 108010010430 asparagine-proline-alanine Proteins 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 108010069495 cysteinyltyrosine Proteins 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 238000012214 genetic breeding Methods 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010080575 glutamyl-aspartyl-alanine Proteins 0.000 description 1
- 108010077435 glycyl-phenylalanyl-glycine Proteins 0.000 description 1
- 108010010147 glycylglutamine Proteins 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 108010028295 histidylhistidine Proteins 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 108010057952 lysyl-phenylalanyl-lysine Proteins 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 108091027963 non-coding RNA Proteins 0.000 description 1
- 102000042567 non-coding RNA Human genes 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 108010074082 phenylalanyl-alanyl-lysine Proteins 0.000 description 1
- 108010072637 phenylalanyl-arginyl-phenylalanine Proteins 0.000 description 1
- 108010084572 phenylalanyl-valine Proteins 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108010053725 prolylvaline Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 108010015840 seryl-prolyl-lysyl-lysine Proteins 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 108010001055 thymocartin Proteins 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 108010027345 wheylin-1 peptide Proteins 0.000 description 1
Images
Classifications
-
- 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
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
- C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
-
- 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
- C12N15/66—General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
-
- 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
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
- C12N15/8283—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for virus resistance
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/22—Ribonucleases RNAses, DNAses
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Virology (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用，包括pCambia1300‑RfxCas13d重组载体，构成如下：CaMV 35S启动子，潮霉素基因，CaMV poly(A)信号终止子，pVS1 RepA，pVS1复制起点，CaMV 35S启动子，RfxCas13d基因，NOS终止子；其中，RfxCas13d基因序列如SEQ ID No.1所示，NOS终止子序列如SEQ ID No.2所示。本发明利用带有GFP荧光标签的芜菁花叶病毒(TuMV‑GFP)侵染性克隆来指示RNA病毒自然侵害植物，利用RfxCas13d蛋白和sgRNA结合可以定向特定位置切割RNA。本发明构建的载体能够有效抑制TuMV的侵染，为植物病毒病的防控提供新的策略，具有重要的生产意义。
Description
技术领域
本发明涉及生物技术领域，特别是涉及一种高效定点编辑的CRISPR/RfxCas13d抗植物病毒载体及其构建方法和应用。
背景技术
基因组定点编辑技术是一种有效高质技术手段，可以用于植物功能基因组研究和作物分子遗传育种。基因组编辑技术主要通过以下三种人工内切核酸酶分别为：锌指核酸酶ZFNs、类转录激活因子效应物核酸酶TALENs和基于CRISPR/Cas***的由RNA引导的内切酶。针对DNA的基因编辑过程是在基因组靶位点处产生双链DNA断裂(DSBs)，DSBs可以通过非同源末端连接(NHEJ)和同源重组(HDR)进行修复。在CRISPR-Cas***的RNA引导内切核酸酶***中的内切核酸酶，如SpCas9和LbCpf1，已被证明是用于植物基因编辑和调控的多功能工具酶。RNA基因编辑是近些年在发现新型Cas蛋白(主要是Cas13蛋白)发展起来的。RNA一旦被切割之后，直接被降解。
