CN114853857B - 调控玉米花粉育性和花粉耐热性的方法 - Google Patents
调控玉米花粉育性和花粉耐热性的方法 Download PDFInfo
- Publication number
- CN114853857B CN114853857B CN202210180293.0A CN202210180293A CN114853857B CN 114853857 B CN114853857 B CN 114853857B CN 202210180293 A CN202210180293 A CN 202210180293A CN 114853857 B CN114853857 B CN 114853857B
- Authority
- CN
- China
- Prior art keywords
- leu
- ala
- glu
- arg
- val
- 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
- 240000008042 Zea mays Species 0.000 title claims abstract description 156
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 154
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 78
- 235000005822 corn Nutrition 0.000 title claims abstract description 78
- 230000035558 fertility Effects 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 32
- 230000001105 regulatory effect Effects 0.000 title abstract description 4
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 160
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 94
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 14
- 206010021929 Infertility male Diseases 0.000 claims abstract description 12
- 208000007466 Male Infertility Diseases 0.000 claims abstract description 12
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 76
- 235000009973 maize Nutrition 0.000 claims description 76
- 125000003729 nucleotide group Chemical group 0.000 claims description 35
- 239000002773 nucleotide Substances 0.000 claims description 33
- 150000007523 nucleic acids Chemical class 0.000 claims description 32
- 108020004414 DNA Proteins 0.000 claims description 30
- 102000039446 nucleic acids Human genes 0.000 claims description 29
- 108020004707 nucleic acids Proteins 0.000 claims description 29
- 230000014509 gene expression Effects 0.000 claims description 23
- 239000013598 vector Substances 0.000 claims description 22
- 230000009261 transgenic effect Effects 0.000 claims description 20
- 102000053602 DNA Human genes 0.000 claims description 17
- 244000005700 microbiome Species 0.000 claims description 13
- 230000000694 effects Effects 0.000 claims description 12
- 108091026890 Coding region Proteins 0.000 claims description 6
- 238000009395 breeding Methods 0.000 claims description 6
- 230000001488 breeding effect Effects 0.000 claims description 6
- 230000002401 inhibitory effect Effects 0.000 claims description 5
- 208000000509 infertility Diseases 0.000 claims description 4
- 230000036512 infertility Effects 0.000 claims description 4
- 208000021267 infertility disease Diseases 0.000 claims description 4
- 108020001507 fusion proteins Proteins 0.000 claims description 3
- 102000037865 fusion proteins Human genes 0.000 claims description 3
- 230000001737 promoting effect Effects 0.000 claims description 2
- 241000254173 Coleoptera Species 0.000 claims 1
- 238000002474 experimental method Methods 0.000 abstract description 8
- 235000018102 proteins Nutrition 0.000 description 81
- 241000196324 Embryophyta Species 0.000 description 22
- 235000013339 cereals Nutrition 0.000 description 19
- 239000013612 plasmid Substances 0.000 description 16
- 230000035772 mutation Effects 0.000 description 13
- 241000589158 Agrobacterium Species 0.000 description 12
- 230000008642 heat stress Effects 0.000 description 11
- 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 10
- 125000000539 amino acid group Chemical group 0.000 description 10
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 8
- SITWEMZOJNKJCH-UHFFFAOYSA-N L-alanine-L-arginine Natural products CC(N)C(=O)NC(C(O)=O)CCCNC(N)=N SITWEMZOJNKJCH-UHFFFAOYSA-N 0.000 description 8
- 239000012634 fragment Substances 0.000 description 8
- 238000011282 treatment Methods 0.000 description 8
- 238000000137 annealing Methods 0.000 description 7
- 238000012217 deletion Methods 0.000 description 7
- 230000037430 deletion Effects 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 6
- 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 6
- 108091028043 Nucleic acid sequence Proteins 0.000 description 6
- 230000037433 frameshift Effects 0.000 description 6
- 230000002068 genetic effect Effects 0.000 description 6
- 108010025306 histidylleucine Proteins 0.000 description 6
- 238000003780 insertion Methods 0.000 description 6
- 230000037431 insertion Effects 0.000 description 6
- HVHRPWQEQHIQJF-AVGNSLFASA-N Leu-Lys-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O HVHRPWQEQHIQJF-AVGNSLFASA-N 0.000 description 5
- 238000012408 PCR amplification Methods 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 239000013604 expression vector Substances 0.000 description 5
- 238000006467 substitution reaction Methods 0.000 description 5
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 4
- 108091033409 CRISPR Proteins 0.000 description 4
- ZOXBSICWUDAOHX-GUBZILKMSA-N Glu-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCC(O)=O ZOXBSICWUDAOHX-GUBZILKMSA-N 0.000 description 4
- BCYGDJXHAGZNPQ-DCAQKATOSA-N Glu-Lys-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O BCYGDJXHAGZNPQ-DCAQKATOSA-N 0.000 description 4
- KFKWRHQBZQICHA-STQMWFEESA-N L-leucyl-L-phenylalanine Natural products CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KFKWRHQBZQICHA-STQMWFEESA-N 0.000 description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 4
- 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 4
- IAJOBQBIJHVGMQ-UHFFFAOYSA-N Phosphinothricin Natural products CP(O)(=O)CCC(N)C(O)=O IAJOBQBIJHVGMQ-UHFFFAOYSA-N 0.000 description 4
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 4
- 108010038633 aspartylglutamate Proteins 0.000 description 4
- 238000010367 cloning Methods 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 4
- 108010049041 glutamylalanine Proteins 0.000 description 4
- 108010050848 glycylleucine Proteins 0.000 description 4
- 108010034529 leucyl-lysine Proteins 0.000 description 4
- 108010044056 leucyl-phenylalanine Proteins 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 230000001404 mediated effect Effects 0.000 description 4
- 108010090894 prolylleucine Proteins 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 108010061238 threonyl-glycine Proteins 0.000 description 4
- 230000009466 transformation Effects 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- 108700026244 Open Reading Frames Proteins 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 238000000227 grinding Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000021121 meiosis Effects 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- SCAKQYSGEIHPLV-IUCAKERBSA-N (4S)-4-[(2-aminoacetyl)amino]-5-[(2S)-2-(carboxymethylcarbamoyl)pyrrolidin-1-yl]-5-oxopentanoic acid Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O SCAKQYSGEIHPLV-IUCAKERBSA-N 0.000 description 2
- 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 2
- IMMKUCQIKKXKNP-DCAQKATOSA-N Ala-Arg-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](C)N)CCCN=C(N)N IMMKUCQIKKXKNP-DCAQKATOSA-N 0.000 description 2
- LGFCAXJBAZESCF-ACZMJKKPSA-N Ala-Gln-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O LGFCAXJBAZESCF-ACZMJKKPSA-N 0.000 description 2
- 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 2
- BVSGPHDECMJBDE-HGNGGELXSA-N Ala-Glu-His Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N BVSGPHDECMJBDE-HGNGGELXSA-N 0.000 description 2
- HXNNRBHASOSVPG-GUBZILKMSA-N Ala-Glu-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O HXNNRBHASOSVPG-GUBZILKMSA-N 0.000 description 2
- 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 2
- WGDNWOMKBUXFHR-BQBZGAKWSA-N Ala-Gly-Arg Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N WGDNWOMKBUXFHR-BQBZGAKWSA-N 0.000 description 2
- SMCGQGDVTPFXKB-XPUUQOCRSA-N Ala-Gly-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N SMCGQGDVTPFXKB-XPUUQOCRSA-N 0.000 description 2
- GRPHQEMIFDPKOE-HGNGGELXSA-N Ala-His-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(O)=O GRPHQEMIFDPKOE-HGNGGELXSA-N 0.000 description 2
- LXAARTARZJJCMB-CIQUZCHMSA-N Ala-Ile-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LXAARTARZJJCMB-CIQUZCHMSA-N 0.000 description 2
- LNNSWWRRYJLGNI-NAKRPEOUSA-N Ala-Ile-Val Chemical compound C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O LNNSWWRRYJLGNI-NAKRPEOUSA-N 0.000 description 2
- LBYMZCVBOKYZNS-CIUDSAMLSA-N Ala-Leu-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O LBYMZCVBOKYZNS-CIUDSAMLSA-N 0.000 description 2
- RTZCUEHYUQZIDE-WHFBIAKZSA-N Ala-Ser-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RTZCUEHYUQZIDE-WHFBIAKZSA-N 0.000 description 2
- MMLHRUJLOUSRJX-CIUDSAMLSA-N Ala-Ser-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN MMLHRUJLOUSRJX-CIUDSAMLSA-N 0.000 description 2
- WQKAQKZRDIZYNV-VZFHVOOUSA-N Ala-Ser-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WQKAQKZRDIZYNV-VZFHVOOUSA-N 0.000 description 2
- GXCSUJQOECMKPV-CIUDSAMLSA-N Arg-Ala-Gln Chemical compound C[C@H](NC(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O GXCSUJQOECMKPV-CIUDSAMLSA-N 0.000 description 2
- UXJCMQFPDWCHKX-DCAQKATOSA-N Arg-Arg-Glu Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O UXJCMQFPDWCHKX-DCAQKATOSA-N 0.000 description 2
- OVVUNXXROOFSIM-SDDRHHMPSA-N Arg-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O OVVUNXXROOFSIM-SDDRHHMPSA-N 0.000 description 2
- NABSCJGZKWSNHX-RCWTZXSCSA-N Arg-Arg-Thr Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H]([C@H](O)C)C(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N NABSCJGZKWSNHX-RCWTZXSCSA-N 0.000 description 2
- SQKPKIJVWHAWNF-DCAQKATOSA-N Arg-Asp-Lys Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(O)=O SQKPKIJVWHAWNF-DCAQKATOSA-N 0.000 description 2
- YNSGXDWWPCGGQS-YUMQZZPRSA-N Arg-Gly-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(O)=O YNSGXDWWPCGGQS-YUMQZZPRSA-N 0.000 description 2
- YKBHOXLMMPZPHQ-GMOBBJLQSA-N Arg-Ile-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O YKBHOXLMMPZPHQ-GMOBBJLQSA-N 0.000 description 2
- OKKMBOSPBDASEP-CYDGBPFRSA-N Arg-Ile-Met Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCSC)C(O)=O OKKMBOSPBDASEP-CYDGBPFRSA-N 0.000 description 2
- DNUKXVMPARLPFN-XUXIUFHCSA-N Arg-Leu-Ile Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O DNUKXVMPARLPFN-XUXIUFHCSA-N 0.000 description 2