在自然界中，植物病毒侵染植物的现象相当普遍。植物病毒可以感染许多重要的农作物，是影响农业生产的重要因素之一。植物病毒在世界各地广泛分布，除一小部分是DNA病毒外，大部分的植物病毒基因组是单链RNA(ssRNA)病毒。其中，芜菁花叶病毒(Turnipmosaic virus,TuMV)是单链、正义的植物RNA病毒，属于马铃薯Y病毒科病毒。TuMV能够侵染十子花科多种蔬菜作物与油料作物，是我国及世界农业生产上重要的病原物。目前，无有效的手段防治TuMV的侵染。
发明内容
本发明的目的是提供一种CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用。
本发明利用基因编辑技术直接靶向TuMV的RNA序列，导致目标RNA的切割和降解，实现抗病毒应用。本发明使用了来自Ruminococcus flavefaciens菌株strain XPD3002的细菌蛋白RfxCas13d蛋白，开发了基于RfxCas13d***的CRISPR植物基因组编辑工具。这种工具可以更高效切割RNA、更广泛地识别特异基因组序列，是一种高效地抗植物RNA病毒的工具。
一种CRISPR/RfxCas13d抗植物RNA病毒载体，包括pCambia1300-RfxCas13d重组载体，构成如下：CaMV 35S启动子，潮霉素基因，CaMV poly(A)信号终止子，pVS1 RepA，pVS1复制起点，CaMV 35S启动子，RfxCas13d基因，NOS终止子；
其中，RfxCas13d基因序列如SEQ ID No.1所示，NOS终止子序列如SEQ ID No.2所示；RfxCas13d编码的氨基酸序列如SEQ ID No.3所示。
pCambia1300-RfxCas13d重组载体的构建方法具体为，人工合成3003bp的RfxCas13d基因片段，并将其克隆至pUC57上，命名为pUC57：RfxCas13d；利用BamHI和NcoI对质粒pUC57：RfxCas13d和pCambia1300-spCas9进行双酶切，分别回收3003bp RfxCas13d基因片段和约10kb的pCambia1300载体骨架，利用T4 DNA连接酶将3003bp的RfxCas13d片段连接至pCambia1300载体骨架，质粒命名为pCambia-RfxCas13d；质粒测序后保存，验证成功后转农杆菌保存。
质粒pCambia1300-RfxCas13d-crRNA的构成如下：CaMV 35S启动子，潮霉素基因，CaMV poly(A)信号终止子，pVS1 RepA，pVS1复制起点，CaMV 35S启动子，RfxCas13d基因，NOS终止子，AtU6启动子，crRNA重复前导序列，含有两个BsaI酶切位点的spacer核苷酸序列，(T)8终止序列。
pCambia1300-RfxCas13d-crRNA重组载体的构建方法具体为，人工合成377bp的AtU6-crRNA基因片段，如SEQ ID No.4所示，并将其克隆至pUC57上，命名为pUC57：AtU6-crRNA；首先用HindIII和SbfI对pUC57：AtU6-crRNA和pCambia1300-RfxCas13d进行双酶切，分别回收377bp的片段和12.5kb的线性化载体骨架，利用T4 DNA连接酶将3003bp的crRNA片段连接至pCambia1300-RfxCas13d，命名为pCambia1300-RfxCas13d-crRNA。
CRISPR/RfxCas13d抗植物RNA病毒载体的应用，pCambia1300-RfxCas13d-crRNA根据目标进行设计靶标。以crGFP为例，引物crGFP-F1(如SEQ ID No.5所示)与crGFP-R1(如SEQ ID No.6所示)磷酸化后退火连接到pCambia1300-RfxCas13d-crRNA，命名为pCambia1300-RfxCas13d-crGFP。
CRISPR/RfxCas13d抗植物RNA病毒载体在用于将本氏烟基因组，和/或植物RNA病毒基因组中的内源基因沉默，RNA病毒外源RNA沉默中的至少一种的应用。
进一步的，CRISPR/RfxCas13d抗植物RNA病毒载体在抵抗芜菁花叶病毒中的应用。
同现有技术相比，本发明的突出效果在于：
TuMV是严重危害农业生产的病原物，目前无有效的防治手段。本发明利用侵染性克隆TuMV-GFP来指示RNA病毒自然侵害植物，利用RfxCas13d蛋白和sgRNA表达载体能够结合并切割特定位置的病毒RNA。本发明所构建的载体能够有效抑制TuMV的侵染，为植物病毒病的防控提供新的策略，具有重要的生产意义。
本发明构建出的重组载体pCambia1300-RfxCas13d具有以下特征和优势：
(1)细菌蛋白RfxCas13d不会影响植物正常生理活动；
(2)能够表达出特异的Cas13d蛋白与sgRNA结合，可用于切割某些特定的RNA上的序列，实现精准打靶；
(3)Cas13d可以切割RNA，因而不仅仅是植物内源产生的mRNA还是病毒基因组RNA都能实现定点切割。
下面结合附图说明和具体实施例对本发明所述的CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用作进一步说明。
附图说明
图1为pCambia1300-RfxCas13d(Cas13d)的载体示意图。
图2为pCambia1300-RfxCas13d-crRNA(Cas13d-crRNA)的载体示意图。
图3为TuMV-GFP侵染性克隆的病毒基因组，其中深灰色框为绿色荧光蛋白GFP。
图4为利用pCambia1300-RfxCas13d-crGFP进行抑制TuMV-GFP的侵染与TuMV衣壳蛋白(CP)蛋白积累量的分析。
(A)共接种TuMV-GFP侵染性克隆与pCambia1300-RfxCas13d-crGFP(简称Cas13d-crGFP，下同)或与Cas13d到本氏烟植物上，使用手提式紫外灯拍摄接种本氏烟植物10天后的照片。
(B)利用Western blot分析Cas13d-crGFP相比Cas13d对TuMV中CP蛋白积累量的分析情况，丽春红染色的Rubisco大亚基作为上样对照，表明Cas13d-crGFP能够有效降低TuMV中CP的积累量。
具体实施方式
实施例1CRISPR/RfxCas13d抗植物RNA病毒载体的构建
构建载体的技术路线如下：
1、pCambia1300-RfxCas13d重组质粒构建
人工合成3003bp的RfxCas13d基因片段，并将其克隆至pUC57上，命名为pUC57：RfxCas13d(生工生物工程(上海)股份有限公司)。利用BamHI和NcoI对质粒pUC57：RfxCas13d和pCambia1300-spCas9(购自杭州百格生物技术有限公司)进行双酶切，分别回收3003bp RfxCas13d基因片段和约10kb的pCambia1300载体骨架，利用T4 DNA连接酶(Thermo Cat#EL0011)将3003bp的RfxCas13d片段连接至pCambia1300载体骨架，质粒命名为pCambia-RfxCas13d。质粒测序后保存，验证成功以后转农杆菌保存。