- FSNVAJOPUDVQAR-AVGNSLFASA-N Arg-Lys-Arg Chemical compound NC(=N)NCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FSNVAJOPUDVQAR-AVGNSLFASA-N 0.000 description 2
- 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
- XSPKAHFVDKRGRL-DCAQKATOSA-N Arg-Pro-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O XSPKAHFVDKRGRL-DCAQKATOSA-N 0.000 description 2
- KEZVOBAKAXHMOF-GUBZILKMSA-N Arg-Val-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCN=C(N)N KEZVOBAKAXHMOF-GUBZILKMSA-N 0.000 description 2
- ISVACHFCVRKIDG-SRVKXCTJSA-N Arg-Val-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O ISVACHFCVRKIDG-SRVKXCTJSA-N 0.000 description 2
- LLQIAIUAKGNOSE-NHCYSSNCSA-N Arg-Val-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCN=C(N)N LLQIAIUAKGNOSE-NHCYSSNCSA-N 0.000 description 2
- 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 2
- WIDVAWAQBRAKTI-YUMQZZPRSA-N Asn-Leu-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O WIDVAWAQBRAKTI-YUMQZZPRSA-N 0.000 description 2
- KRXIWXCXOARFNT-ZLUOBGJFSA-N Asp-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O KRXIWXCXOARFNT-ZLUOBGJFSA-N 0.000 description 2
- HOQGTAIGQSDCHR-SRVKXCTJSA-N Asp-Asn-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HOQGTAIGQSDCHR-SRVKXCTJSA-N 0.000 description 2
- SBHUBSDEZQFJHJ-CIUDSAMLSA-N Asp-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(O)=O SBHUBSDEZQFJHJ-CIUDSAMLSA-N 0.000 description 2
- HRGGPWBIMIQANI-GUBZILKMSA-N Asp-Gln-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O HRGGPWBIMIQANI-GUBZILKMSA-N 0.000 description 2
- 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 2
- YDJVIBMKAMQPPP-LAEOZQHASA-N Asp-Glu-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O YDJVIBMKAMQPPP-LAEOZQHASA-N 0.000 description 2
- YNCHFVRXEQFPBY-BQBZGAKWSA-N Asp-Gly-Arg Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N YNCHFVRXEQFPBY-BQBZGAKWSA-N 0.000 description 2
- 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 2
- CJUKAWUWBZCTDQ-SRVKXCTJSA-N Asp-Leu-Lys Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O CJUKAWUWBZCTDQ-SRVKXCTJSA-N 0.000 description 2
- LIVXPXUVXFRWNY-CIUDSAMLSA-N Asp-Lys-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O LIVXPXUVXFRWNY-CIUDSAMLSA-N 0.000 description 2
- VSMYBNPOHYAXSD-GUBZILKMSA-N Asp-Lys-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O VSMYBNPOHYAXSD-GUBZILKMSA-N 0.000 description 2
- ZVGRHIRJLWBWGJ-ACZMJKKPSA-N Asp-Ser-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZVGRHIRJLWBWGJ-ACZMJKKPSA-N 0.000 description 2
- NBKLEMWHDLAUEM-CIUDSAMLSA-N Asp-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)O)N NBKLEMWHDLAUEM-CIUDSAMLSA-N 0.000 description 2
- PLBJMUUEGBBHRH-ZLUOBGJFSA-N Cys-Ala-Asn Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O PLBJMUUEGBBHRH-ZLUOBGJFSA-N 0.000 description 2
- KFYPRIGJTICABD-XGEHTFHBSA-N Cys-Thr-Val Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CS)N)O KFYPRIGJTICABD-XGEHTFHBSA-N 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- KVYVOGYEMPEXBT-GUBZILKMSA-N Gln-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O KVYVOGYEMPEXBT-GUBZILKMSA-N 0.000 description 2
- 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 2
- MCAVASRGVBVPMX-FXQIFTODSA-N Gln-Glu-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O MCAVASRGVBVPMX-FXQIFTODSA-N 0.000 description 2
- IKFZXRLDMYWNBU-YUMQZZPRSA-N Gln-Gly-Arg Chemical compound NC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N IKFZXRLDMYWNBU-YUMQZZPRSA-N 0.000 description 2
- MLSKFHLRFVGNLL-WDCWCFNPSA-N Gln-Leu-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MLSKFHLRFVGNLL-WDCWCFNPSA-N 0.000 description 2
- ZXGLLNZQSBLQLT-SRVKXCTJSA-N Gln-Met-Lys Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)N)N ZXGLLNZQSBLQLT-SRVKXCTJSA-N 0.000 description 2
- HMIXCETWRYDVMO-GUBZILKMSA-N Gln-Pro-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O HMIXCETWRYDVMO-GUBZILKMSA-N 0.000 description 2
- UTKUTMJSWKKHEM-WDSKDSINSA-N Glu-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(O)=O UTKUTMJSWKKHEM-WDSKDSINSA-N 0.000 description 2
- WOMUDRVDJMHTCV-DCAQKATOSA-N Glu-Arg-Arg Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WOMUDRVDJMHTCV-DCAQKATOSA-N 0.000 description 2
- KKCUFHUTMKQQCF-SRVKXCTJSA-N Glu-Arg-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O KKCUFHUTMKQQCF-SRVKXCTJSA-N 0.000 description 2
- DYFJZDDQPNIPAB-NHCYSSNCSA-N Glu-Arg-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(O)=O DYFJZDDQPNIPAB-NHCYSSNCSA-N 0.000 description 2
- DSPQRJXOIXHOHK-WDSKDSINSA-N Glu-Asp-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O DSPQRJXOIXHOHK-WDSKDSINSA-N 0.000 description 2
- CJWANNXUTOATSJ-DCAQKATOSA-N Glu-Gln-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)O)N CJWANNXUTOATSJ-DCAQKATOSA-N 0.000 description 2
- PVBBEKPHARMPHX-DCAQKATOSA-N Glu-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCC(O)=O PVBBEKPHARMPHX-DCAQKATOSA-N 0.000 description 2
- HTTSBEBKVNEDFE-AUTRQRHGSA-N Glu-Gln-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)O)N HTTSBEBKVNEDFE-AUTRQRHGSA-N 0.000 description 2
- NKLRYVLERDYDBI-FXQIFTODSA-N Glu-Glu-Asp Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O NKLRYVLERDYDBI-FXQIFTODSA-N 0.000 description 2
- AUTNXSQEVVHSJK-YVNDNENWSA-N Glu-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)CCC(O)=O AUTNXSQEVVHSJK-YVNDNENWSA-N 0.000 description 2
- OAGVHWYIBZMWLA-YFKPBYRVSA-N Glu-Gly-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)NCC(O)=O OAGVHWYIBZMWLA-YFKPBYRVSA-N 0.000 description 2
- KRGZZKWSBGPLKL-IUCAKERBSA-N Glu-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)O)N KRGZZKWSBGPLKL-IUCAKERBSA-N 0.000 description 2
- QIQABBIDHGQXGA-ZPFDUUQYSA-N Glu-Ile-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O QIQABBIDHGQXGA-ZPFDUUQYSA-N 0.000 description 2
- LGYCLOCORAEQSZ-PEFMBERDSA-N Glu-Ile-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O LGYCLOCORAEQSZ-PEFMBERDSA-N 0.000 description 2
- 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 2
- HVYWQYLBVXMXSV-GUBZILKMSA-N Glu-Leu-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O HVYWQYLBVXMXSV-GUBZILKMSA-N 0.000 description 2
- DNPCBMNFQVTHMA-DCAQKATOSA-N Glu-Leu-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O DNPCBMNFQVTHMA-DCAQKATOSA-N 0.000 description 2
- OQXDUSZKISQQSS-GUBZILKMSA-N Glu-Lys-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O OQXDUSZKISQQSS-GUBZILKMSA-N 0.000 description 2
- JDUKCSSHWNIQQZ-IHRRRGAJSA-N Glu-Phe-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JDUKCSSHWNIQQZ-IHRRRGAJSA-N 0.000 description 2
- BIYNPVYAZOUVFQ-CIUDSAMLSA-N Glu-Pro-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O BIYNPVYAZOUVFQ-CIUDSAMLSA-N 0.000 description 2
- GPSHCSTUYOQPAI-JHEQGTHGSA-N Glu-Thr-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O GPSHCSTUYOQPAI-JHEQGTHGSA-N 0.000 description 2
- LZEUDRYSAZAJIO-AUTRQRHGSA-N Glu-Val-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LZEUDRYSAZAJIO-AUTRQRHGSA-N 0.000 description 2
- 239000005561 Glufosinate Substances 0.000 description 2
- PYTZFYUXZZHOAD-WHFBIAKZSA-N Gly-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)CN PYTZFYUXZZHOAD-WHFBIAKZSA-N 0.000 description 2
- QXPRJQPCFXMCIY-NKWVEPMBSA-N Gly-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN QXPRJQPCFXMCIY-NKWVEPMBSA-N 0.000 description 2
- LJPIRKICOISLKN-WHFBIAKZSA-N Gly-Ala-Ser Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O LJPIRKICOISLKN-WHFBIAKZSA-N 0.000 description 2
- UPOJUWHGMDJUQZ-IUCAKERBSA-N Gly-Arg-Arg Chemical compound NC(=N)NCCC[C@H](NC(=O)CN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O UPOJUWHGMDJUQZ-IUCAKERBSA-N 0.000 description 2
- CLODWIOAKCSBAN-BQBZGAKWSA-N Gly-Arg-Asp Chemical compound NC(N)=NCCC[C@H](NC(=O)CN)C(=O)N[C@@H](CC(O)=O)C(O)=O CLODWIOAKCSBAN-BQBZGAKWSA-N 0.000 description 2
- KKBWDNZXYLGJEY-UHFFFAOYSA-N Gly-Arg-Pro Natural products NCC(=O)NC(CCNC(=N)N)C(=O)N1CCCC1C(=O)O KKBWDNZXYLGJEY-UHFFFAOYSA-N 0.000 description 2
- LCNXZQROPKFGQK-WHFBIAKZSA-N Gly-Asp-Ser Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O LCNXZQROPKFGQK-WHFBIAKZSA-N 0.000 description 2
- 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 2
- UFPXDFOYHVEIPI-BYPYZUCNSA-N Gly-Gly-Asp Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC(O)=O UFPXDFOYHVEIPI-BYPYZUCNSA-N 0.000 description 2
- GDOZQTNZPCUARW-YFKPBYRVSA-N Gly-Gly-Glu Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O GDOZQTNZPCUARW-YFKPBYRVSA-N 0.000 description 2
- ORXZVPZCPMKHNR-IUCAKERBSA-N Gly-His-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CNC=N1 ORXZVPZCPMKHNR-IUCAKERBSA-N 0.000 description 2
- CQIIXEHDSZUSAG-QWRGUYRKSA-N Gly-His-His Chemical compound C([C@H](NC(=O)CN)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 CQIIXEHDSZUSAG-QWRGUYRKSA-N 0.000 description 2
- FXGRXIATVXUAHO-WEDXCCLWSA-N Gly-Lys-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCCN FXGRXIATVXUAHO-WEDXCCLWSA-N 0.000 description 2
- ZWRDOVYMQAAISL-UWVGGRQHSA-N Gly-Met-Lys Chemical compound CSCC[C@H](NC(=O)CN)C(=O)N[C@H](C(O)=O)CCCCN ZWRDOVYMQAAISL-UWVGGRQHSA-N 0.000 description 2
- QIVPRLJQQVXCIY-HGNGGELXSA-N His-Ala-Gln Chemical compound C[C@H](NC(=O)[C@@H](N)Cc1cnc[nH]1)C(=O)N[C@@H](CCC(N)=O)C(O)=O QIVPRLJQQVXCIY-HGNGGELXSA-N 0.000 description 2
- 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 2
- KFQDSSNYWKZFOO-LSJOCFKGSA-N His-Val-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O KFQDSSNYWKZFOO-LSJOCFKGSA-N 0.000 description 2
- JRHFQUPIZOYKQP-KBIXCLLPSA-N Ile-Ala-Glu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(O)=O JRHFQUPIZOYKQP-KBIXCLLPSA-N 0.000 description 2
- MKWSZEHGHSLNPF-NAKRPEOUSA-N Ile-Ala-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)O)N MKWSZEHGHSLNPF-NAKRPEOUSA-N 0.000 description 2
- RGSOCXHDOPQREB-ZPFDUUQYSA-N Ile-Asp-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N RGSOCXHDOPQREB-ZPFDUUQYSA-N 0.000 description 2
- CCHSQWLCOOZREA-GMOBBJLQSA-N Ile-Asp-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCSC)C(=O)O)N CCHSQWLCOOZREA-GMOBBJLQSA-N 0.000 description 2
- KUHFPGIVBOCRMV-MNXVOIDGSA-N Ile-Gln-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(C)C)C(=O)O)N KUHFPGIVBOCRMV-MNXVOIDGSA-N 0.000 description 2
- DVRDRICMWUSCBN-UKJIMTQDSA-N Ile-Gln-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N DVRDRICMWUSCBN-UKJIMTQDSA-N 0.000 description 2
- NZOCIWKZUVUNDW-ZKWXMUAHSA-N Ile-Gly-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O NZOCIWKZUVUNDW-ZKWXMUAHSA-N 0.000 description 2
- TWPSALMCEHCIOY-YTFOTSKYSA-N Ile-Ile-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(=O)O)N TWPSALMCEHCIOY-YTFOTSKYSA-N 0.000 description 2
- IOVUXUSIGXCREV-DKIMLUQUSA-N Ile-Leu-Phe Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 IOVUXUSIGXCREV-DKIMLUQUSA-N 0.000 description 2
- GVKKVHNRTUFCCE-BJDJZHNGSA-N Ile-Leu-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)O)N GVKKVHNRTUFCCE-BJDJZHNGSA-N 0.000 description 2
- BKPPWVSPSIUXHZ-OSUNSFLBSA-N Ile-Met-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N BKPPWVSPSIUXHZ-OSUNSFLBSA-N 0.000 description 2
- MLSUZXHSNRBDCI-CYDGBPFRSA-N Ile-Pro-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)O)N MLSUZXHSNRBDCI-CYDGBPFRSA-N 0.000 description 2
- JZNVOBUNTWNZPW-GHCJXIJMSA-N Ile-Ser-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N JZNVOBUNTWNZPW-GHCJXIJMSA-N 0.000 description 2
- VGSPNSSCMOHRRR-BJDJZHNGSA-N Ile-Ser-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N VGSPNSSCMOHRRR-BJDJZHNGSA-N 0.000 description 2
- DLEBSGAVWRPTIX-PEDHHIEDSA-N Ile-Val-Ile Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)[C@@H](C)CC DLEBSGAVWRPTIX-PEDHHIEDSA-N 0.000 description 2
- 108010065920 Insulin Lispro Proteins 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- 241000880493 Leptailurus serval Species 0.000 description 2
- MJOZZTKJZQFKDK-GUBZILKMSA-N Leu-Ala-Gln Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(N)=O MJOZZTKJZQFKDK-GUBZILKMSA-N 0.000 description 2
- 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 2
- CNNQBZRGQATKNY-DCAQKATOSA-N Leu-Arg-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CS)C(=O)O)N CNNQBZRGQATKNY-DCAQKATOSA-N 0.000 description 2
- 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 2
- DBVWMYGBVFCRBE-CIUDSAMLSA-N Leu-Asn-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O DBVWMYGBVFCRBE-CIUDSAMLSA-N 0.000 description 2
- ILJREDZFPHTUIE-GUBZILKMSA-N Leu-Asp-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ILJREDZFPHTUIE-GUBZILKMSA-N 0.000 description 2
- 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 2
- ZTLGVASZOIKNIX-DCAQKATOSA-N Leu-Gln-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ZTLGVASZOIKNIX-DCAQKATOSA-N 0.000 description 2
- CQGSYZCULZMEDE-SRVKXCTJSA-N Leu-Gln-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(O)=O CQGSYZCULZMEDE-SRVKXCTJSA-N 0.000 description 2
- CQGSYZCULZMEDE-UHFFFAOYSA-N Leu-Gln-Pro Natural products CC(C)CC(N)C(=O)NC(CCC(N)=O)C(=O)N1CCCC1C(O)=O CQGSYZCULZMEDE-UHFFFAOYSA-N 0.000 description 2
- 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 2
- BABSVXFGKFLIGW-UWVGGRQHSA-N Leu-Gly-Arg Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCNC(N)=N BABSVXFGKFLIGW-UWVGGRQHSA-N 0.000 description 2
- PBGDOSARRIJMEV-DLOVCJGASA-N Leu-His-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(O)=O PBGDOSARRIJMEV-DLOVCJGASA-N 0.000 description 2
- AUBMZAMQCOYSIC-MNXVOIDGSA-N Leu-Ile-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(O)=O AUBMZAMQCOYSIC-MNXVOIDGSA-N 0.000 description 2
- HGFGEMSVBMCFKK-MNXVOIDGSA-N Leu-Ile-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O HGFGEMSVBMCFKK-MNXVOIDGSA-N 0.000 description 2
- 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 2
- 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 2
- JNDYEOUZBLOVOF-AVGNSLFASA-N Leu-Leu-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O JNDYEOUZBLOVOF-AVGNSLFASA-N 0.000 description 2
- QNBVTHNJGCOVFA-AVGNSLFASA-N Leu-Leu-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCC(O)=O QNBVTHNJGCOVFA-AVGNSLFASA-N 0.000 description 2
- YOKVEHGYYQEQOP-QWRGUYRKSA-N Leu-Leu-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YOKVEHGYYQEQOP-QWRGUYRKSA-N 0.000 description 2
- PJWOOBTYQNNRBF-BZSNNMDCSA-N Leu-Phe-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)O)N PJWOOBTYQNNRBF-BZSNNMDCSA-N 0.000 description 2
- ADJWHHZETYAAAX-SRVKXCTJSA-N Leu-Ser-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N ADJWHHZETYAAAX-SRVKXCTJSA-N 0.000 description 2
- ICYRCNICGBJLGM-HJGDQZAQSA-N Leu-Thr-Asp 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(O)=O ICYRCNICGBJLGM-HJGDQZAQSA-N 0.000 description 2
- LFSQWRSVPNKJGP-WDCWCFNPSA-N Leu-Thr-Glu 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(O)=O LFSQWRSVPNKJGP-WDCWCFNPSA-N 0.000 description 2
- URHJPNHRQMQGOZ-RHYQMDGZSA-N Leu-Thr-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(O)=O URHJPNHRQMQGOZ-RHYQMDGZSA-N 0.000 description 2
- KLSUAWUZBMAZCL-RHYQMDGZSA-N Leu-Thr-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(O)=O KLSUAWUZBMAZCL-RHYQMDGZSA-N 0.000 description 2
- FBNPMTNBFFAMMH-AVGNSLFASA-N Leu-Val-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N FBNPMTNBFFAMMH-AVGNSLFASA-N 0.000 description 2
- FBNPMTNBFFAMMH-UHFFFAOYSA-N Leu-Val-Arg Natural products CC(C)CC(N)C(=O)NC(C(C)C)C(=O)NC(C(O)=O)CCCN=C(N)N FBNPMTNBFFAMMH-UHFFFAOYSA-N 0.000 description 2
- XZNJZXJZBMBGGS-NHCYSSNCSA-N Leu-Val-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O XZNJZXJZBMBGGS-NHCYSSNCSA-N 0.000 description 2
- AIMGJYMCTAABEN-GVXVVHGQSA-N Leu-Val-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIMGJYMCTAABEN-GVXVVHGQSA-N 0.000 description 2
- 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 2
- 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 2
- ABHIXYDMILIUKV-CIUDSAMLSA-N Lys-Asn-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O ABHIXYDMILIUKV-CIUDSAMLSA-N 0.000 description 2
- DGWXCIORNLWGGG-CIUDSAMLSA-N Lys-Asn-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O DGWXCIORNLWGGG-CIUDSAMLSA-N 0.000 description 2
- LMVOVCYVZBBWQB-SRVKXCTJSA-N Lys-Asp-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN LMVOVCYVZBBWQB-SRVKXCTJSA-N 0.000 description 2
- 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 2
- OVAOHZIOUBEQCJ-IHRRRGAJSA-N Lys-Leu-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O OVAOHZIOUBEQCJ-IHRRRGAJSA-N 0.000 description 2
- RBEATVHTWHTHTJ-KKUMJFAQSA-N Lys-Leu-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O RBEATVHTWHTHTJ-KKUMJFAQSA-N 0.000 description 2
- YPLVCBKEPJPBDQ-MELADBBJSA-N Lys-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCCN)N YPLVCBKEPJPBDQ-MELADBBJSA-N 0.000 description 2
- ZJWIXBZTAAJERF-IHRRRGAJSA-N Lys-Lys-Arg Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CCCN=C(N)N ZJWIXBZTAAJERF-IHRRRGAJSA-N 0.000 description 2
- KJIXWRWPOCKYLD-IHRRRGAJSA-N Lys-Lys-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)N KJIXWRWPOCKYLD-IHRRRGAJSA-N 0.000 description 2
- YFQSSOAGMZGXFT-MEYUZBJRSA-N Lys-Thr-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O YFQSSOAGMZGXFT-MEYUZBJRSA-N 0.000 description 2
- XYLSGAWRCZECIQ-JYJNAYRXSA-N Lys-Tyr-Glu Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCC(O)=O)C(O)=O)CC1=CC=C(O)C=C1 XYLSGAWRCZECIQ-JYJNAYRXSA-N 0.000 description 2
- IHITVQKJXQQGLJ-LPEHRKFASA-N Met-Asn-Pro Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N IHITVQKJXQQGLJ-LPEHRKFASA-N 0.000 description 2
- GODBLDDYHFTUAH-CIUDSAMLSA-N Met-Asp-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCC(O)=O GODBLDDYHFTUAH-CIUDSAMLSA-N 0.000 description 2
- FYRUJIJAUPHUNB-IUCAKERBSA-N Met-Gly-Arg Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCNC(N)=N FYRUJIJAUPHUNB-IUCAKERBSA-N 0.000 description 2
- AFFKUNVPPLQUGA-DCAQKATOSA-N Met-Leu-Ala Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O AFFKUNVPPLQUGA-DCAQKATOSA-N 0.000 description 2
- CQRGINSEMFBACV-WPRPVWTQSA-N Met-Val-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O CQRGINSEMFBACV-WPRPVWTQSA-N 0.000 description 2
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 2
- 108010065395 Neuropep-1 Proteins 0.000 description 2
- OFBQJSOFQDEBGM-UHFFFAOYSA-N Pentane Chemical compound CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 description 2
- 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 2
- HTKNPQZCMLBOTQ-XVSYOHENSA-N Phe-Asn-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CC=CC=C1)N)O HTKNPQZCMLBOTQ-XVSYOHENSA-N 0.000 description 2
- ZENDEDYRYVHBEG-SRVKXCTJSA-N Phe-Asp-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 ZENDEDYRYVHBEG-SRVKXCTJSA-N 0.000 description 2
- MQVFHOPCKNTHGT-MELADBBJSA-N Phe-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O MQVFHOPCKNTHGT-MELADBBJSA-N 0.000 description 2
- GYEPCBNTTRORKW-PCBIJLKTSA-N Phe-Ile-Asp Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O GYEPCBNTTRORKW-PCBIJLKTSA-N 0.000 description 2
- SMFGCTXUBWEPKM-KBPBESRZSA-N Phe-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 SMFGCTXUBWEPKM-KBPBESRZSA-N 0.000 description 2
- KLYYKKGCPOGDPE-OEAJRASXSA-N Phe-Thr-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O KLYYKKGCPOGDPE-OEAJRASXSA-N 0.000 description 2
- CDHURCQGUDNBMA-UBHSHLNASA-N Phe-Val-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 CDHURCQGUDNBMA-UBHSHLNASA-N 0.000 description 2
- FISHYTLIMUYTQY-GUBZILKMSA-N Pro-Gln-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1 FISHYTLIMUYTQY-GUBZILKMSA-N 0.000 description 2
- VDGTVWFMRXVQCT-GUBZILKMSA-N Pro-Glu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CCCN1 VDGTVWFMRXVQCT-GUBZILKMSA-N 0.000 description 2
- QEWBZBLXDKIQPS-STQMWFEESA-N Pro-Gly-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O QEWBZBLXDKIQPS-STQMWFEESA-N 0.000 description 2
- FHZJRBVMLGOHBX-GUBZILKMSA-N Pro-Pro-Asp Chemical compound OC(=O)C[C@H](NC(=O)[C@@H]1CCCN1C(=O)[C@@H]1CCCN1)C(O)=O FHZJRBVMLGOHBX-GUBZILKMSA-N 0.000 description 2
- 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 2
- RMJZWERKFFNNNS-XGEHTFHBSA-N Pro-Thr-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O RMJZWERKFFNNNS-XGEHTFHBSA-N 0.000 description 2
- 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 2
- 108020004511 Recombinant DNA Proteins 0.000 description 2
- ZUGXSSFMTXKHJS-ZLUOBGJFSA-N Ser-Ala-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O ZUGXSSFMTXKHJS-ZLUOBGJFSA-N 0.000 description 2