如图1所示，质粒pCambia1300-RfxCas13d的构成如下：CaMV 35S启动子，潮霉素基因，CaMV poly(A)信号终止子，pVS1 RepA，pVS1复制起点，CaMV 35S启动子，RfxCas13d基因(如SEQ ID No.1所示)，NOS终止子(如SEQ ID No.2所示)。其中，RfxCas13d编码的氨基酸序列如SEQ ID No.3所示。
2、pCambia1300-RfxCas13d-crRNA重组质粒构建
人工合成377bp的AtU6-crRNA(如SEQ ID No.4所示)基因片段，并将其克隆至pUC57上，命名为pUC57：AtU6-crRNA(生工生物工程(上海)股份有限公司)。首先用HindIII和SbfI对pUC57：AtU6-crRNA和pCambia1300-RfxCas13d进行双酶切，分别回收377bp的片段和12.5kb的线性化载体骨架，利用T4 DNA连接酶(Thermo Cat#EL0011)将3003bp的crRNA片段连接至pCambia1300-RfxCas13d，命名为pCambia1300-RfxCas13d-crRNA。
如图2所示，质粒pCambia1300-RfxCas13d-crRNA的构成如下：CaMV 35S启动子，潮霉素基因，CaMV poly(A)信号终止子，pVS1 RepA，pVS1复制起点，CaMV 35S启动子，RfxCas13d基因，NOS终止子，AtU6启动子，crRNA重复前导序列，含有两个BsaI酶切位点的spacer核苷酸序列，(T)8终止序列。
以crGFP为例，引物crGFP-F1(如SEQ ID No.5所示)与crGFP-R1(如SEQ ID No.6所示)磷酸化后退火连接到pCambia1300-RfxCas13d-crRNA，命名为pCambia1300-RfxCas13d-crGFP。质粒测序后保存，验证成功以后转农杆菌保存。
其中，实施例1中的载体2是在载体1CRISPR/RfxCas13d的***中加入sgRNA，即在pCambia1300-RfxCas13d载体骨架中加入sgRNA表达框，只有有了sgRNA的指导作用，Cas13d蛋白才能正确识别指定位点RNA发挥功能。
实施例2：利用pCambia1300-RfxCas13d-crGFP能抑制TuMV-GFP的侵染
1、农杆菌植物侵染及接种
(1)活化农杆菌，接种含有pCambia1300-RfxCas13d(简称Cas13d)，pCambia1300-RfxCas13d-crGFP(简称Cas13d-crGFP)，TuMV-GFP质粒的农杆菌菌液培养至OD600约为1.5时，4,000rpm离心12min收集菌体，用含终浓度为10mM氯化镁，10mM MES(pH5.6)和100mM乙酰丁香酮的双蒸水溶液(浸润缓冲液)重悬菌体，调整菌液浓度使其OD600约为1.0左右，室温下放置3小时。同时注射Cas13d-crGFP与TuMV-GFP农杆菌的侵染性克隆，或Cas13d-crGFP与TuMV-GFP农杆菌的侵染性克隆(图3为TuMV-GFP的载体示意图)，菌液浓度不能过高，OD600在0.4至0.6为宜；
(2)注射接种，用1ml的注射器刺到本氏烟叶片背面2-4个小孔，一手按住叶面小孔处，一手用无针头的针管将菌液沿小孔慢慢注入叶片组织空隙处，至整个叶片全部被浸满；
(3)浸润后的植物于25℃温室中培养，光照周期为16/8小时(光照/黑暗)。观察植物症状，确定TuMV-GFP发病情况，手持UV灯下观察，接种对照组Cas13d(非靶标载体对照)与TuMV-GFP的绿色荧光蛋白表达情况。结果如图3所示。
2、紫外灯拍照观察，结果如图4所示。
以上结果说明Cas13d-crGFP能够对TuMV-GFP有效干扰，阻遏RNA病毒的扩增，抑制病毒的侵染和病毒蛋白的积累。
以上所述的实施例仅仅是对本发明的优选实施方式进行描述，并非对本发明的范围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发明的技术方案作出的各种变形和改进，均应落入本发明权利要求书确定的保护范围内。
序列表
<110> 中国农业科学院植物保护研究所
<120> CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 3003
<212> DNA
<213> NLS-RfxCas13d-NLS(Artificial Sequence)
<400> 1
atgtctccga aaaaaaaaag gaaggtcgag gcgagcattg agaaaaagaa gagctttgct 60
aaaggtatgg gggttaagag tacattggtg tccggttcca aagtgtatat gactacattt 120
gcggaaggca gcgacgcaag gctggagaaa atagttgaag gcgattcaat aagatcggtg 180
aatgaaggag aagccttctc cgctgaaatg gccgacaaaa atgccggtta taagattggt 240
aacgctaagt tctcgcaccc aaagggttat gcagtcgtgg cgaacaatcc actgtacaca 300
ggtcctgtgc agcaggatat gcttggtttg aaggaaacgt tggaaaagcg gtacttcggt 360
gagtccgctg acggcaatga taatatatgt attcaggtga tacacaacat cctcgatata 420
gaaaaaatcc tcgctgagta cattacaaat gccgcctacg cggtcaataa catatccggt 480
ttggacaaag acatcatagg atttggaaaa ttctctacag tgtacacgta cgatgaattt 540
aaagacccgg agcatcacag agcagctttc aacaacaatg ataagctcat aaatgcaata 600
aaagcgcaat atgatgaatt tgacaacttt cttgataacc cgaggctggg ctattttggt 660
caggctttct tcagcaaaga agggcgcaac tatattataa attacggcaa tgagtgctat 720
gatattttgg cattgttgtc cggactgcgg cattgggttg ttcataataa cgaggaagag 780
agtaggatca gtcgcacatg gctgtataac cttgataaga atctcgataa tgagtatatt 840
tcgactttga actacttgta tgatcgcatt actaacgaac tcaccaacag cttttcgaag 900
aactcggcag ccaacgtgaa ttatatagcg gaaaccttgg ggattaaccc agcagaattc 960
gcagaacagt actttcgctt cagcataatg aaggaacaaa agaacctcgg ttttaacatc 1020
acaaaactca gagaggttat gctggataga aaagacatgt cagaaattcg gaaaaatcat 1080
aaagtgttcg attccatcag gacgaaggtc tacacaatga tggatttcgt catctacaga 1140
tattatattg aggaggatgc aaaagtcgca gcggccaaca aaagcctccc tgacaatgaa 1200
aagtcgctct ctgaaaagga catctttgtc ataaaccttc ggggcagttt taacgatgac 1260
caaaaggacg ccttgtacta cgatgaagca aatcgcatct ggaggaaact tgagaacata 1320
atgcataaca taaaggagtt tcgggggaac aagacgagag aatacaaaaa gaaggacgcg 1380
ccaagacttc ctagaattct cccagcgggg cgcgacgtct cagcgttctc caagctcatg 1440
tacgcgctta ccatgttcct cgatggaaaa gagataaatg atcttttgac tacgctcatt 1500
aacaagttcg acaacattca atctttcctg aaagtgatgc ctctcatagg ggtcaacgca 1560
aagttcgttg aagaatacgc ctttttcaag gactctgcga agatagccga tgaactccgc 1620
ctcataaaga gctttgcgcg gatgggtgaa cctattgctg acgcccggag ggcaatgtat 1680
attgacgcga tcaggattct tggaactaat ctctcctacg acgaacttaa ggctcttgct 1740
gatacctttt ctcttgatga aaacgggaac aaactcaaga agggaaaaca cggtatgcgg 1800
aatttcatca taaataacgt tatttcaaac aagagattcc attacctgat aagatacgga 1860
gatccagccc atctgcacga aatcgcgaaa aacgaggctg ttgttaaatt cgttttgggg 1920
agaatcgctg acatacaaaa aaagcagggg caaaacggga agaaccagat cgaccggtac 1980
tacgaaacct gtatcggtaa ggacaaaggg aagagtgtgt ccgaaaaggt tgacgccctt 2040
acaaaaatca tcaccggtat gaactatgac caattcgaca agaaaagaag tgttatagaa 2100
gatacgggaa gagagaatgc ggagcgggaa aaatttaaaa agatcatatc gctctatctg 2160
accgttatct atcatatcct taaaaacata gtcaacatca acgcacggta cgtgataggc 2220
ttccattgtg tggaacggga cgcccagttg tacaaagaga aaggatacga cataaacctc 2280
aagaagctcg aagagaaggg ttttagctct gttacgaaac tttgtgcggg tatcgatgaa 2340
accgcgcctg acaaacggaa agacgttgaa aaggagatgg cagaacgcgc taaagagtct 2400
atagacagtc ttgagtcagc aaatcccaag ctctacgcga actacataaa atattctgac 2460
gaaaaaaaag ctgaagaatt taccagacaa ataaacagag agaaggctaa gactgcgttg 2520
aatgcctatc tgcggaacac taaatggaat gtcataattc gggaagacct tctgcggatc 2580
gacaataaaa cctgcaccct ctttagaaat aaggctgtcc acctggaagt tgctcgctat 2640
gtgcatgcgt atattaacga cattgctgag gttaacagct actttcagct gtatcattac 2700
atcatgcaga ggattattat gaacgagcgc tacgagaagt cctccgggaa ggtttcagag 2760
tattttgacg cagtcaacga tgagaaaaag tacaacgatc ggctgctgaa actcctgtgt 2820
gtcccattcg ggtattgtat accgcgcttc aagaacctct caatagaggc gctctttgac 2880
cgcaacgagg ccgcaaagtt tgataaagaa aaaaagaaag ttagtgggaa tagtggctca 2940
ggaccaaaga agaaaagaaa ggttgcagcg gcatatccgt acgatgtgcc ggattatgcg 3000
tga 3003
<210> 2
<211> 225
<212> DNA
<213> NOS terminator(Artificial Sequence)
<400> 2
gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg 60
atgattatca tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc 120
atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac 180
gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcg 225
<210> 3
<211> 1000
<212> PRT
<213> NLS-RfxCas13d-NLS(Artificial Sequence)
<400> 3
Met Ser Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Ile Glu Lys Lys
1 5 10 15
Lys Ser Phe Ala Lys Gly Met Gly Val Lys Ser Thr Leu Val Ser Gly
20 25 30