- MMGJPDWSIOAGTH-ACZMJKKPSA-N Ser-Ala-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MMGJPDWSIOAGTH-ACZMJKKPSA-N 0.000 description 2
- KNZQGAUEYZJUSQ-ZLUOBGJFSA-N Ser-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CO)N KNZQGAUEYZJUSQ-ZLUOBGJFSA-N 0.000 description 2
- QPFJSHSJFIYDJZ-GHCJXIJMSA-N Ser-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CO QPFJSHSJFIYDJZ-GHCJXIJMSA-N 0.000 description 2
- FMDHKPRACUXATF-ACZMJKKPSA-N Ser-Gln-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O FMDHKPRACUXATF-ACZMJKKPSA-N 0.000 description 2
- 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 2
- YMTLKLXDFCSCNX-BYPYZUCNSA-N Ser-Gly-Gly Chemical compound OC[C@H](N)C(=O)NCC(=O)NCC(O)=O YMTLKLXDFCSCNX-BYPYZUCNSA-N 0.000 description 2
- UBRMZSHOOIVJPW-SRVKXCTJSA-N Ser-Leu-Lys Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O UBRMZSHOOIVJPW-SRVKXCTJSA-N 0.000 description 2
- YUJLIIRMIAGMCQ-CIUDSAMLSA-N Ser-Leu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YUJLIIRMIAGMCQ-CIUDSAMLSA-N 0.000 description 2
- OCWWJBZQXGYQCA-DCAQKATOSA-N Ser-Lys-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(O)=O OCWWJBZQXGYQCA-DCAQKATOSA-N 0.000 description 2
- KJKQUQXDEKMPDK-FXQIFTODSA-N Ser-Met-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(O)=O KJKQUQXDEKMPDK-FXQIFTODSA-N 0.000 description 2
- 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 2
- 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 2
- PMTWIUBUQRGCSB-FXQIFTODSA-N Ser-Val-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O PMTWIUBUQRGCSB-FXQIFTODSA-N 0.000 description 2
- ANOQEBQWIAYIMV-AEJSXWLSSA-N Ser-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N ANOQEBQWIAYIMV-AEJSXWLSSA-N 0.000 description 2
- XSLXHSYIVPGEER-KZVJFYERSA-N Thr-Ala-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O XSLXHSYIVPGEER-KZVJFYERSA-N 0.000 description 2
- VFEHSAJCWWHDBH-RHYQMDGZSA-N Thr-Arg-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O VFEHSAJCWWHDBH-RHYQMDGZSA-N 0.000 description 2
- QGXCWPNQVCYJEL-NUMRIWBASA-N Thr-Asn-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O QGXCWPNQVCYJEL-NUMRIWBASA-N 0.000 description 2
- JBHMLZSKIXMVFS-XVSYOHENSA-N Thr-Asn-Phe Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JBHMLZSKIXMVFS-XVSYOHENSA-N 0.000 description 2
- SHOMROOOQBDGRL-JHEQGTHGSA-N Thr-Glu-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O SHOMROOOQBDGRL-JHEQGTHGSA-N 0.000 description 2
- JKGGPMOUIAAJAA-YEPSODPASA-N Thr-Gly-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O JKGGPMOUIAAJAA-YEPSODPASA-N 0.000 description 2
- FKIGTIXHSRNKJU-IXOXFDKPSA-N Thr-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@H](O)C)CC1=CN=CN1 FKIGTIXHSRNKJU-IXOXFDKPSA-N 0.000 description 2
- 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 2
- FYBFTPLPAXZBOY-KKHAAJSZSA-N Thr-Val-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O FYBFTPLPAXZBOY-KKHAAJSZSA-N 0.000 description 2
- AKHDFZHUPGVFEJ-YEPSODPASA-N Thr-Val-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O AKHDFZHUPGVFEJ-YEPSODPASA-N 0.000 description 2
- 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 2
- MNYNCKZAEIAONY-XGEHTFHBSA-N Thr-Val-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O MNYNCKZAEIAONY-XGEHTFHBSA-N 0.000 description 2
- SEXRBCGSZRCIPE-LYSGOOTNSA-N Trp-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N)O SEXRBCGSZRCIPE-LYSGOOTNSA-N 0.000 description 2
- GFZQWWDXJVGEMW-ULQDDVLXSA-N Tyr-Arg-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N)O GFZQWWDXJVGEMW-ULQDDVLXSA-N 0.000 description 2
- NRFTYDWKWGJLAR-MELADBBJSA-N Tyr-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)C(=O)O NRFTYDWKWGJLAR-MELADBBJSA-N 0.000 description 2
- CNLKDWSAORJEMW-KWQFWETISA-N Tyr-Gly-Ala Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H](C)C(O)=O CNLKDWSAORJEMW-KWQFWETISA-N 0.000 description 2
- TYFLVOUZHQUBGM-IHRRRGAJSA-N Tyr-Ser-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 TYFLVOUZHQUBGM-IHRRRGAJSA-N 0.000 description 2
- VLOYGOZDPGYWFO-LAEOZQHASA-N Val-Asp-Glu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O VLOYGOZDPGYWFO-LAEOZQHASA-N 0.000 description 2
- SZTTYWIUCGSURQ-AUTRQRHGSA-N Val-Glu-Glu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O SZTTYWIUCGSURQ-AUTRQRHGSA-N 0.000 description 2
- 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 2
- OQWNEUXPKHIEJO-NRPADANISA-N Val-Glu-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N OQWNEUXPKHIEJO-NRPADANISA-N 0.000 description 2
- SYOMXKPPFZRELL-ONGXEEELSA-N Val-Gly-Lys Chemical compound CC(C)[C@@H](C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)O)N SYOMXKPPFZRELL-ONGXEEELSA-N 0.000 description 2
- KDKLLPMFFGYQJD-CYDGBPFRSA-N Val-Ile-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](C(C)C)N KDKLLPMFFGYQJD-CYDGBPFRSA-N 0.000 description 2
- FTKXYXACXYOHND-XUXIUFHCSA-N Val-Ile-Leu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O FTKXYXACXYOHND-XUXIUFHCSA-N 0.000 description 2
- AGXGCFSECFQMKB-NHCYSSNCSA-N Val-Leu-Asp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N AGXGCFSECFQMKB-NHCYSSNCSA-N 0.000 description 2
- UMPVMAYCLYMYGA-ONGXEEELSA-N Val-Leu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O UMPVMAYCLYMYGA-ONGXEEELSA-N 0.000 description 2
- ZXYPHBKIZLAQTL-QXEWZRGKSA-N Val-Pro-Asp Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(=O)O)C(=O)O)N ZXYPHBKIZLAQTL-QXEWZRGKSA-N 0.000 description 2
- GUIYPEKUEMQBIK-JSGCOSHPSA-N Val-Tyr-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)NCC(O)=O GUIYPEKUEMQBIK-JSGCOSHPSA-N 0.000 description 2
- RTJPAGFXOWEBAI-SRVKXCTJSA-N Val-Val-Arg Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N RTJPAGFXOWEBAI-SRVKXCTJSA-N 0.000 description 2
- 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 2
- 108010041407 alanylaspartic acid Proteins 0.000 description 2
- 108010005233 alanylglutamic acid Proteins 0.000 description 2
- 108010047495 alanylglycine Proteins 0.000 description 2
- 108010070944 alanylhistidine Proteins 0.000 description 2
- 108010089975 arginyl-glycyl-aspartyl-serine Proteins 0.000 description 2
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 2
- 108010062796 arginyllysine Proteins 0.000 description 2
- 108010077245 asparaginyl-proline Proteins 0.000 description 2
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 2
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 2
- 108010047857 aspartylglycine Proteins 0.000 description 2
- 210000004899 c-terminal region Anatomy 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 2
- IAJOBQBIJHVGMQ-BYPYZUCNSA-N glufosinate-P Chemical compound CP(O)(=O)CC[C@H](N)C(O)=O IAJOBQBIJHVGMQ-BYPYZUCNSA-N 0.000 description 2
- 108010042598 glutamyl-aspartyl-glycine Proteins 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 2
- 108010000434 glycyl-alanyl-leucine Proteins 0.000 description 2
- 108010072405 glycyl-aspartyl-glycine Proteins 0.000 description 2
- 108010026364 glycyl-glycyl-leucine Proteins 0.000 description 2
- 108010079413 glycyl-prolyl-glutamic acid Proteins 0.000 description 2
- 108010010147 glycylglutamine Proteins 0.000 description 2
- 108010020688 glycylhistidine Proteins 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- 108010018006 histidylserine Proteins 0.000 description 2
- 230000001788 irregular Effects 0.000 description 2
- 108010047926 leucyl-lysyl-tyrosine Proteins 0.000 description 2
- 108010003700 lysyl aspartic acid Proteins 0.000 description 2
- 108010017391 lysylvaline Proteins 0.000 description 2
- 210000001161 mammalian embryo Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 108010056582 methionylglutamic acid Proteins 0.000 description 2
- 108010087846 prolyl-prolyl-glycine Proteins 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 2
- 108010026333 seryl-proline Proteins 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 108010005652 splenotritin Proteins 0.000 description 2
- 230000035882 stress Effects 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 238000011426 transformation method Methods 0.000 description 2
- 230000009105 vegetative growth Effects 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 229930024421 Adenine Natural products 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- GFYOIYJJMSHLSN-QXEWZRGKSA-N Asp-Val-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O GFYOIYJJMSHLSN-QXEWZRGKSA-N 0.000 description 1
- 206010003694 Atrophy Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 238000010354 CRISPR gene editing Methods 0.000 description 1
- 108700003861 Dominant Genes Proteins 0.000 description 1
- GJHWILMUOANXTG-WPRPVWTQSA-N Gly-Val-Arg Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O GJHWILMUOANXTG-WPRPVWTQSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 101150062550 HKT1 gene Proteins 0.000 description 1
- HEFNNWSXXWATRW-UHFFFAOYSA-N Ibuprofen Chemical compound CC(C)CC1=CC=C(C(C)C(O)=O)C=C1 HEFNNWSXXWATRW-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- KPYAOIVPJKPIOU-KKUMJFAQSA-N Leu-Lys-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O KPYAOIVPJKPIOU-KKUMJFAQSA-N 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241000209504 Poaceae Species 0.000 description 1
- 238000003559 RNA-seq method Methods 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- 235000007244 Zea mays Nutrition 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000037444 atrophy Effects 0.000 description 1
- 101150103518 bar gene Proteins 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000011712 cell development Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000009402 cross-breeding Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 210000005069 ears Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000012215 gene cloning Methods 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 238000010362 genome editing Methods 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 108010036413 histidylglycine Proteins 0.000 description 1
- 101150062015 hyg gene Proteins 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 210000003794 male germ cell Anatomy 0.000 description 1