Ser Lys Val Tyr Met Thr Thr Phe Ala Glu Gly Ser Asp Ala Arg Leu
35 40 45
Glu Lys Ile Val Glu Gly Asp Ser Ile Arg Ser Val Asn Glu Gly Glu
50 55 60
Ala Phe Ser Ala Glu Met Ala Asp Lys Asn Ala Gly Tyr Lys Ile Gly
65 70 75 80
Asn Ala Lys Phe Ser His Pro Lys Gly Tyr Ala Val Val Ala Asn Asn
85 90 95
Pro Leu Tyr Thr Gly Pro Val Gln Gln Asp Met Leu Gly Leu Lys Glu
100 105 110
Thr Leu Glu Lys Arg Tyr Phe Gly Glu Ser Ala Asp Gly Asn Asp Asn
115 120 125
Ile Cys Ile Gln Val Ile His Asn Ile Leu Asp Ile Glu Lys Ile Leu
130 135 140
Ala Glu Tyr Ile Thr Asn Ala Ala Tyr Ala Val Asn Asn Ile Ser Gly
145 150 155 160
Leu Asp Lys Asp Ile Ile Gly Phe Gly Lys Phe Ser Thr Val Tyr Thr
165 170 175
Tyr Asp Glu Phe Lys Asp Pro Glu His His Arg Ala Ala Phe Asn Asn
180 185 190
Asn Asp Lys Leu Ile Asn Ala Ile Lys Ala Gln Tyr Asp Glu Phe Asp
195 200 205
Asn Phe Leu Asp Asn Pro Arg Leu Gly Tyr Phe Gly Gln Ala Phe Phe
210 215 220
Ser Lys Glu Gly Arg Asn Tyr Ile Ile Asn Tyr Gly Asn Glu Cys Tyr
225 230 235 240
Asp Ile Leu Ala Leu Leu Ser Gly Leu Arg His Trp Val Val His Asn
245 250 255
Asn Glu Glu Glu Ser Arg Ile Ser Arg Thr Trp Leu Tyr Asn Leu Asp
260 265 270
Lys Asn Leu Asp Asn Glu Tyr Ile Ser Thr Leu Asn Tyr Leu Tyr Asp
275 280 285
Arg Ile Thr Asn Glu Leu Thr Asn Ser Phe Ser Lys Asn Ser Ala Ala
290 295 300
Asn Val Asn Tyr Ile Ala Glu Thr Leu Gly Ile Asn Pro Ala Glu Phe
305 310 315 320
Ala Glu Gln Tyr Phe Arg Phe Ser Ile Met Lys Glu Gln Lys Asn Leu
325 330 335
Gly Phe Asn Ile Thr Lys Leu Arg Glu Val Met Leu Asp Arg Lys Asp
340 345 350
Met Ser Glu Ile Arg Lys Asn His Lys Val Phe Asp Ser Ile Arg Thr
355 360 365
Lys Val Tyr Thr Met Met Asp Phe Val Ile Tyr Arg Tyr Tyr Ile Glu
370 375 380
Glu Asp Ala Lys Val Ala Ala Ala Asn Lys Ser Leu Pro Asp Asn Glu
385 390 395 400
Lys Ser Leu Ser Glu Lys Asp Ile Phe Val Ile Asn Leu Arg Gly Ser
405 410 415
Phe Asn Asp Asp Gln Lys Asp Ala Leu Tyr Tyr Asp Glu Ala Asn Arg
420 425 430
Ile Trp Arg Lys Leu Glu Asn Ile Met His Asn Ile Lys Glu Phe Arg
435 440 445
Gly Asn Lys Thr Arg Glu Tyr Lys Lys Lys Asp Ala Pro Arg Leu Pro
450 455 460
Arg Ile Leu Pro Ala Gly Arg Asp Val Ser Ala Phe Ser Lys Leu Met
465 470 475 480
Tyr Ala Leu Thr Met Phe Leu Asp Gly Lys Glu Ile Asn Asp Leu Leu
485 490 495
Thr Thr Leu Ile Asn Lys Phe Asp Asn Ile Gln Ser Phe Leu Lys Val
500 505 510
Met Pro Leu Ile Gly Val Asn Ala Lys Phe Val Glu Glu Tyr Ala Phe
515 520 525
Phe Lys Asp Ser Ala Lys Ile Ala Asp Glu Leu Arg Leu Ile Lys Ser
530 535 540
Phe Ala Arg Met Gly Glu Pro Ile Ala Asp Ala Arg Arg Ala Met Tyr
545 550 555 560
Ile Asp Ala Ile Arg Ile Leu Gly Thr Asn Leu Ser Tyr Asp Glu Leu
565 570 575
Lys Ala Leu Ala Asp Thr Phe Ser Leu Asp Glu Asn Gly Asn Lys Leu
580 585 590
Lys Lys Gly Lys His Gly Met Arg Asn Phe Ile Ile Asn Asn Val Ile
595 600 605
Ser Asn Lys Arg Phe His Tyr Leu Ile Arg Tyr Gly Asp Pro Ala His
610 615 620
Leu His Glu Ile Ala Lys Asn Glu Ala Val Val Lys Phe Val Leu Gly
625 630 635 640
Arg Ile Ala Asp Ile Gln Lys Lys Gln Gly Gln Asn Gly Lys Asn Gln
645 650 655
Ile Asp Arg Tyr Tyr Glu Thr Cys Ile Gly Lys Asp Lys Gly Lys Ser
660 665 670
Val Ser Glu Lys Val Asp Ala Leu Thr Lys Ile Ile Thr Gly Met Asn
675 680 685
Tyr Asp Gln Phe Asp Lys Lys Arg Ser Val Ile Glu Asp Thr Gly Arg
690 695 700
Glu Asn Ala Glu Arg Glu Lys Phe Lys Lys Ile Ile Ser Leu Tyr Leu
705 710 715 720
Thr Val Ile Tyr His Ile Leu Lys Asn Ile Val Asn Ile Asn Ala Arg
725 730 735
Tyr Val Ile Gly Phe His Cys Val Glu Arg Asp Ala Gln Leu Tyr Lys
740 745 750
Glu Lys Gly Tyr Asp Ile Asn Leu Lys Lys Leu Glu Glu Lys Gly Phe
755 760 765
Ser Ser Val Thr Lys Leu Cys Ala Gly Ile Asp Glu Thr Ala Pro Asp
770 775 780
Lys Arg Lys Asp Val Glu Lys Glu Met Ala Glu Arg Ala Lys Glu Ser
785 790 795 800
Ile Asp Ser Leu Glu Ser Ala Asn Pro Lys Leu Tyr Ala Asn Tyr Ile
805 810 815
Lys Tyr Ser Asp Glu Lys Lys Ala Glu Glu Phe Thr Arg Gln Ile Asn
820 825 830
Arg Glu Lys Ala Lys Thr Ala Leu Asn Ala Tyr Leu Arg Asn Thr Lys
835 840 845
Trp Asn Val Ile Ile Arg Glu Asp Leu Leu Arg Ile Asp Asn Lys Thr
850 855 860
Cys Thr Leu Phe Arg Asn Lys Ala Val His Leu Glu Val Ala Arg Tyr
865 870 875 880
Val His Ala Tyr Ile Asn Asp Ile Ala Glu Val Asn Ser Tyr Phe Gln
885 890 895
Leu Tyr His Tyr Ile Met Gln Arg Ile Ile Met Asn Glu Arg Tyr Glu
900 905 910
Lys Ser Ser Gly Lys Val Ser Glu Tyr Phe Asp Ala Val Asn Asp Glu
915 920 925
Lys Lys Tyr Asn Asp Arg Leu Leu Lys Leu Leu Cys Val Pro Phe Gly
930 935 940
Tyr Cys Ile Pro Arg Phe Lys Asn Leu Ser Ile Glu Ala Leu Phe Asp
945 950 955 960
Arg Asn Glu Ala Ala Lys Phe Asp Lys Glu Lys Lys Lys Val Ser Gly
965 970 975
Asn Ser Gly Ser Gly Pro Lys Lys Lys Arg Lys Val Ala Ala Ala Tyr
980 985 990
Pro Tyr Asp Val Pro Asp Tyr Ala
995 1000
<210> 4
<211> 380
<212> DNA
<213> AtU6-crRNA(Artificial Sequence)
<400> 4
aagcttcatt cggagttttt gtatcttgtt tcatagtttg tcccaggatt agaatgatta 60
ggcatcgaac cttcaagaat ttgattgaat aaaacatctt cattcttaag atatgaagat 120
aatcttcaaa aggcccctgg gaatctgaaa gaagagaagc aggcccattt atatgggaaa 180
gaacaatagt atttcttata taggcccatt taagttgaaa acaatcttca aaagtcccac 240
atcgcttaga taagaaaacg aagctgagtt tatatacagc tagagtcgaa gtagtgattg 300
aacccctacc aactggtcgg ggtttgaaac agagaccaga attcggtctc attttttttt 360
ggatccacta gtcctgcagg 380
<210> 5
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
aaacaacagg tagttttcca gtagtg 26
<210> 6
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
aaaacactac tggaaaacta cctgtt 26
Claims (9)
1.一种CRISPR/RfxCas13d抗植物RNA病毒载体，其特征在于：包括pCambia1300-RfxCas13d重组载体，构成如下：CaMV 35S启动子，潮霉素基因，CaMV poly(A)信号终止子，pVS1 RepA，pVS1复制起点，CaMV 35S启动子，RfxCas13d基因，NOS终止子；
其中，RfxCas13d基因序列如SEQ ID No.1所示，NOS终止子序列如SEQ ID No.2所示。
2.根据权利要求1所述的CRISPR/RfxCas13d抗植物RNA病毒载体，其特征在于：
RfxCas13d编码的氨基酸序列如SEQ ID No.3所示。
3.根据权利要求1所述的CRISPR/RfxCas13d抗植物RNA病毒载体，其特征在于：还包括pCambia1300-RfxCas13d-crRNA重组载体，构成如下：CaMV 35S启动子，潮霉素基因，CaMVpoly(A)信号终止子，pVS1 RepA，pVS1复制起点，CaMV 35S启动子，RfxCas13d基因，NOS终止子，AtU6启动子，crRNA重复前导序列，含有两个BsaI酶切位点的spacer核苷酸序列，(T)8终止序列。