- 238000001531 micro-dissection Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 210000003824 plant germ cell Anatomy 0.000 description 1
- 230000008119 pollen development Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000002331 protein detection Methods 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 230000008844 regulatory mechanism Effects 0.000 description 1
- 230000033458 reproduction Effects 0.000 description 1
- 238000007480 sanger sequencing Methods 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000014639 sexual reproduction Effects 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- 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
- C12N15/8287—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for fertility modification, e.g. apomixis
-
- 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
- C12N15/8287—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for fertility modification, e.g. apomixis
- C12N15/8289—Male sterility
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
- C07K2319/22—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a Strep-tag
-
- 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
-
- 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
- C07K2319/43—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation containing a FLAG-tag
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
本发明公开了调控玉米花粉育性和花粉耐热性的方法。本发明提供了氨基酸序列如SEQ ID NO:1所示的蛋白质Ms42和氨基酸序列如SEQ ID NO:3所示的蛋白质ms42。实验证明，蛋白质ms42可以提高花粉耐热性和维持玉米雄性可育，蛋白质Ms42可以促进玉米雄性不育。本发明具有重大的应用价值。
Description
技术领域
本发明属于生物技术领域，具体涉及调控玉米花粉育性和花粉耐热性的方法。
背景技术
玉米(Zeamays L.)属禾本科玉蜀黍属，是我国第一大粮食作物，随着中国经济的发展，玉米播种面积和产量连年增长，在我国农业生产和国民经济中占有十分重要的地位。玉米是杂种优势利用的典范，其中杂交育种和制种技术的关键均在于母本的去雄，而雄性不育系是作物杂种优势利用和经济杂交种生产的宝贵选育材料，利用雄性不育系可以有效降低母本去雄的成本。随着现代育种技术的进步，基于显性雄性不育系的制种系统已在玉米中得到开发和应用。因此，优良显性雄性不育系的选择对于玉米高效的杂交制种发展具有重要价值。
全球气候变化，温度的升高会对植物生殖细胞的发育以及繁殖成功率产生极大损害。而减数分裂是有性生殖的基础，对植物的基因组多样性和倍性稳定具有重要意义。虽然过去几十年，通过生物学的方法鉴定发现了一些影响植物减数分裂的关键基因，但是对其发育的调控网络还不清楚，尤其是环境温度对于植物减数分裂过程的影响，因此探索高温胁迫下植物雄性生殖细胞发育的调控机制和遗传基础对于农业经济安全以及培育耐高温品种具有重要意义。
发明内容
本发明的目的是调控玉米花粉育性和花粉耐热性。
本发明首先保护蛋白质Ms42，可为如下a1)或a2)或a3)或a4)：
a1)氨基酸序列是SEQ ID NO:1所示的蛋白质；
a2)在SEQ ID NO:1所示的蛋白质的N端或/和C端连接标签得到的融合蛋白质；
a3)将a1)或a2)所示的蛋白质经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的与玉米雄性不育相关的蛋白质；
a4)与SEQ ID NO:1限定的氨基酸序列具有80％或80％以上同源性，来源于玉米且与雄性育性不育相关的蛋白质。
其中，SEQ ID NO:1由912个氨基酸残基组成。
为了使a1)中的蛋白质便于纯化，可在SEQ ID NO:1所示的蛋白质的氨基末端或羧基末端连接上如表1所示的标签。
表1.标签的序列
上述a3)中的蛋白质，所述一个或几个氨基酸残基的取代和/或缺失和/或添加为不超过10个氨基酸残基的取代和/或缺失和/或添加。
上述a3)中的蛋白质可人工合成，也可先合成其编码基因，再进行生物表达得到。
上述a3)中的蛋白质的编码基因可通过将SEQ ID NO:2所示的DNA序列中缺失一个或几个氨基酸残基的密码子，和/或进行一个或几个碱基对的错义突变，和/或在其5′端和/或3′端连上表1所示的标签的编码序列得到。
本发明还保护编码所述蛋白质Ms42的核酸分子。
所述编码所述蛋白质Ms42的核酸分子可为b1)或b2)或b3)或b4)所示的DNA分子：
b1)编码区是SEQ ID NO:2所示的DNA分子；
b2)核苷酸序列是SEQ ID NO:2所示的DNA分子；
b3)与b1)或b2)限定的核苷酸序列具有75％或75％以上同源性，来源于玉米且编码所述蛋白质Ms42的DNA分子；
b4)在严格条件下与b1)或b2)限定的核苷酸序列杂交，来源于玉米且编码所述蛋白质Ms42的DNA分子。
其中，所述核酸分子可以是DNA，如cDNA、基因组DNA或重组DNA；所述核酸分子也可以是RNA，如mRNA或hnRNA等。
其中，SEQ ID NO:2由2739个核苷酸组成，SEQ ID NO:2的核苷酸编码SEQ ID NO:1所示的氨基酸序列。
本领域普通技术人员可以很容易地采用已知的方法，例如定向进化和点突变的方法，对本发明的编码所述蛋白质Ms42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所述蛋白质Ms42的核苷酸序列75％或者更高同一性的核苷酸，只要编码所述蛋白质Ms42，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。
这里使用的术语“同一性”指与天然核酸序列的序列相似性。“同一性”包括与本发明的编码SEQ ID NO:1所示的氨基酸序列组成的蛋白质Ms42的核苷酸序列具有75％或更高，或80％或更高，或85％或更高，或90％或更高，或95％或更高同一性的核苷酸序列。同一性可以用肉眼或计算机软件进行评价。使用计算机软件，两个或多个序列之间的同一性可以用百分比(％)表示，其可以用来评价相关序列之间的同一性。
本发明还保护含有上述任一所述核酸分子的表达盒、重组载体或重组微生物。
所述含有上述任一所述核酸分子的重组载体可为在表达载体的多克隆位点插入SEQ ID NO：2所示的DNA分子得到的重组质粒。
所述含有上述任一所述核酸分子的重组微生物可为将含有上述任一所述核酸分子的重组载体导入出发微生物得到的重组菌。
所述出发微生物可为农杆菌或大肠杆菌。所述农杆菌具体可为根癌农杆菌。所述根癌农杆菌具体可为根癌农杆菌EHA105。
本发明还保护上述任一所述蛋白质Ms42、上述任一所述核酸分子或含有上述任一所述核酸分子的表达盒、重组载体或重组微生物，的应用，可为c1)或c2)：
c1)促进玉米雄性不育；
c2)培育雄性不育的玉米品种。
本发明还保护一种培育雄性不育玉米的方法，可包括如下步骤：提高雄性可育玉米中上述任一所述蛋白质Ms42的活性和/或表达量，获得转基因玉米甲；所述转基因玉米甲即为雄性不育玉米。
本发明还保护一种培育雄性可育玉米的方法，可包括如下步骤：抑制雄性不育玉米中上述任一所述蛋白质Ms42的活性和/或表达量，获得转基因玉米乙；所述转基因玉米乙即为雄性可育玉米；
所述雄性不育玉米的不育性状是由于表达上述任一所述蛋白质Ms42引起的。
上述方法中，所述“抑制雄性不育玉米中上述任一所述蛋白质Ms42的活性和/或表达量”具体可通过向雄性不育玉米中导入重组质粒实现；所述重组质粒的制备方法如下：将5’-GGCGTCCGCGCAGAAAAAACGCG-3’和5’-AAACCGCGTTTTTTCTGCGCGGA-3’进行退火反应，形成退火靶点片段，之后将pBUE411-BG载体和退火靶点片段连接，得到重组质粒。
上述方法中，所述雄性不育玉米可为玉米突变体Ms42。
所述转基因玉米乙可为实施例提及的Ms42-In1、Ms42-In2或Ms42-De2。Ms42-In1和Ms42-In2在Ms42基因和ms42基因的编码区分别发生了“A”1个核苷酸的插入和“TT”2个核苷酸的插入，从而引起了移码。Ms42-De2在Ms42基因和ms42基因的编码区发生了“TT”2个核苷酸的缺失，从而引起了移码。
实验证明，在玉米突变体Ms42中通过导入上述重组质粒抑制蛋白质Ms42的活性和/或表达量可以将玉米突变体Ms42的雄性不育恢复为雄性可育。Cas9培育雄性可育玉米突变株实验结果证实Ms42基因两个位点突变导致了玉米的显性雄性不育表型。因此，蛋白质Ms42及其编码基因可以促进玉米雄性不育，具有重要的应用价值。
本发明还保护蛋白质ms42，可为如下A1)或A2)或A3)或A4)：
A1)氨基酸序列是SEQ ID NO:3所示的蛋白质；
A2)在SEQ ID NO:3所示的蛋白质的N端或/和C端连接标签得到的融合蛋白质；
A3)将A1)或A2)所示的蛋白质经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的与玉米花粉耐热性相关的蛋白质；
A4)与SEQ ID NO:3限定的氨基酸序列具有80％或80％以上同源性，来源于玉米且与玉米花粉耐热性相关的蛋白质。
其中，SEQ ID NO:3由912个氨基酸残基组成。
为了使A1)中的蛋白质便于纯化，可在SEQ ID NO:3所示的蛋白质的氨基末端或羧基末端连接上如表1所示的标签。
上述A3)中的蛋白质，所述一个或几个氨基酸残基的取代和/或缺失和/或添加为不超过10个氨基酸残基的取代和/或缺失和/或添加。
上述A3)中的蛋白质可人工合成，也可先合成其编码基因，再进行生物表达得到。
上述A3)中的蛋白质的编码基因可通过将SEQ ID NO:4所示的DNA序列中缺失一个或几个氨基酸残基的密码子，和/或进行一个或几个碱基对的错义突变，和/或在其5′端和/或3′端连上表1所示的标签的编码序列得到。
本发明还保护编码所述蛋白质ms42的核酸分子。
所述编码所述蛋白质ms42的核酸分子可为B1)或B2)或B3)或B4)所示的DNA分子：
B1)编码区是SEQ ID NO:4所示的DNA分子；
B2)核苷酸序列是SEQ ID NO:4所示的DNA分子；
B3)与B1)或B2)限定的核苷酸序列具有75％或75％以上同源性，来源于玉米且编码所述蛋白质ms42的DNA分子；
B4)在严格条件下与B1)或B2)限定的核苷酸序列杂交，来源于玉米且编码所述蛋白质ms42的DNA分子。
其中，所述核酸分子可以是DNA，如cDNA、基因组DNA或重组DNA；所述核酸分子也可以是RNA，如mRNA或hnRNA等。
其中，SEQ ID NO:4由2739个核苷酸组成，SEQ ID NO:4的核苷酸编码SEQ ID NO:3所示的氨基酸序列。
本领域普通技术人员可以很容易地采用已知的方法，例如定向进化和点突变的方法，对本发明的编码所述蛋白质ms42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所述蛋白质ms42的核苷酸序列75％或者更高同一性的核苷酸，只要编码所述蛋白质ms42，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。
这里使用的术语“同一性”指与天然核酸序列的序列相似性。“同一性”包括与本发明的编码SEQ ID NO:3所示的氨基酸序列组成的蛋白质ms42的核苷酸序列具有75％或更高，或80％或更高，或85％或更高，或90％或更高，或95％或更高同一性的核苷酸序列。同一性可以用肉眼或计算机软件进行评价。使用计算机软件，两个或多个序列之间的同一性可以用百分比(％)表示，其可以用来评价相关序列之间的同一性。
本发明还保护含有编码所述蛋白质ms42的核酸分子的表达盒、重组载体或重组微生物。
所述含有编码所述蛋白质ms42的核酸分子的重组载体可为在表达载体的多克隆位点插入SEQ ID NO：4所示的DNA分子得到的重组质粒。
所述重组载体具体可为重组质粒pBCXUN-ms42。重组质粒pBCXUN-ms42为将植物表达载体pBCXUN-UbiP的限制性内切酶AscI和KpnI之间的DNA小片段替换为SEQ ID NO:4所示的DNA分子，得到的重组质粒。
所述含有编码所述蛋白质ms42的核酸分子的重组微生物可为将含有编码所述蛋白质ms42的核酸分子的重组载体(如重组质粒pBCXUN-ms42)导入出发微生物得到的重组菌。
所述出发微生物可为农杆菌或大肠杆菌。所述农杆菌具体可为根癌农杆菌。所述根癌农杆菌具体可为根癌农杆菌EHA105。
本发明还保护上述任一所述蛋白质ms42、编码所述蛋白质ms42的核酸分子或含有编码所述蛋白质ms42的核酸分子的表达盒、重组载体或重组微生物，的应用，可为C1)-C3)中的至少一种：
C1)提高玉米花粉耐热性；
C2)维持玉米雄性可育；
C3)培育花粉耐热的玉米品种。
本发明还保护一种降低玉米花粉耐热性方法，包括如下步骤：抑制出发玉米中所述蛋白质ms42的活性和/或表达量，获得转基因玉米丙；与出发玉米相比，转基因玉米丙的花粉耐热性降低。
本发明还保护一种降低玉米花粉耐热性方法，包括如下步骤：抑制出发玉米中所述蛋白质ms42的活性和/或表达量和所述蛋白质Ms42的活性和/或表达量，获得转基因玉米丁；与出发玉米相比，转基因玉米丁的花粉耐热性降低。
上述方法中，所述出发玉米可为玉米突变体Ms42。
所述转基因玉米丙或转基因玉米丁可为实施例提及的Ms42-In1、Ms42-In2或Ms42-De2。Ms42-In1和Ms42-In2在Ms42基因和ms42基因的编码区分别发生了“A”1个核苷酸的插入和“TT”2个核苷酸的插入，从而引起了移码。Ms42-De2在Ms42基因和ms42基因的编码区发生了“TT”2个核苷酸的缺失，从而引起了移码。
本发明还保护一种提高玉米花粉耐热性方法，可包括如下步骤：提高出发玉米中所述蛋白质ms42的活性和/或表达量，获得转基因玉米戊；与出发玉米相比，转基因玉米戊的花粉耐热性提高。
上述方法中，所述转基因玉米戊可为实施例提及的ms42-OE1或ms42-OE2。
上述方法中，所述“提高出发玉米中所述蛋白质ms42的活性和/或表达量”具体可通过向出发玉米中导入所述编码所述蛋白质ms42的核酸分子实现。所述编码所述蛋白质ms42的核酸分子具体可通过向出发玉米导入含有编码所述蛋白质ms42的核酸分子的重组载体实现。所述重组载体可为重组质粒pBCXUN-ms42。
实验证明，在玉米自交系B73中过表达ms42基因可以提高玉米花粉耐热性。ms42基因在提高花粉耐热性中具有重要的价值。由于花粉不耐热则花粉育性消失，因此，ms42基因在维持玉米雄性可育中具有重要的价值。
附图说明
图1为玉米突变体Ms42的表型鉴定结果。
图2为Ms42基因克隆和基因结构。A为Ms42的基因定位；B为Ms42的基因结构；C为Ms42的蛋白一级结构。
图3为Ms42-In1、Ms42-In2和Ms42-De2的突变类型和蛋白质Ms42含量的检测。
图4为Ms42-In1、Ms42-In2、Ms42-De2和玉米突变体Ms42的散粉期表型和花药育性鉴定。
图5为低热胁迫、中度热胁迫和高热胁迫下ms42-In1的雄穗表型和花药育性鉴定。
图6为低热胁迫、中度热胁迫和高热胁迫下ms42-In1的花粉育性统计结果。
图7为ms42-OE1和ms42-OE2中ms42基因的相对表达量检测及高热胁迫下WT、ms42-OE1和ms42-OE2的花粉育性统计结果。
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。
以下实施例中的定量试验，均设置三次重复实验，结果取平均值。
玉米突变体Ms42由美国Maize Genetics Cooperation Stock Center保存，Co-opID为502E；该玉米突变体在该保藏中心的具体网页链接如下：https://www.maizegdb.org/ data_center/stock？id＝142466.
pBUE411-BG载体记载于如下文献中：Xing，H.L.，Dong，L.，Wang，Z.P.，Zhang，H.Y.，Han，C.Y.，Liu,B.，Wang,X.C.，and Chen，Q.J.(2014).A CRISPR/Cas9toolkit formultiplex genome editing in plants.Bmc Plant Biology 14.pBUE411-BG载体具有抗膦丝菌素抗性(又称草胺磷抗性)。
玉米自交系B73-329记载于如下文献中：Zhang M，Cao Y，Wang Z，et al.Aretrotransposon in an HKT1 family sodium transporter causes variation of leafNa+exclusion and salt tolerance in maize[J].New Phytologist，2017.，在文献中的名称为玉米B73-329。
实施例1、与玉米雄性核不育相关的Ms42基因的图位克隆
一、玉米突变体Ms42的表型鉴定和花药育性鉴定
1、观察玉米突变体Ms42和玉米自交系B73(以下简称野生型或WT)的表型。
结果表明(见图1)，与野生型相比，玉米突变体Ms42表现出完全的雄性不育表型，但二者的营养生长状态无显著差异；散粉期，玉米突变体Ms42花药不能外露，花药更小；通过显微解剖，发现玉米突变体Ms42花药出现严重的萎缩。
2、花药育性鉴定
取玉米突变体Ms42或野生型的花药，碾碎(目的为获得花粉粒)，用1％I2-KI溶液染色。
结果表明，野生型的花粉粒为黑色且饱满，玉米突变体Ms42的花粉粒未被染成黑色且不规则(不是饱满状态)(见图1)。玉米突变体Ms42表现完全的雄性不育。
3、本发明的发明人用野生型的花粉对玉米突变体Ms42或野生型的雌穗进行授粉，观察结实情况。
结果表明，二者可以正常结实。
由此可见，玉米突变体Ms42的雌性育性正常，即雌配子发育正常。
二、玉米突变体Ms42遗传分析及与玉米雄性核不育相关的Ms42基因的图位克隆
1、为进一步研究玉米突变体Ms42的特性并获得目的基因，本发明的发明人以玉米突变体Ms42作为母本，以玉米自交系B73为父本进行杂交，得到杂交F1种子。
2、种植杂交F1种子，得到杂交F1植株。观90察杂交F1植株是否雄性可育。
结果表明，杂交F1植株表现出1:1育性分离(雄性可育：雄性不育＝134:142，χ2<χ2(0.05,1)＝3.84)。符合单基因显性突变的遗传定律。
由此可见，玉米突变体Ms42的雄性不育表型由一个显性基因控制。
3、本发明的发明人使用图位克隆方法来分离导致不育表型的基因。具体如下：
(1)将玉米突变体Ms42分别与玉米自交系B73和Zheng58杂交，产生了两个大的F1分离群体。连锁分析将位点定位在6号染色体的长臂上，通过使用两个背景分离群体的5000多株F1植物，将其进一步限定在两个InDel标记M2和M6之间的115kb区域。
(2)根据MaizeGDB(https://www.maizegdb.org/)的B73 v4参考基因组注释，在此区域内包含6个候选基因。(见图2中A)。
(3)为了进一步验证突变的基因，本发明的发明人分别使用雄性不育的亲本植物(即玉米突变体Ms42)和雄性可育的亲本植物(即玉米自交系B73)的减数分裂雄蕊提取RNA通过RNA-Seq分析这些基因的编码序列。
结果表明，雄性可育亲本和雄性不育亲本中存在一个候选基因。雄性不育亲本中该候选基因命名为Ms42基因，Ms42基因的核苷酸序列如SEQ ID NO:2所示，编码氨基酸序列如SEQ ID NO:1所示的蛋白质Ms42。雄性可育亲本中，该候选基因命名为ms42基因，ms42基因的核苷酸序列如SEQ ID NO:4所示，编码氨基酸序列如SEQ ID NO:3所示的蛋白质ms42。Ms42基因和ms42基因的不同之处在于：ms42基因第一个外显子的1072和1094位核苷酸处为鸟嘌呤(G)，而Ms42基因第一个外显子的1072和1094位核苷酸处为腺嘌呤(A)；两个单碱基突变分别导致了从谷氨酸(Glu)到赖氨酸(Lys)和从甘氨酸(Gly)到天冬氨酸(Asp)的两个氨基酸替换(图2中B和C)。发明人推测是ms42基因的这两个位点突变导致了玉米突变体Ms42玉米显性雄性不育的表型。
实施例2、培育雄性可育玉米突变株及其验证
选择一个靶点进行实验。靶点序列为：5’-GTCCGCGCAGAAAAAACGCGG-3’(即SEQ IDNo：2或SEQ ID No：4自5’末端起第297-316位)，对应靶基因为Ms42基因或ms42基因。
一、重组质粒的获得
1、人工合成引物42CAS9-F：5’-GGCGTCCGCGCAGAAAAAACGCG-3’和引物42CAS9-R：5’-AAACCGCGTTTTTTCTGCGCGGA-3’，用去离子水分别将引物42CAS9-F和引物42CAS9-R稀释至100μM，得到引物42CAS9-F稀释液和引物42CAS9-R稀释液；然后进行退火反应，形成退火靶点片段。
退火程序为：95℃10min，89℃7-8min，83℃7-8min，77℃7-8min，71℃7-8min，16℃保存。
2、将pBUE411-BG载体和退火靶点片段连接，得到重组质粒。
连接体系为15μL，由2μL退火靶点片段、2μL pBUE411-BG载体、1.5μL 10×NEB T4Buffer、1.5μL 10×BSA、1μL限制性内切酶BsaI、1μL T4 Ligase和6μL ddH2O组成。
连接程序为：37℃5h；50℃5min；80℃10min。
二、突变株的获得
1、将重组质粒通过液氮冷冻法转至根癌农杆菌EHA105，得到重组农杆菌。
2、将重组农杆菌转化至玉米突变体Ms42，经过筛选、分化和生根后，获得T0代拟转基因玉米。具体转化方法参考如下文献：魏晓禹，邵诗迪，孙苏等，农杆菌介导的玉米幼胚遗传转化体系的建立，吉林农业大学学报，2017(06)640-647；黄璐，卫志明，农杆菌介导的玉米遗传转化[J]，实验生物学报，1999(04).
3、分别以T0代拟转基因玉米叶片的基因组DNA为模板，采用引物F：5’-TAGCCGAAGCAAACCCATTCCAC-3’和引物R：5’-TTGTTCAGCACGCGCTCGAA-3’组成的引物对进行PCR扩增，得到相应的PCR扩增产物。
4、分别将PCR扩增产物进行Sanger测序。测序结果与SEQ ID No：2所示的Ms42基因Cas9目标序列进行比对，统计突变类型。
获得3个纯合突变株系(即在玉米两条染色体的同一位置发生相同的突变，本质就是Ms42基因和ms42基因均发生了移码，不再编码蛋白质Ms42和蛋白质ms42)，分别命名为Ms42-In1(即In1)、Ms42-In2(即In2)和Ms42-De2(即De2)。Ms42-In1和Ms42-In2在Ms42基因和ms42基因的编码区分别发生了“A”1个核苷酸的插入和“TT”2个核苷酸的插入，从而引起了移码；Ms42-De2在Ms42基因和ms42基因的编码区发生了“TT”2个核苷酸的缺失，从而引起了移码。突变类型具体见图3中A。
5、采用Western blot方法检测玉米自交系B73(简称WT)、Ms42-In1、Ms42-In2和Ms42-De2中蛋白质Ms42或蛋白质ms42的表达量，其中一抗为ms42特异性抗体(将ms42蛋白按照ABclonal公司流程进行免疫纯化抗体的制备)和Actin抗体(EASYBIO公司的产品，产品目录号为BE7008)，ms42特异性抗体用于检测蛋白质Ms42和蛋白质ms42，Actin抗体用于检测内参蛋白ZmActin蛋白。具体方法为：分别取玉米自交系B73、Ms42-In1、Ms42-In2和Ms42-De2的未成熟雄穗，液氮研磨提取总蛋白，之后采用ms42特异性抗体和Actin抗体westernblot检测。
检测结果见图3中B。结果表明，玉米自交系B73植株中可以检测到Ms42蛋白或ms42蛋白的条带，而Ms42-In1、Ms42-In2和Ms42-De2中无法检测到Ms42蛋白或ms42蛋白的条带。因此，3个纯合突变株系Ms42-In1、Ms42-In2和Ms42-De2均不编码蛋白质Ms42和蛋白质ms42。
三、突变株的表型鉴定和花药育性鉴定
1、观察玉米突变体Ms42、Ms42-In1、Ms42-In2和Ms42-De2散粉期的表型。
观察结果见图4中第一行。结果表明，散粉期，玉米突变体Ms42表现出完全的雄性不育表型、花药不能外露，Ms42-In1、Ms42-In2和Ms42-De2雄性可育，花药发育正常。玉米突变体Ms42、Ms42-In1、Ms42-In2和Ms42-De2的营养生长状态均无显著差异。
2、取玉米突变体Ms42、Ms42-In1、Ms42-In2或Ms42-De2的花药，碾碎(目的为获得花粉粒)，用1％I2-KI溶液染色。
结果见图4中第二行。结果表明，玉米突变体Ms42的花粉粒未被染成黑色且不规则(不是饱满状态)，Ms42-In1、Ms42-In2和Ms42-De2的花粉粒为黑色且饱满。
由此可见，玉米突变体Ms42表现完全的雄性不育，Ms42-In1、Ms42-In2和Ms42-De2恢复为雄性可育。
Cas9培育雄性可育玉米突变株实验结果证实Ms42基因两个位点突变导致了玉米的显性雄性不育表型。
实施例3、蛋白质ms42对花粉耐热性的影响
实验一、
分别将散粉期的玉米自交系B73(以下简称WT)和实施例2获得的Ms42-In1植株置于低热胁迫LHS(32℃处理14小时)、中度热胁迫MHS(35℃处理14小时)或高热胁迫HHS(35℃处理24小时)，观察表型并进行花药育性鉴定。进行花药育性鉴定的方法为：取花药，碾碎(目的为获得花粉粒)，用1％I2-KI溶液染色；被染成黑色的花粉粒可育，未被染成黑色的花粉粒不育；最后统计花粉育性，花粉育性＝被染成黑色的花粉粒数/(被染成黑色的花粉粒数+未被染成黑色的花粉粒数)×100％
玉米自交系B73和Ms42-In1均雄性可育。但玉米自交系B73可以表达蛋白质ms42，Ms42-In1不能表达蛋白质ms42，蛋白检测结果见图3中B。
高热胁迫处理后的表型见图5中右侧。结果表明，高热胁迫处理后，与WT相比，Ms42-In1花药不能外露；即高热胁迫处理后，Ms42-In1表现完全的雄性不育(Ms42-In1的花粉不耐热)。
花药育性鉴定结果见图5中左侧和图6。结果表明，LHS处理下，Ms42-In1与WT的花粉育性无显著差异；但在MHS和HHS处理下，与WT相比，Ms42-In1的花粉活力均显著下降(即Ms42-In1的花粉不耐热)。由此可见，Ms42-In1的花粉发育对高温胁迫比WT更敏感。
实验二、
(1)将植物表达载体pBCXUN-UbiP的限制性内切酶AscI和KpnI之间的DNA小片段替换为SEQ ID NO:4所示的DNA分子(即ms42基因)，得到重组质粒pBCXUN-ms42。
植物表达载体pBCXUN-UbiP为将pBCXUN载体(记载于如下文献中：Chen Songbiao，Songkumarn Pattavipha，Liu Jianli，et al.A versatile zero background T-vectorsystem for gene cloning and functional genomics.[J].Plant Physiol.，2009，150:1111-21.)中的HYG基因替换为Bar基因得到的重组质粒。植物表达载体pBCXUN-UbiP中，启动子为Ubiquitin启动子，终止子为T-nos终止子。植物表达载体pBCXUN-UbiP具有抗膦丝菌素抗性(又称草胺磷抗性)。
(2)将重组质粒pBCXUN-ms42通过液氮冷冻法转至根癌农杆菌EHA105，得到重组农杆菌。
(3)将重组农杆菌转化至玉米自交系B73(简称WT)，经过筛选、分化和生根后，获得拟转ms42基因玉米。具体转化方法参考如下文献：魏晓禹，邵诗迪，孙苏等，农杆菌介导的玉米幼胚遗传转化体系的建立，吉林农业大学学报，2017(06)640-647；黄璐，卫志明，农杆菌介导的玉米遗传转化[J]，实验生物学报，1999(04).
(4)分别提取各个拟转ms42基因玉米叶片的基因组DNA并作为模板，采用5’-CCTGAGCTGCTGAACCGTCT-3’和5’-TCGTCCATGCCGTCCTCGT-3’组成的引物对进行PCR扩增，得到PCR扩增产物。
(5)完成步骤(4)后，将各个PCR扩增产物进行1％(w/v)琼脂糖凝胶电泳，然后进行如下判断：如果以某拟转ms42基因玉米的基因组DNA为模板得到的PCR扩增产物中含有约473bp的DNA片段，则该拟转ms42基因玉米为转ms42基因玉米(阳性植株)；否则不为转ms42基因玉米。
结果表明，共获得转ms42基因玉米为10株，依次命名为ms42-OE1和ms42-OE10。
(6)提取玉米(ms42-OE1、ms42-OE2或玉米自交系B73)未成熟雄穗的总RNA，之后反转录为cDNA，实时定量PCR检测cDNA中ms42基因的相对表达量(以ZmActin(Zm00001d010159)基因为内参基因)。
检测ms42基因的引物为F：5’-TTCGATCCTCTGTCCCACGAG-3’和R：5’-AGACCGGCAACAGGATTCAATC-3’。
检测ZmActin基因的引物为5’-TCACCCTGTGCTGCTGACCG-3’和5’-GAACCGTGTGGCTCACACCA-3’。
实验重复三次取平均值。检测结果见图7中A。结果表明，与玉米自交系B73相比，ms42-OE1和ms42-OE2中ms42基因的相对表达量均显著增加。
分别将散粉期的玉米自交系B73(以下简称WT)、ms42-OE1和ms42-OE2植株置于常规处理NT(28℃处理16小时或24小时)或更高热胁迫HHT(38℃处理16小时或24小时)，观察表型并进行花药育性鉴定。进行花药育性鉴定的方法为：取花药，碾碎(目的为获得花粉粒)，用1％I2-KI溶液染色；被染成黑色的花粉粒可育，未被染成黑色的花粉粒不育；最后统计花粉育性，花粉育性＝被染成黑色的花粉粒数/(被染成黑色的花粉粒数+未被染成黑色的花粉粒数)×100％
花药育性鉴定结果见图7中B。花粉育性统计结果见图7中C。结果表明，常规处理时，WT、ms42-OE1和ms42-OE2的花粉育性无显著差异；38℃高热胁迫处理16小时或24小时后，与WT相比，ms42-OE1和ms42-OE2的花粉育性均显著提高。由此可见，与WT相比，ms42-OE1和ms42-OE2的花粉更具有耐热性。
上述结果表明，ms42基因在提高花粉耐热性中具有重要的价值。由于花粉不耐热则花粉育性消失，因此，ms42基因在维持玉米雄性可育中具有重要的价值。
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。
<110>中国农业大学
<120>调控玉米花粉育性和花粉耐热性的方法
<160>4
<170>PatentIn version 3.5
<210>1
<211>912
<212>PRT
<213>Artificial sequence
<400>1
Met Asn Pro Asp Asn Phe Thr His Lys Thr Asn Glu Ala Ile Val Gly
1 5 10 15
Ala His Glu Ile Ala Val Glu Ala Gly His Ala Gln Leu Thr Pro Leu
20 25 30
His Leu Ala Ala Val Leu Ala Ala Asp Lys Gly Gly Ile Leu Arg Gln
35 40 45
Ala Ile Thr Gly Ala Ser Gly Gly Asp Gly Ala Ala Gly Asp Ser Phe
50 55 60
Glu Arg Val Leu Asn Asn Ser Leu Lys Lys Leu Pro Ser Gln Ser Pro
65 70 75 80
Pro Pro Asp Ser Val Pro Ala Ser Thr Ala Leu Ile Lys Val Ile Arg
85 90 95
Arg Ala Gln Ser Ala Gln Lys Lys Arg Gly Asp Ser His Leu Ala Val
100 105 110
Asp Gln Leu Leu Leu Gly Leu Leu Glu Asp Ser Gln Ile Ser Asp Cys
115 120 125
Leu Lys Glu Ala Gly Val Ser Ala Ala Arg Val Arg Ala Glu Leu Glu
130 135 140
Lys Leu Arg Gly Gly Glu Gly Arg Arg Val Glu Ser Ala Ser Gly Asp
145 150 155 160
Thr Asn Phe Gln Ala Leu Lys Thr Tyr Gly Arg Asp Leu Val Glu Gln
165 170 175
Ala Gly Lys Leu Asp Pro Val Ile Gly Arg Asp Glu Glu Ile Arg Arg
180 185 190
Val Val Arg Ile Leu Ser Arg Arg Thr Lys Asn Asn Pro Val Leu Ile
195 200 205
Gly Glu Pro Gly Val Gly Lys Thr Ala Val Val Glu Gly Leu Ala Gln
210 215 220
Arg Ile Val Arg Gly Asp Val Pro Ser Asn Leu Leu Asp Val Arg Leu
225 230 235 240
Ile Ala Leu Asp Met Gly Ala Leu Val Ala Gly Ala Lys Tyr Arg Gly
245 250 255
Glu Phe Glu Glu Arg Leu Lys Ala Val Leu Lys Glu Val Glu Glu Ala
260 265 270
Glu Gly Lys Val Ile Leu Phe Ile Asp Glu Ile His Leu Val Leu Gly
275 280 285
Ala Gly Arg Thr Glu Gly Ser Met Asp Ala Ala Asn Leu Phe Lys Pro
290 295 300
Met Leu Ala Arg Gly Gln Leu Arg Cys Ile Gly Ala Thr Thr Leu Glu
305 310 315 320
Glu Tyr Arg Lys Tyr Val Glu Lys Asp Ala Ala Phe Glu Arg Arg Phe
325 330 335
Gln Gln Val Phe Val Ala Glu Pro Ser Val Pro Asp Thr Val Ser Ile
340 345 350
Leu Arg Gly Leu Lys Lys Lys Tyr Glu Gly His His Asp Val Arg Ile
355 360 365
Gln Asp Arg Ala Leu Val Val Ala Ala Gln Leu Ser Ala Arg Tyr Ile
370 375 380
Met Gly Arg His Leu Pro Asp Lys Ala Ile Asp Leu Val Asp Glu Ala
385 390 395 400
Cys Ala Asn Val Arg Val Gln Leu Asp Ser Gln Pro Glu Glu Ile Asp
405 410 415
Asn Leu Glu Arg Lys Arg Ile Gln Leu Glu Val Glu Leu His Ala Leu
420 425 430
Glu Lys Glu Lys Asp Lys Ala Ser Lys Ala Arg Leu Ile Glu Val Arg
435 440 445
Lys Glu Leu Asp Asp Leu Arg Asp Lys Leu Gln Pro Leu Thr Met Lys
450 455 460
Tyr Arg Lys Glu Lys Glu Arg Ile Asp Glu Ile Arg Lys Leu Lys Gln
465 470 475 480
Arg Arg Glu Glu Leu Gln Phe Thr Leu Gln Glu Ala Glu Arg Arg Met
485 490 495
Asp Leu Ala Arg Val Ala Asp Leu Lys Tyr Gly Ala Leu Gln Glu Ile
500 505 510
Asp Ala Ala Ile Ser Lys Leu Glu Ser Glu Thr Gly Glu Asn Leu Met
515 520 525
Leu Thr Glu Thr Val Gly Pro Glu Gln Ile Ala Glu Val Val Ser Arg
530 535 540
Trp Thr Gly Ile Pro Val Thr Arg Leu Gly Gln Asn Asp Lys Glu Arg
545 550 555 560
Leu Val Gly Leu Ala Asp Arg Leu His Gln Arg Val Val Gly Gln Thr
565 570 575
Glu Ala Val Ser Ala Val Ala Glu Ala Val Leu Arg Ser Arg Ala Gly
580 585 590
Leu Gly Arg Pro Gln Gln Pro Thr Gly Ser Phe Leu Phe Leu Gly Pro
595 600 605
Thr Gly Val Gly Lys Thr Glu Leu Ala Lys Ala Leu Ala Glu Gln Leu
610 615 620
Phe Asp Asp Glu Asn Leu Leu Val Arg Ile Asp Met Ser Glu Tyr Met
625 630 635 640
Glu Gln His Ser Val Ala Arg Leu Ile Gly Ala Pro Pro Gly Tyr Val
645 650 655
Gly His Glu Glu Gly Gly Gln Leu Thr Glu Gln Val Arg Arg Arg Pro
660 665 670
Tyr Ser Val Ile Leu Phe Asp Glu Val Glu Lys Ala His Val Ala Val
675 680 685
Phe Asn Thr Leu Leu Gln Val Leu Asp Asp Gly Arg Leu Thr Asp Gly
690 695 700
Gln Gly Arg Thr Val Asp Phe Arg Asn Thr Val Ile Ile Met Thr Ser
705 710 715 720
Asn Leu Gly Ala Glu His Leu Leu Ala Gly Met Val Gly Lys Asn Ser
725 730 735
Met Lys Val Ala Arg Asp Leu Val Met Gln Glu Val Arg Arg His Phe
740 745 750
Arg Pro Glu Leu Leu Asn Arg Leu Asp Glu Ile Val Ile Phe Asp Pro
755 760 765
Leu Ser His Glu Gln Leu Arg Lys Val Ala Arg Leu Gln Met Lys Asp
770 775 780
Val Ala Val Arg Leu Ala Glu Arg Gly Ile Ala Leu Ala Val Thr Asp
785 790 795 800
Ala Ala Leu Asp Ile Ile Leu Ser Leu Ser Tyr Asp Pro Val Tyr Gly
805 810 815
Ala Arg Pro Ile Arg Arg Trp Ile Glu Lys Arg Val Val Thr Gln Leu
820 825 830
Ser Lys Met Leu Ile Gln Glu Glu Ile Asp Glu Asn Cys Thr Val Tyr
835 840 845
Ile Asp Ala Ala Pro Gly Lys Asp Glu Leu Val Tyr Arg Val Asp Arg
850 855 860
Ser Gly Gly Leu Val Asn Ala Glu Thr Gly Met Lys Ser Asp Ile Leu
865 870 875 880
Ile Gln Val Pro Thr Ser Ser Thr Arg Ser Asp Ala Ala Gln Ala Val