4.权利要求1-3任一所述的CRISPR/RfxCas13d抗植物RNA病毒载体的构建方法，其特征在于：pCambia1300-RfxCas13d重组载体的构建方法具体为，人工合成3003bp的RfxCas13d基因片段，并将其克隆至pUC57上，命名为pUC57：RfxCas13d；利用BamHI和NcoI对质粒pUC57：RfxCas13d和pCambia1300-spCas9进行双酶切，分别回收3003bp RfxCas13d基因片段和约10kb的pCambia1300载体骨架，利用T4 DNA连接酶将3003bp的RfxCas13d片段连接至pCambia1300载体骨架，质粒命名为pCambia-RfxCas13d；质粒测序后保存，验证成功后转农杆菌保存。
5.权利要求3所述的CRISPR/RfxCas13d抗植物RNA病毒载体的构建方法，其特征在于：pCambia1300-RfxCas13d-crRNA重组载体的构建方法具体为，人工合成377bp的AtU6-crRNA基因片段，如SEQ ID No.4所示，并将其克隆至pUC57上，命名为pUC57：AtU6-crRNA；首先用HindIII和SbfI对pUC57：AtU6-crRNA和pCambia1300-RfxCas13d进行双酶切，分别回收377bp的片段和12.5kb的线性化载体骨架，利用T4 DNA连接酶将3003bp的crRNA片段连接至pCambia1300-RfxCas13d，命名为pCambia1300-RfxCas13d-crRNA。
6.权利要求1-3任一所述的CRISPR/RfxCas13d抗植物RNA病毒载体的应用，其特征在于：所述pCambia1300-RfxCas13d-crRNA根据目标进行设计靶标。
7.根据权利要求6所述的CRISPR/RfxCas13d抗植物RNA病毒载体的应用，其特征在于：CRISPR/RfxCas13d抗植物RNA病毒载体在用于将本氏烟基因组，和/或植物RNA病毒基因组中的内源基因沉默，RNA病毒外源RNA沉默中的至少一种的应用。
8.根据权利要求8所述的CRISPR/RfxCas13d抗植物RNA病毒载体的应用，其特征在于：在抵抗芜菁花叶病毒中的应用。
9.根据权利要求9所述的CRISPR/RfxCas13d抗植物RNA病毒载体的应用，其特征在于：
1)将构建好的载体转化农杆菌，农杆菌鉴定阳性后浸润本氏烟植物叶片，同时接种芜菁花叶病毒的侵染性克隆，7天后观察***叶片的表型、发病症状并取样；
2)取样后进行分子检测，蛋白水平上的检测试验采用Western blot的方法检测病毒积累量的变化。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010443155.8A CN111549054A (zh) | 2020-05-22 | 2020-05-22 | CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010443155.8A CN111549054A (zh) | 2020-05-22 | 2020-05-22 | CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111549054A true CN111549054A (zh) | 2020-08-18 |
Family
ID=72008442
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010443155.8A Pending CN111549054A (zh) | 2020-05-22 | 2020-05-22 | CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111549054A (zh) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113249403A (zh) * | 2021-03-31 | 2021-08-13 | 江苏师范大学 | 一种具有spvd抗性甘薯的育种方法 |
| CN114990093A (zh) * | 2022-06-24 | 2022-09-02 | 吉林大学 | 氨基酸序列小的蛋白序列mini rfx-cas13d |
| CN116083400A (zh) * | 2022-12-08 | 2023-05-09 | 广州瑞风生物科技有限公司 | Cas蛋白截短体、构建其的方法及其应用 |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5919705A (en) * | 1993-06-04 | 1999-07-06 | Novartis Finance Corporation | Plant virus DNA constructs and virus resistant plants comprising said constructs |
| CN108753815A (zh) * | 2018-06-19 | 2018-11-06 | 东北农业大学 | 一种大通量筛选植物病毒复制必需寄主基因的方法 |
| US20190002889A1 (en) * | 2017-06-30 | 2019-01-03 | Arbor Biotechnologies, Inc. | Novel crispr rna targeting enzymes and systems and uses thereof |
| CN110066824A (zh) * | 2018-01-24 | 2019-07-30 | 中国农业科学院植物保护研究所 | 一套用于水稻的碱基编辑人工*** |
| CN110257420A (zh) * | 2019-06-14 | 2019-09-20 | 中国科学院武汉植物园 | 基于CasRx的植物基因沉默载体及其构建方法和应用 |
| CN110656123A (zh) * | 2019-10-12 | 2020-01-07 | 中国农业大学 | 基于CRISPR-Cas13d***的sgRNA高效作用靶点的筛选方法及应用 |
| WO2020160150A1 (en) * | 2019-01-29 | 2020-08-06 | The Regents Of The University Of California | Rna-targeting cas enzymes |
-
2020
- 2020-05-22 CN CN202010443155.8A patent/CN111549054A/zh active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5919705A (en) * | 1993-06-04 | 1999-07-06 | Novartis Finance Corporation | Plant virus DNA constructs and virus resistant plants comprising said constructs |
| US20190002889A1 (en) * | 2017-06-30 | 2019-01-03 | Arbor Biotechnologies, Inc. | Novel crispr rna targeting enzymes and systems and uses thereof |
| CN110066824A (zh) * | 2018-01-24 | 2019-07-30 | 中国农业科学院植物保护研究所 | 一套用于水稻的碱基编辑人工*** |
| CN108753815A (zh) * | 2018-06-19 | 2018-11-06 | 东北农业大学 | 一种大通量筛选植物病毒复制必需寄主基因的方法 |
| WO2020160150A1 (en) * | 2019-01-29 | 2020-08-06 | The Regents Of The University Of California | Rna-targeting cas enzymes |
| CN110257420A (zh) * | 2019-06-14 | 2019-09-20 | 中国科学院武汉植物园 | 基于CasRx的植物基因沉默载体及其构建方法和应用 |
| CN110656123A (zh) * | 2019-10-12 | 2020-01-07 | 中国农业大学 | 基于CRISPR-Cas13d***的sgRNA高效作用靶点的筛选方法及应用 |
Non-Patent Citations (4)
| Title |
|---|
| AHMED MAHAS等: "CRISPR-Cas13d mediates robust RNA virus interference in plants", 《GENOME BIOLOGY》 * |
| 无: "Accession. MH142259.1, Cloning vector pM2Z-fun, complete sequence", 《GENEBANK》 * |
| 毕登峰 等: "CRISPR/Cas13d介导猪胎儿成纤维细胞SUV39H1/SUV39H2基因敲降", 《中国畜牧兽医》 * |
| 郭振飞: "《牧草生物技术》", 31 January 2011, 北京：中国农业大学出版社 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113249403A (zh) * | 2021-03-31 | 2021-08-13 | 江苏师范大学 | 一种具有spvd抗性甘薯的育种方法 |
| CN113249403B (zh) * | 2021-03-31 | 2023-08-04 | 江苏师范大学 | 一种具有spvd抗性甘薯的育种方法 |
| CN114990093A (zh) * | 2022-06-24 | 2022-09-02 | 吉林大学 | 氨基酸序列小的蛋白序列mini rfx-cas13d |
| CN114990093B (zh) * | 2022-06-24 | 2024-02-13 | 吉林大学 | 氨基酸序列小的蛋白序列mini rfx-cas13d |
| CN116083400A (zh) * | 2022-12-08 | 2023-05-09 | 广州瑞风生物科技有限公司 | Cas蛋白截短体、构建其的方法及其应用 |
| CN116083400B (zh) * | 2022-12-08 | 2023-12-12 | 广州瑞风生物科技有限公司 | Cas蛋白截短体、构建其的方法及其应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111549054A (zh) | CRISPR/RfxCas13d抗植物RNA病毒载体及其构建方法和应用 | |
| CN109705198B (zh) | OsCKX7蛋白质及其编码基因在调控植物纹枯病抗性中的应用 | |
| CN109627304B (zh) | 小麦抗条锈病相关蛋白TaNAC3及其编码基因与应用 | |
| CN110028566B (zh) | GhPRXR1蛋白及其编码基因在调控棉籽含油量中的应用 | |
| CN112575003B (zh) | 本氏烟hakai基因在调控植物抗病毒中的应用及转基因植物培育方法 | |
| CN113430212B (zh) | 苹果砧木抗盐胁迫相关基因MdLysMe3及其编码蛋白与应用 | |
| US20210163973A1 (en) | Plant constitutive expression promoter and applications thereof | |
| CN111574605B (zh) | 水稻基因OsLAT5在调节敌草快的吸收积累中的应用 | |
| CN114317460B (zh) | 瓜蒌斑驳花叶病毒及其侵染性克隆载体、构建方法和应用 | |
| IL271283B2 (en) | Methods and compositions for targeted genomic insertion | |
| CN110713994B (zh) | 一种植物耐逆性相关蛋白TaMAPK3及其编码基因和应用 | |
| CN113234729B (zh) | 可显著提高棉花黄萎病抗性的基因GauRev2及其应用 | |
| CN111205357B (zh) | 小麦抗条锈病相关蛋白TaWLT14.2及其编码基因与应用 | |
| CN111206040B (zh) | 水稻白叶枯病抗性相关基因OsDuf6及其应用 | |
| CN112646011A (zh) | 一种与植物抗逆性相关的蛋白PHD-Finger17及其编码基因与应用 | |
| CN104404043A (zh) | 疣粒野生稻抗白叶枯病相关基因Me094启动子 | |
| CN114686456B (zh) | 基于双分子脱氨酶互补的碱基编辑***及其应用 | |
| CN112813092B (zh) | GbBCCP5蛋白质及其编码基因在调控生物油脂含量中的应用 | |
| CN114875062A (zh) | 一种通过基因组编辑提高小麦赤霉病抗性的方法 | |
| CN113881700A (zh) | 拟南芥AtSR33基因在抑制芜菁花叶病毒侵染中的应用 | |
| CN108504661B (zh) | 二球悬铃木cpc家族基因在调控植物表皮毛中的应用 | |
| CN110272915B (zh) | 一种通过基因编辑技术培育抗白叶枯病水稻的方法 | |
| CN111440232A (zh) | 小麦抗条锈病相关蛋白TaNTL1及其编码基因与应用 | |
| CN111635909B (zh) | 一个木瓜类半胱氨酸蛋白酶基因TaRD21A及其表达载体和应用 | |
| CN115927394B (zh) | 玉米VPS23类似基因ZmVPS23L及其应用 |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200818 |