885 890 895
Lys Lys Met Arg Ile Met Glu Glu Asp Glu Asp Gly Met Asp Glu Glu
900 905 910
<210>2
<211>2739
<212>DNA
<213>Artificial sequence
<400>2
atgaatccgg acaacttcac ccacaagacg aacgaggcga tcgtgggggc gcacgagatt 60
gcggtggagg ccggccacgc gcagctcacg ccgctgcacc tggccgcagt gctggctgcg 120
gacaagggcg gcatcctgcg gcaggccatc acgggggcgt cggggggcga cggagcggcc 180
ggggactcgt tcgagcgcgt gctgaacaac tcgctcaaga agctgccgtc gcagtccccg 240
ccgccggact ccgtcccggc gtccacggca ctgatcaagg tcatccgccg ggcgcagtcc 300
gcgcagaaaa aacgcgggga ctctcacctc gccgtcgacc agctgctgct cggcctgctc 360
gaggactcgc agatctccga ctgcctcaag gaggccggcg tgtccgcggc gcgggtgcgc 420
gccgagcttg agaagctccg cggcggggag gggcgccgcg tggagtccgc gtcgggggac 480
actaacttcc aggcgctcaa gacatacggc cgggacctcg tcgagcaggc cgggaagctt 540
gatcccgtca tcggccgcga cgaggagatc cgccgcgtgg tgcgcatcct ctcgcgccgc 600
actaagaaca accccgtcct catcggcgag cccggcgttg gcaagacggc cgtcgtggag 660
ggcctcgcgc agcgcatcgt tcgcggcgac gtgcccagta acctcctcga cgtccgcctc 720
atcgcgctcg acatgggcgc gcttgtcgcg ggcgccaagt accgcggcga gttcgaggag 780
cggctcaagg ccgtgctcaa ggaggtggaa gaggccgagg ggaaggtcat tctcttcatc 840
gacgagatac acctcgtcct gggcgcgggc aggacggagg gttccatgga cgcggccaac 900
ctgttcaagc caatgctggc gaggggacag ctcaggtgca tcggcgccac cacgctggag 960
gagtaccgca agtacgtgga gaaggacgca gcgttcgagc ggcggttcca gcaggtgttc 1020
gtcgcggagc cgagcgtgcc cgataccgtc agcattctga ggggactcaa gaagaagtac 1080
gaggggcacc atgacgtgag gatccaggac cgcgccctcg tggtcgcggc acagctctcc 1140
gcgaggtaca tcatgggtcg gcacctgcct gacaaagcca tagacctggt ggacgaggcc 1200
tgcgccaatg tgagggtgca gctcgacagc cagccggagg agattgataa cctggagagg 1260
aagagaatcc agcttgaggt cgagctccac gcgctcgaga aggagaagga caaggccagt 1320
aaagcccggc tgattgaggt caggaaggaa ttggacgatc tgagggacaa gctgcagccc 1380
ctgaccatga agtacaggaa ggagaaggag agaatcgatg agatcaggaa gctgaagcag 1440
cgccgcgagg agctccagtt caccctgcag gaggccgagc gccggatgga cctggcccgt 1500
gtggccgatc tcaagtacgg cgccctccag gaaatcgacg ccgcgatctc caagctggag 1560
agcgaaacag gggagaacct gatgctcacc gaaaccgtcg gccctgaaca aattgcagag 1620
gtggtgagcc gttggacggg tattccagtg acccggcttg gccagaacga caaggagagg 1680
ctggtcggcc tggctgacag gcttcaccag agggtggtcg gccagacaga ggctgtgagc 1740
gccgtcgcag aggcggtgct gaggtcgagg gccggtcttg gcaggccaca acagcccact 1800
ggctcgttcc tcttcctggg tccgactggc gtggggaaaa ctgagctggc caaggcccta 1860
gccgaacagc tgttcgacga cgagaacctt cttgtccgca tcgacatgtc ggagtacatg 1920
gagcagcact cggttgctcg cctcatcgga gcaccacctg gctacgtcgg ccatgaagag 1980
ggtgggcagc tgactgaaca agtgaggagg aggccgtaca gcgtgatcct gttcgacgag 2040
gtcgagaagg cccatgtcgc cgtgttcaac accctgctcc aggtcctcga cgacggcagg 2100
ctgacggatg ggcaaggcag gacggtggac ttcaggaaca ccgtgatcat catgacatcg 2160
aacctcggcg ccgagcacct cctcgctggg atggtgggca agaactccat gaaggtcgct 2220
cgcgatctgg tcatgcagga ggtgaggagg cacttccgcc ctgagctgct gaaccgtctc 2280
gacgagatcg tgatcttcga tcctctgtcc cacgagcagc tgaggaaggt cgctcgcctt 2340
cagatgaagg atgtggccgt ccgtcttgcc gaaaggggca tcgctctggc tgtgaccgac 2400
gccgcattgg acatcatctt gtctctctct tacgatccgg tgtacggcgc gcggccaata 2460
aggaggtgga tcgagaagag ggtggtgacg cagctgtcga agatgctgat ccaggaggag 2520
atcgacgaga actgcacggt ctacatcgac gccgcgcccg gtaaggacga actggtctac 2580
agggtggacc ggagcggcgg tctggtgaac gctgagacgg ggatgaagtc ggacatcctg 2640
atccaggtcc ccaccagctc caccaggagc gacgctgcgc aggccgtcaa gaagatgagg 2700
atcatggagg aggacgagga cggcatggac gaggagtaa 2739
<210>3
<211>912
<212>PRT
<213>Artificial sequence
<400>3
Met Asn Pro Asp Asn Phe Thr His Lys Thr Asn Glu Ala Ile Val Gly
1 5 10 15
Ala His Glu Ile Ala Val Glu Ala Gly His Ala Gln Leu Thr Pro Leu
20 25 30
His Leu Ala Ala Val Leu Ala Ala Asp Lys Gly Gly Ile Leu Arg Gln
35 40 45
Ala Ile Thr Gly Ala Ser Gly Gly Asp Gly Ala Ala Gly Asp Ser Phe
50 55 60
Glu Arg Val Leu Asn Asn Ser Leu Lys Lys Leu Pro Ser Gln Ser Pro
65 70 75 80
Pro Pro Asp Ser Val Pro Ala Ser Thr Ala Leu Ile Lys Val Ile Arg
85 90 95
Arg Ala Gln Ser Ala Gln Lys Lys Arg Gly Asp Ser His Leu Ala Val
100 105 110
Asp Gln Leu Leu Leu Gly Leu Leu Glu Asp Ser Gln Ile Ser Asp Cys
115 120 125
Leu Lys Glu Ala Gly Val Ser Ala Ala Arg Val Arg Ala Glu Leu Glu
130 135 140
Lys Leu Arg Gly Gly Glu Gly Arg Arg Val Glu Ser Ala Ser Gly Asp
145 150 155 160
Thr Asn Phe Gln Ala Leu Lys Thr Tyr Gly Arg Asp Leu Val Glu Gln
165 170 175
Ala Gly Lys Leu Asp Pro Val Ile Gly Arg Asp Glu Glu Ile Arg Arg
180 185 190
Val Val Arg Ile Leu Ser Arg Arg Thr Lys Asn Asn Pro Val Leu Ile
195 200 205
Gly Glu Pro Gly Val Gly Lys Thr Ala Val Val Glu Gly Leu Ala Gln
210 215 220
Arg Ile Val Arg Gly Asp Val Pro Ser Asn Leu Leu Asp Val Arg Leu
225 230 235 240
Ile Ala Leu Asp Met Gly Ala Leu Val Ala Gly Ala Lys Tyr Arg Gly
245 250 255
Glu Phe Glu Glu Arg Leu Lys Ala Val Leu Lys Glu Val Glu Glu Ala
260 265 270
Glu Gly Lys Val Ile Leu Phe Ile Asp Glu Ile His Leu Val Leu Gly
275 280 285
Ala Gly Arg Thr Glu Gly Ser Met Asp Ala Ala Asn Leu Phe Lys Pro
290 295 300
Met Leu Ala Arg Gly Gln Leu Arg Cys Ile Gly Ala Thr Thr Leu Glu
305 310 315 320
Glu Tyr Arg Lys Tyr Val Glu Lys Asp Ala Ala Phe Glu Arg Arg Phe
325 330 335
Gln Gln Val Phe Val Ala Glu Pro Ser Val Pro Asp Thr Val Ser Ile
340 345 350
Leu Arg Gly Leu Lys Glu Lys Tyr Glu Gly His His Gly Val Arg Ile
355 360 365
Gln Asp Arg Ala Leu Val Val Ala Ala Gln Leu Ser Ala Arg Tyr Ile
370 375 380
Met Gly Arg His Leu Pro Asp Lys Ala Ile Asp Leu Val Asp Glu Ala
385 390 395 400
Cys Ala Asn Val Arg Val Gln Leu Asp Ser Gln Pro Glu Glu Ile Asp
405 410 415
Asn Leu Glu Arg Lys Arg Ile Gln Leu Glu Val Glu Leu His Ala Leu
420 425 430
Glu Lys Glu Lys Asp Lys Ala Ser Lys Ala Arg Leu Ile Glu Val Arg
435 440 445
Lys Glu Leu Asp Asp Leu Arg Asp Lys Leu Gln Pro Leu Thr Met Lys
450 455 460
Tyr Arg Lys Glu Lys Glu Arg Ile Asp Glu Ile Arg Lys Leu Lys Gln
465 470 475 480
Arg Arg Glu Glu Leu Gln Phe Thr Leu Gln Glu Ala Glu Arg Arg Met
485 490 495
Asp Leu Ala Arg Val Ala Asp Leu Lys Tyr Gly Ala Leu Gln Glu Ile
500 505 510
Asp Ala Ala Ile Ser Lys Leu Glu Ser Glu Thr Gly Glu Asn Leu Met
515 520 525
Leu Thr Glu Thr Val Gly Pro Glu Gln Ile Ala Glu Val Val Ser Arg
530 535 540
Trp Thr Gly Ile Pro Val Thr Arg Leu Gly Gln Asn Asp Lys Glu Arg
545 550 555 560
Leu Val Gly Leu Ala Asp Arg Leu His Gln Arg Val Val Gly Gln Thr
565 570 575
Glu Ala Val Ser Ala Val Ala Glu Ala Val Leu Arg Ser Arg Ala Gly
580 585 590
Leu Gly Arg Pro Gln Gln Pro Thr Gly Ser Phe Leu Phe Leu Gly Pro
595 600 605
Thr Gly Val Gly Lys Thr Glu Leu Ala Lys Ala Leu Ala Glu Gln Leu
610 615 620
Phe Asp Asp Glu Asn Leu Leu Val Arg Ile Asp Met Ser Glu Tyr Met
625 630 635 640
Glu Gln His Ser Val Ala Arg Leu Ile Gly Ala Pro Pro Gly Tyr Val
645 650 655
Gly His Glu Glu Gly Gly Gln Leu Thr Glu Gln Val Arg Arg Arg Pro
660 665 670
Tyr Ser Val Ile Leu Phe Asp Glu Val Glu Lys Ala His Val Ala Val
675 680 685
Phe Asn Thr Leu Leu Gln Val Leu Asp Asp Gly Arg Leu Thr Asp Gly
690 695 700
Gln Gly Arg Thr Val Asp Phe Arg Asn Thr Val Ile Ile Met Thr Ser
705 710 715 720
Asn Leu Gly Ala Glu His Leu Leu Ala Gly Met Val Gly Lys Asn Ser
725 730 735
Met Lys Val Ala Arg Asp Leu Val Met Gln Glu Val Arg Arg His Phe
740 745 750
Arg Pro Glu Leu Leu Asn Arg Leu Asp Glu Ile Val Ile Phe Asp Pro
755 760 765
Leu Ser His Glu Gln Leu Arg Lys Val Ala Arg Leu Gln Met Lys Asp
770 775 780
Val Ala Val Arg Leu Ala Glu Arg Gly Ile Ala Leu Ala Val Thr Asp
785 790 795 800
Ala Ala Leu Asp Ile Ile Leu Ser Leu Ser Tyr Asp Pro Val Tyr Gly
805 810 815
Ala Arg Pro Ile Arg Arg Trp Ile Glu Lys Arg Val Val Thr Gln Leu
820 825 830
Ser Lys Met Leu Ile Gln Glu Glu Ile Asp Glu Asn Cys Thr Val Tyr
835 840 845
Ile Asp Ala Ala Pro Gly Lys Asp Glu Leu Val Tyr Arg Val Asp Arg
850 855 860
Ser Gly Gly Leu Val Asn Ala Glu Thr Gly Met Lys Ser Asp Ile Leu
865 870 875 880
Ile Gln Val Pro Thr Ser Ser Thr Arg Ser Asp Ala Ala Gln Ala Val
885 890 895
Lys Lys Met Arg Ile Met Glu Glu Asp Glu Asp Gly Met Asp Glu Glu
900 905 910
<210>4
<211>2739
<212>DNA
<213>Artificial sequence
<400>4
atgaatccgg acaacttcac ccacaagacg aacgaggcga tcgtgggggc gcacgagatt 60
gcggtggagg ccggccacgc gcagctcacg ccgctgcacc tggccgcagt gctggctgcg 120
gacaagggcg gcatcctgcg gcaggccatc acgggggcgt cggggggcga cggagcggcc 180
ggggactcgt tcgagcgcgt gctgaacaac tcgctcaaga agctgccgtc gcagtccccg 240
ccgccggact ccgtcccggc gtccacggca ctgatcaagg tcatccgccg ggcgcagtcc 300
gcgcagaaaa aacgcgggga ctctcacctc gccgtcgacc agctgctgct cggcctgctc 360
gaggactcgc agatctccga ctgcctcaag gaggccggcg tgtccgcggc gcgggtgcgc 420
gccgagcttg agaagctccg cggcggggag gggcgccgcg tggagtccgc gtcgggggac 480
actaacttcc aggcgctcaa gacatacggc cgggacctcg tcgagcaggc cgggaagctt 540
gatcccgtca tcggccgcga cgaggagatc cgccgcgtgg tgcgcatcct ctcgcgccgc 600
actaagaaca accccgtcct catcggcgag cccggcgttg gcaagacggc cgtcgtggag 660
ggcctcgcgc agcgcatcgt tcgcggcgac gtgcccagta acctcctcga cgtccgcctc 720
atcgcgctcg acatgggcgc gcttgtcgcg ggcgccaagt accgcggcga gttcgaggag 780
cggctcaagg ccgtgctcaa ggaggtggaa gaggccgagg ggaaggtcat tctcttcatc 840
gacgagatac acctcgtcct gggcgcgggc aggacggagg gttccatgga cgcggccaac 900
ctgttcaagc caatgctggc gaggggacag ctcaggtgca tcggcgccac cacgctggag 960
gagtaccgca agtacgtgga gaaggacgca gcgttcgagc ggcggttcca gcaggtgttc 1020
gtcgcggagc cgagcgtgcc cgataccgtc agcattctga ggggactcaa ggagaagtac 1080
gaggggcacc atggcgtgag gatccaggac cgcgccctcg tggtcgcggc acagctctcc 1140
gcgaggtaca tcatgggtcg gcacctgcct gacaaagcca tagacctggt ggacgaggcc 1200
tgcgccaatg tgagggtgca gctcgacagc cagccggagg agattgataa cctggagagg 1260
aagagaatcc agcttgaggt cgagctccac gcgctcgaga aggagaagga caaggccagt 1320
aaagcccggc tgattgaggt caggaaggaa ttggacgatc tgagggacaa gctgcagccc 1380
ctgaccatga agtacaggaa ggagaaggag agaatcgatg agatcaggaa gctgaagcag 1440
cgccgcgagg agctccagtt caccctgcag gaggccgagc gccggatgga cctggcccgt 1500
gtggccgatc tcaagtacgg cgccctccag gaaatcgacg ccgcgatctc caagctggag 1560
agcgaaacag gggagaacct gatgctcacc gaaaccgtcg gccctgaaca aattgcagag 1620
gtggtgagcc gttggacggg tattccagtg acccggcttg gccagaacga caaggagagg 1680
ctggtcggcc tggctgacag gcttcaccag agggtggtcg gccagacaga ggctgtgagc 1740
gccgtcgcag aggcggtgct gaggtcgagg gccggtcttg gcaggccaca acagcccact 1800
ggctcgttcc tcttcctggg tccgactggc gtggggaaaa ctgagctggc caaggcccta 1860
gccgaacagc tgttcgacga cgagaacctt cttgtccgca tcgacatgtc ggagtacatg 1920
gagcagcact cggttgctcg cctcatcgga gcaccacctg gctacgtcgg ccatgaagag 1980
ggtgggcagc tgactgaaca agtgaggagg aggccgtaca gcgtgatcct gttcgacgag 2040
gtcgagaagg cccatgtcgc cgtgttcaac accctgctcc aggtcctcga cgacggcagg 2100
ctgacggatg ggcaaggcag gacggtggac ttcaggaaca ccgtgatcat catgacatcg 2160
aacctcggcg ccgagcacct cctcgctggg atggtgggca agaactccat gaaggtcgct 2220
cgcgatctgg tcatgcagga ggtgaggagg cacttccgcc ctgagctgct gaaccgtctc 2280
gacgagatcg tgatcttcga tcctctgtcc cacgagcagc tgaggaaggt cgctcgcctt 2340
cagatgaagg atgtggccgt ccgtcttgcc gaaaggggca tcgctctggc tgtgaccgac 2400
gccgcattgg acatcatctt gtctctctct tacgatccgg tgtacggcgc gcggccaata 2460
aggaggtgga tcgagaagag ggtggtgacg cagctgtcga agatgctgat ccaggaggag 2520
atcgacgaga actgcacggt ctacatcgac gccgcgcccg gtaaggacga actggtctac 2580
agggtggacc ggagcggcgg tctggtgaac gctgagacgg ggatgaagtc ggacatcctg 2640
atccaggtcc ccaccagctc caccaggagc gacgctgcgc aggccgtcaa gaagatgagg 2700
atcatggagg aggacgagga cggcatggac gaggagtaa 2739
Claims (7)
1.蛋白质Ms42，为如下a1)或a2)：
a1)氨基酸序列是SEQ ID NO:1所示的蛋白质；
a2)在SEQ ID NO:1所示的蛋白质的N端或/和C端连接标签得到的融合蛋白质。
2.编码权利要求1所述蛋白质Ms42的核酸分子。
3.根据权利要求2所述的核酸分子，其特征在于：所述核酸分子为b1)或b2)所示的DNA分子：
b1)编码区是SEQ ID NO:2所示的DNA分子；
b2)核苷酸序列是SEQ ID NO:2所示的DNA分子。
4.含有权利要求2或3所述核酸分子的表达盒、重组载体或重组微生物。
5.权利要求1所述蛋白质Ms42、权利要求2或3所述核酸分子或含有权利要求2或3所述核酸分子的表达盒、重组载体或重组微生物的应用，为c1)或c2)：
c1)促进玉米雄性不育；
c2)培育雄性不育的玉米品种。
6.一种培育雄性不育玉米的方法，包括如下步骤：提高雄性可育玉米中权利要求1所述蛋白质Ms42的活性和/或表达量，获得转基因玉米甲；所述转基因玉米甲即为雄性不育玉米。
7.一种培育雄性可育玉米的方法，包括如下步骤：抑制雄性不育玉米中权利要求1所述蛋白质Ms42的活性和/或表达量，获得转基因玉米乙；所述转基因玉米乙即为雄性可育玉米；
所述雄性不育玉米的不育性状是由于表达权利要求1所述蛋白质Ms42引起的。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210180293.0A CN114853857B (zh) | 2022-02-25 | 2022-02-25 | 调控玉米花粉育性和花粉耐热性的方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210180293.0A CN114853857B (zh) | 2022-02-25 | 2022-02-25 | 调控玉米花粉育性和花粉耐热性的方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114853857A CN114853857A (zh) | 2022-08-05 |
| CN114853857B true CN114853857B (zh) | 2024-01-26 |
Family
ID=82628427
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210180293.0A Active CN114853857B (zh) | 2022-02-25 | 2022-02-25 | 调控玉米花粉育性和花粉耐热性的方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114853857B (zh) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104946660A (zh) * | 2014-11-30 | 2015-09-30 | 北京首佳利华科技有限公司 | 一种玉米花粉减数分裂后发育调控基因Ms30的DNA序列及其编码蛋白 |
| CN107176981A (zh) * | 2017-06-26 | 2017-09-19 | 中国科学院遗传与发育生物学研究所 | Meica1蛋白及其编码基因在调控花粉育性中的应用 |
| CN109207513A (zh) * | 2018-10-22 | 2019-01-15 | 中国科学院遗传与发育生物学研究所 | Dcm1蛋白及其编码基因在调控植物雄性育性中的应用 |
| CN109439667A (zh) * | 2017-09-30 | 2019-03-08 | 海南波莲水稻基因科技有限公司 | 玉米基因ZmABCG20在调控作物雄性育性中的应用 |
| CN111909250A (zh) * | 2019-05-10 | 2020-11-10 | 中国农业大学 | 蛋白质invan6、其编码基因以及它们在选育玉米雄性不育系中的应用 |
| CN113943356A (zh) * | 2021-01-26 | 2022-01-18 | 中国农业大学 | 蛋白质phd11、其编码基因以及它们在选育玉米雄性不育系中的应用 |
| CN113980996A (zh) * | 2021-11-16 | 2022-01-28 | 中国农业大学 | 蛋白gen1及其相关生物材料在调控玉米产量中的应用 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AR078696A1 (es) * | 2009-08-04 | 2011-11-30 | Evogene Ltd | Polinucleotidos y polipeptidos aislados, y metodos para utilizarlos para mejorar la tolerancia al estres abiotico, rendimiento, tasa de crecimiento, vigor, biomasa, contenido de aceite y/o eficacia en el uso de nitrogeno de las plantas |
-
2022
- 2022-02-25 CN CN202210180293.0A patent/CN114853857B/zh active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104946660A (zh) * | 2014-11-30 | 2015-09-30 | 北京首佳利华科技有限公司 | 一种玉米花粉减数分裂后发育调控基因Ms30的DNA序列及其编码蛋白 |
| CN107176981A (zh) * | 2017-06-26 | 2017-09-19 | 中国科学院遗传与发育生物学研究所 | Meica1蛋白及其编码基因在调控花粉育性中的应用 |
| CN109439667A (zh) * | 2017-09-30 | 2019-03-08 | 海南波莲水稻基因科技有限公司 | 玉米基因ZmABCG20在调控作物雄性育性中的应用 |
| CN109207513A (zh) * | 2018-10-22 | 2019-01-15 | 中国科学院遗传与发育生物学研究所 | Dcm1蛋白及其编码基因在调控植物雄性育性中的应用 |
| CN111909250A (zh) * | 2019-05-10 | 2020-11-10 | 中国农业大学 | 蛋白质invan6、其编码基因以及它们在选育玉米雄性不育系中的应用 |
| CN113943356A (zh) * | 2021-01-26 | 2022-01-18 | 中国农业大学 | 蛋白质phd11、其编码基因以及它们在选育玉米雄性不育系中的应用 |
| CN113980996A (zh) * | 2021-11-16 | 2022-01-28 | 中国农业大学 | 蛋白gen1及其相关生物材料在调控玉米产量中的应用 |
Non-Patent Citations (2)
| Title |
|---|
| NIETO-SOTELO, J等.Maize HSP101 Plays Important Roles in Both Induced and Basal Thermotolerance and Primary Root Growth.Plant Cell.2002,第127卷(第3期),摘要，第1630页左栏第二段，第1632页左栏第三段. * |
| YOUNG, T.E.等.Developmental and Thermal Regulation of the Maize Heat Shock Protein, HSP101.Plant Physiology.2001,第127卷(第3期),摘要. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114853857A (zh) | 2022-08-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3018217B1 (en) | Maize cytoplasmic male sterility (cms) c-type restorer rf4 gene, molecular markers and their use | |
| CN110205327B (zh) | 一种水稻温敏核不育基因tms3突变体及其分子标记与应用 | |
| WO2018019193A1 (zh) | 小麦育性恢复基因及其应用 | |
| CN101698677B (zh) | 一种植物株高相关蛋白及其编码基因与应用 | |
| CN101704881B (zh) | 一种植物雄性育性相关蛋白及其编码基因与应用 | |
| CN111153974A (zh) | 玉米抗病基因和分子标记及其应用 | |
| CN107759676B (zh) | 一种植物直链淀粉合成相关蛋白Du15与其编码基因及应用 | |
| US20150189843A1 (en) | MAIZE CYTOPLASMIC MALE STERILITY (CMS) S-TYPE RESTORER Rf3 GENE, MOLECULAR MARKERS AND THEIR USE | |
| CN102618510A (zh) | 一种植物雄性育性相关蛋白及其编码基因与应用 | |
| CN106754967A (zh) | 一种水稻粒型基因OsLG1及其编码蛋白质和应用 | |
| WO2022109764A1 (zh) | 一种育性相关基因及其在杂交育种中的应用 | |
| CN102925478B (zh) | 一种水稻雄性育性可控系的构建方法及育种方法 | |
| CN109295071B (zh) | 一种水稻花器官发育调控基因peh1及其编码的蛋白质和应用 | |
| CN114853857B (zh) | 调控玉米花粉育性和花粉耐热性的方法 | |
| CN113943356B (zh) | 蛋白质phd11、其编码基因以及它们在选育玉米雄性不育系中的应用 | |
| CN113754746B (zh) | 水稻雄性育性调控基因、其应用以及利用CRISPR-Cas9调控水稻育性的方法 | |
| CN112457385B (zh) | 一种控制水稻生育期基因ljp1的应用 | |
| CN112301035B (zh) | 高抗水稻白叶枯病基因Xa7及其用途 | |
| CN111304219B (zh) | 一种分离自水稻wz1中的gl1基因及其在增加水稻粒长中的应用 | |
| CN107164390A (zh) | 水稻杂种花粉育性基因座Sc的等位基因Sc‑j和Sc‑i的分离克隆和在育种中的应用 | |
| CN113151295A (zh) | 水稻温敏雄性不育基因OsFMS1及其应用 | |
| CN110846325B (zh) | 一种水稻多花基因mof1及其编码的蛋白质的应用 | |
| CN114656538B (zh) | 蛋白质ms28、其编码基因以及它们在选育玉米雄性不育系中的应用 | |
| CN112080481B (zh) | 穗型相关基因OsFRS5及其应用和表型恢复的方法 | |
| CN117402887B (zh) | 一种玉米雄性育性调控基因ZmMS2085及其突变体与应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Huang Wei Inventor after: Li Yunfei Inventor after: Jin Weiwei Inventor after: Zhao Xiaoming Inventor before: Jin Weiwei Inventor before: Li Yunfei Inventor before: Huang Wei Inventor before: Zhao Xiaoming |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |