CN108794612B - 植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用 - Google Patents
植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用 Download PDFInfo
- Publication number
- CN108794612B CN108794612B CN201810686038.7A CN201810686038A CN108794612B CN 108794612 B CN108794612 B CN 108794612B CN 201810686038 A CN201810686038 A CN 201810686038A CN 108794612 B CN108794612 B CN 108794612B
- Authority
- CN
- China
- Prior art keywords
- fatty acid
- plant
- sequence
- ghbzip67
- protein
- 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.)
- Expired - Fee Related
Links
- 235000014113 dietary fatty acids Nutrition 0.000 title claims abstract description 95
- 229930195729 fatty acid Natural products 0.000 title claims abstract description 95
- 239000000194 fatty acid Substances 0.000 title claims abstract description 95
- 150000004665 fatty acids Chemical class 0.000 title claims abstract description 95
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 75
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 36
- 229920000742 Cotton Polymers 0.000 claims abstract description 18
- 238000009395 breeding Methods 0.000 claims abstract description 5
- 230000001488 breeding effect Effects 0.000 claims abstract description 5
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 6
- 241000196324 Embryophyta Species 0.000 claims description 92
- 230000009261 transgenic effect Effects 0.000 claims description 23
- 241000219195 Arabidopsis thaliana Species 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 18
- 108020004414 DNA Proteins 0.000 claims description 11
- 102000053602 DNA Human genes 0.000 claims description 7
- 230000001965 increasing effect Effects 0.000 claims description 6
- 235000015112 vegetable and seed oil Nutrition 0.000 abstract description 5
- 125000000539 amino acid group Chemical group 0.000 abstract description 3
- 235000013311 vegetables Nutrition 0.000 abstract description 2
- 239000013604 expression vector Substances 0.000 description 17
- 241000219146 Gossypium Species 0.000 description 15
- 238000003259 recombinant expression Methods 0.000 description 14
- 239000013598 vector Substances 0.000 description 12
- 239000003921 oil Substances 0.000 description 11
- 235000019198 oils Nutrition 0.000 description 11
- 230000014759 maintenance of location Effects 0.000 description 10
- 235000012343 cottonseed oil Nutrition 0.000 description 7
- 241000589158 Agrobacterium Species 0.000 description 6
- 241000219194 Arabidopsis Species 0.000 description 5
- 108091026890 Coding region Proteins 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 235000009429 Gossypium barbadense Nutrition 0.000 description 4
- 244000299507 Gossypium hirsutum Species 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 239000000835 fiber Substances 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000009466 transformation Effects 0.000 description 4
- 235000018322 upland cotton Nutrition 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 150000001413 amino acids Chemical group 0.000 description 3
- 108010062796 arginyllysine Proteins 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 229930027917 kanamycin Natural products 0.000 description 3
- 229960000318 kanamycin Drugs 0.000 description 3
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 3
- 229930182823 kanamycin A Natural products 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- NKCZYEDZTKOFBG-GUBZILKMSA-N Gln-Gln-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NKCZYEDZTKOFBG-GUBZILKMSA-N 0.000 description 2
- 235000009438 Gossypium Nutrition 0.000 description 2
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 2
- 240000001140 Mimosa pudica Species 0.000 description 2
- 235000016462 Mimosa pudica Nutrition 0.000 description 2
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 108091081024 Start codon Proteins 0.000 description 2
- 108010028230 Trp-Ser- His-Pro-Gln-Phe-Glu-Lys Proteins 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 108010008355 arginyl-glutamine Proteins 0.000 description 2
- 108010068380 arginylarginine Proteins 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000010170 biological method Methods 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 108010019832 glycyl-asparaginyl-glycine Proteins 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 108010005942 methionylglycine Proteins 0.000 description 2
- 238000000520 microinjection Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000008635 plant growth Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 108010077112 prolyl-proline Proteins 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000005026 transcription initiation Effects 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- CXRCVCURMBFFOL-FXQIFTODSA-N Ala-Ala-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CXRCVCURMBFFOL-FXQIFTODSA-N 0.000 description 1
- QDRGPQWIVZNJQD-CIUDSAMLSA-N Ala-Arg-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O QDRGPQWIVZNJQD-CIUDSAMLSA-N 0.000 description 1
- LWUWMHIOBPTZBA-DCAQKATOSA-N Ala-Arg-Lys Chemical compound NC(=N)NCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CCCCN)C(O)=O LWUWMHIOBPTZBA-DCAQKATOSA-N 0.000 description 1
- SHYYAQLDNVHPFT-DLOVCJGASA-N Ala-Asn-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 SHYYAQLDNVHPFT-DLOVCJGASA-N 0.000 description 1
- CXQODNIBUNQWAS-CIUDSAMLSA-N Ala-Gln-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N CXQODNIBUNQWAS-CIUDSAMLSA-N 0.000 description 1
- NBTGEURICRTMGL-WHFBIAKZSA-N Ala-Gly-Ser Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O NBTGEURICRTMGL-WHFBIAKZSA-N 0.000 description 1
- HOVPGJUNRLMIOZ-CIUDSAMLSA-N Ala-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C)N HOVPGJUNRLMIOZ-CIUDSAMLSA-N 0.000 description 1
- NZGRHTKZFSVPAN-BIIVOSGPSA-N Ala-Ser-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N NZGRHTKZFSVPAN-BIIVOSGPSA-N 0.000 description 1
- 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 1
- ZXKNLCPUNZPFGY-LEWSCRJBSA-N Ala-Tyr-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N2CCC[C@@H]2C(=O)O)N ZXKNLCPUNZPFGY-LEWSCRJBSA-N 0.000 description 1
- GIVWETPOBCRTND-DCAQKATOSA-N Arg-Gln-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O GIVWETPOBCRTND-DCAQKATOSA-N 0.000 description 1
- FFEUXEAKYRCACT-PEDHHIEDSA-N Arg-Ile-Ile Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCCNC(N)=N)[C@@H](C)CC)C(O)=O FFEUXEAKYRCACT-PEDHHIEDSA-N 0.000 description 1
- RIIVUOJDDQXHRV-SRVKXCTJSA-N Arg-Lys-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O RIIVUOJDDQXHRV-SRVKXCTJSA-N 0.000 description 1
- JBIRFLWXWDSDTR-CYDGBPFRSA-N Arg-Met-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCN=C(N)N)N JBIRFLWXWDSDTR-CYDGBPFRSA-N 0.000 description 1
- VENMDXUVHSKEIN-GUBZILKMSA-N Arg-Ser-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O VENMDXUVHSKEIN-GUBZILKMSA-N 0.000 description 1
- SYFHFLGAROUHNT-VEVYYDQMSA-N Arg-Thr-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O SYFHFLGAROUHNT-VEVYYDQMSA-N 0.000 description 1
- RYQSYXFGFOTJDJ-RHYQMDGZSA-N Arg-Thr-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O RYQSYXFGFOTJDJ-RHYQMDGZSA-N 0.000 description 1
- XRNXPIGJPQHCPC-RCWTZXSCSA-N Arg-Thr-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCCNC(N)=N)[C@@H](C)O)C(O)=O XRNXPIGJPQHCPC-RCWTZXSCSA-N 0.000 description 1
- MFFOYNGMOYFPBD-DCAQKATOSA-N Asn-Arg-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O MFFOYNGMOYFPBD-DCAQKATOSA-N 0.000 description 1
- RAQMSGVCGSJKCL-FOHZUACHSA-N Asn-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(N)=O RAQMSGVCGSJKCL-FOHZUACHSA-N 0.000 description 1
- ZYPWIUFLYMQZBS-SRVKXCTJSA-N Asn-Lys-Lys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N ZYPWIUFLYMQZBS-SRVKXCTJSA-N 0.000 description 1
- SNYCNNPOFYBCEK-ZLUOBGJFSA-N Asn-Ser-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O SNYCNNPOFYBCEK-ZLUOBGJFSA-N 0.000 description 1
- CBWCQCANJSGUOH-ZKWXMUAHSA-N Asn-Val-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O CBWCQCANJSGUOH-ZKWXMUAHSA-N 0.000 description 1
- WLKVEEODTPQPLI-ACZMJKKPSA-N Asp-Gln-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O WLKVEEODTPQPLI-ACZMJKKPSA-N 0.000 description 1
- VILLWIDTHYPSLC-PEFMBERDSA-N Asp-Glu-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VILLWIDTHYPSLC-PEFMBERDSA-N 0.000 description 1
- OVPHVTCDVYYTHN-AVGNSLFASA-N Asp-Glu-Phe Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 OVPHVTCDVYYTHN-AVGNSLFASA-N 0.000 description 1
- POTCZYQVVNXUIG-BQBZGAKWSA-N Asp-Gly-Pro Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O POTCZYQVVNXUIG-BQBZGAKWSA-N 0.000 description 1
- CTWCFPWFIGRAEP-CIUDSAMLSA-N Asp-Lys-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O CTWCFPWFIGRAEP-CIUDSAMLSA-N 0.000 description 1
- FQHBAQLBIXLWAG-DCAQKATOSA-N Asp-Lys-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(=O)O)N FQHBAQLBIXLWAG-DCAQKATOSA-N 0.000 description 1
- LTCKTLYKRMCFOC-KKUMJFAQSA-N Asp-Phe-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O LTCKTLYKRMCFOC-KKUMJFAQSA-N 0.000 description 1
- 241000219193 Brassicaceae Species 0.000 description 1
- 241000722721 Capparis Species 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- XZWYTXMRWQJBGX-VXBMVYAYSA-N FLAG peptide Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(O)=O)CC1=CC=C(O)C=C1 XZWYTXMRWQJBGX-VXBMVYAYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- OYTPNWYZORARHL-XHNCKOQMSA-N Gln-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N OYTPNWYZORARHL-XHNCKOQMSA-N 0.000 description 1
- KZKBJEUWNMQTLV-XDTLVQLUSA-N Gln-Ala-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KZKBJEUWNMQTLV-XDTLVQLUSA-N 0.000 description 1
- RKAQZCDMSUQTSS-FXQIFTODSA-N Gln-Asp-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N RKAQZCDMSUQTSS-FXQIFTODSA-N 0.000 description 1
- MADFVRSKEIEZHZ-DCAQKATOSA-N Gln-Gln-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)N)N MADFVRSKEIEZHZ-DCAQKATOSA-N 0.000 description 1
- NPTGGVQJYRSMCM-GLLZPBPUSA-N Gln-Gln-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O NPTGGVQJYRSMCM-GLLZPBPUSA-N 0.000 description 1
- VOLVNCMGXWDDQY-LPEHRKFASA-N Gln-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCC(=O)N)N)C(=O)O VOLVNCMGXWDDQY-LPEHRKFASA-N 0.000 description 1
- FFVXLVGUJBCKRX-UKJIMTQDSA-N Gln-Ile-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CCC(=O)N)N FFVXLVGUJBCKRX-UKJIMTQDSA-N 0.000 description 1
- IULKWYSYZSURJK-AVGNSLFASA-N Gln-Leu-Lys Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O IULKWYSYZSURJK-AVGNSLFASA-N 0.000 description 1
- IOFDDSNZJDIGPB-GVXVVHGQSA-N Gln-Leu-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O IOFDDSNZJDIGPB-GVXVVHGQSA-N 0.000 description 1
- GURIQZQSTBBHRV-SRVKXCTJSA-N Gln-Lys-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O GURIQZQSTBBHRV-SRVKXCTJSA-N 0.000 description 1
- KPNWAJMEMRCLAL-GUBZILKMSA-N Gln-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N KPNWAJMEMRCLAL-GUBZILKMSA-N 0.000 description 1
- FHPXTPQBODWBIY-CIUDSAMLSA-N Glu-Ala-Arg Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FHPXTPQBODWBIY-CIUDSAMLSA-N 0.000 description 1
- FYBSCGZLICNOBA-XQXXSGGOSA-N Glu-Ala-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FYBSCGZLICNOBA-XQXXSGGOSA-N 0.000 description 1
- AKJRHDMTEJXTPV-ACZMJKKPSA-N Glu-Asn-Ala Chemical compound C[C@H](NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCC(O)=O)C(O)=O AKJRHDMTEJXTPV-ACZMJKKPSA-N 0.000 description 1
- YYOBUPFZLKQUAX-FXQIFTODSA-N Glu-Asn-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O YYOBUPFZLKQUAX-FXQIFTODSA-N 0.000 description 1
- CKRUHITYRFNUKW-WDSKDSINSA-N Glu-Asn-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O CKRUHITYRFNUKW-WDSKDSINSA-N 0.000 description 1
- QQLBPVKLJBAXBS-FXQIFTODSA-N Glu-Glu-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O QQLBPVKLJBAXBS-FXQIFTODSA-N 0.000 description 1
- UGSVSNXPJJDJKL-SDDRHHMPSA-N Glu-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)O)N UGSVSNXPJJDJKL-SDDRHHMPSA-N 0.000 description 1
- NJCALAAIGREHDR-WDCWCFNPSA-N Glu-Leu-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O NJCALAAIGREHDR-WDCWCFNPSA-N 0.000 description 1
- BPLNJYHNAJVLRT-ACZMJKKPSA-N Glu-Ser-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O BPLNJYHNAJVLRT-ACZMJKKPSA-N 0.000 description 1
- GGEJHJIXRBTJPD-BYPYZUCNSA-N Gly-Asn-Gly Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O GGEJHJIXRBTJPD-BYPYZUCNSA-N 0.000 description 1
- QSVCIFZPGLOZGH-WDSKDSINSA-N Gly-Glu-Ser Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O QSVCIFZPGLOZGH-WDSKDSINSA-N 0.000 description 1
- QPTNELDXWKRIFX-YFKPBYRVSA-N Gly-Gly-Gln Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCC(N)=O QPTNELDXWKRIFX-YFKPBYRVSA-N 0.000 description 1
- ADZGCWWDPFDHCY-ZETCQYMHSA-N Gly-His-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 ADZGCWWDPFDHCY-ZETCQYMHSA-N 0.000 description 1
- CCBIBMKQNXHNIN-ZETCQYMHSA-N Gly-Leu-Gly Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O CCBIBMKQNXHNIN-ZETCQYMHSA-N 0.000 description 1
- HHRODZSXDXMUHS-LURJTMIESA-N Gly-Met-Gly Chemical compound CSCC[C@H](NC(=O)C[NH3+])C(=O)NCC([O-])=O HHRODZSXDXMUHS-LURJTMIESA-N 0.000 description 1
- KSOBNUBCYHGUKH-UWVGGRQHSA-N Gly-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN KSOBNUBCYHGUKH-UWVGGRQHSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- OSZUPUINVNPCOE-SDDRHHMPSA-N His-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O OSZUPUINVNPCOE-SDDRHHMPSA-N 0.000 description 1
- 108010093488 His-His-His-His-His-His Proteins 0.000 description 1
- MCGOGXFMKHPMSQ-AVGNSLFASA-N His-Val-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CN=CN1 MCGOGXFMKHPMSQ-AVGNSLFASA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- VSZALHITQINTGC-GHCJXIJMSA-N Ile-Ala-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CC(=O)O)C(=O)O)N VSZALHITQINTGC-GHCJXIJMSA-N 0.000 description 1
- LKACSKJPTFSBHR-MNXVOIDGSA-N Ile-Gln-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N LKACSKJPTFSBHR-MNXVOIDGSA-N 0.000 description 1
- UIEZQYNXCYHMQS-BJDJZHNGSA-N Ile-Lys-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)O)N UIEZQYNXCYHMQS-BJDJZHNGSA-N 0.000 description 1
- FGBRXCZYVRFNKQ-MXAVVETBSA-N Ile-Phe-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)O)N FGBRXCZYVRFNKQ-MXAVVETBSA-N 0.000 description 1
- VEPIBPGLTLPBDW-URLPEUOOSA-N Ile-Phe-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N VEPIBPGLTLPBDW-URLPEUOOSA-N 0.000 description 1
- KTNGVMMGIQWIDV-OSUNSFLBSA-N Ile-Pro-Thr Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O KTNGVMMGIQWIDV-OSUNSFLBSA-N 0.000 description 1
- 108020005350 Initiator Codon Proteins 0.000 description 1
- CZCSUZMIRKFFFA-CIUDSAMLSA-N Leu-Ala-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O CZCSUZMIRKFFFA-CIUDSAMLSA-N 0.000 description 1
- RFUBXQQFJFGJFV-GUBZILKMSA-N Leu-Asn-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O RFUBXQQFJFGJFV-GUBZILKMSA-N 0.000 description 1
- QVFGXCVIXXBFHO-AVGNSLFASA-N Leu-Glu-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O QVFGXCVIXXBFHO-AVGNSLFASA-N 0.000 description 1
- OGUUKPXUTHOIAV-SDDRHHMPSA-N Leu-Glu-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N OGUUKPXUTHOIAV-SDDRHHMPSA-N 0.000 description 1
- ZGUMORRUBUCXEH-AVGNSLFASA-N Leu-Lys-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZGUMORRUBUCXEH-AVGNSLFASA-N 0.000 description 1
- QWWPYKKLXWOITQ-VOAKCMCISA-N Leu-Thr-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(C)C QWWPYKKLXWOITQ-VOAKCMCISA-N 0.000 description 1
- YWFZWQKWNDOWPA-XIRDDKMYSA-N Leu-Trp-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(O)=O YWFZWQKWNDOWPA-XIRDDKMYSA-N 0.000 description 1
- MPOHDJKRBLVGCT-CIUDSAMLSA-N Lys-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCCN)N MPOHDJKRBLVGCT-CIUDSAMLSA-N 0.000 description 1
- DNEJSAIMVANNPA-DCAQKATOSA-N Lys-Asn-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O DNEJSAIMVANNPA-DCAQKATOSA-N 0.000 description 1
- HWMZUBUEOYAQSC-DCAQKATOSA-N Lys-Gln-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O HWMZUBUEOYAQSC-DCAQKATOSA-N 0.000 description 1
- VSRXPEHZMHSFKU-IUCAKERBSA-N Lys-Gln-Gly Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O VSRXPEHZMHSFKU-IUCAKERBSA-N 0.000 description 1
- QZONCCHVHCOBSK-YUMQZZPRSA-N Lys-Gly-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O QZONCCHVHCOBSK-YUMQZZPRSA-N 0.000 description 1
- 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 1
- 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 1
- PLDJDCJLRCYPJB-VOAKCMCISA-N Lys-Lys-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PLDJDCJLRCYPJB-VOAKCMCISA-N 0.000 description 1
- QQPSCXKFDSORFT-IHRRRGAJSA-N Lys-Lys-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCCN QQPSCXKFDSORFT-IHRRRGAJSA-N 0.000 description 1
- IKXQOBUBZSOWDY-AVGNSLFASA-N Lys-Val-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CCCCN)N IKXQOBUBZSOWDY-AVGNSLFASA-N 0.000 description 1
- DBOMZJOESVYERT-GUBZILKMSA-N Met-Asn-Met Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCSC)C(=O)O)N DBOMZJOESVYERT-GUBZILKMSA-N 0.000 description 1
- XDGFFEZAZHRZFR-RHYQMDGZSA-N Met-Leu-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XDGFFEZAZHRZFR-RHYQMDGZSA-N 0.000 description 1
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 1
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- ZLGQEBCCANLYRA-RYUDHWBXSA-N Phe-Gly-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O ZLGQEBCCANLYRA-RYUDHWBXSA-N 0.000 description 1
- BIYWZVCPZIFGPY-QWRGUYRKSA-N Phe-Gly-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CO)C(O)=O BIYWZVCPZIFGPY-QWRGUYRKSA-N 0.000 description 1
- MMJJFXWMCMJMQA-STQMWFEESA-N Phe-Pro-Gly Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)NCC(O)=O)C1=CC=CC=C1 MMJJFXWMCMJMQA-STQMWFEESA-N 0.000 description 1
- CYQQWUPHIZVCNY-GUBZILKMSA-N Pro-Arg-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O CYQQWUPHIZVCNY-GUBZILKMSA-N 0.000 description 1
- LSIWVWRUTKPXDS-DCAQKATOSA-N Pro-Gln-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O LSIWVWRUTKPXDS-DCAQKATOSA-N 0.000 description 1
- 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 1
- HJSCRFZVGXAGNG-SRVKXCTJSA-N Pro-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1 HJSCRFZVGXAGNG-SRVKXCTJSA-N 0.000 description 1
- VPEVBAUSTBWQHN-NHCYSSNCSA-N Pro-Glu-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O VPEVBAUSTBWQHN-NHCYSSNCSA-N 0.000 description 1
- NFLNBHLMLYALOO-DCAQKATOSA-N Pro-Leu-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@@H]1CCCN1 NFLNBHLMLYALOO-DCAQKATOSA-N 0.000 description 1
- OWQXAJQZLWHPBH-FXQIFTODSA-N Pro-Ser-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O OWQXAJQZLWHPBH-FXQIFTODSA-N 0.000 description 1
- FDMCIBSQRKFSTJ-RHYQMDGZSA-N Pro-Thr-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O FDMCIBSQRKFSTJ-RHYQMDGZSA-N 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- YQHZVYJAGWMHES-ZLUOBGJFSA-N Ser-Ala-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YQHZVYJAGWMHES-ZLUOBGJFSA-N 0.000 description 1
- CRZRTKAVUUGKEQ-ACZMJKKPSA-N Ser-Gln-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O CRZRTKAVUUGKEQ-ACZMJKKPSA-N 0.000 description 1
- IXUGADGDCQDLSA-FXQIFTODSA-N Ser-Gln-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CO)N IXUGADGDCQDLSA-FXQIFTODSA-N 0.000 description 1
- WSTIOCFMWXNOCX-YUMQZZPRSA-N Ser-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CO)N WSTIOCFMWXNOCX-YUMQZZPRSA-N 0.000 description 1
- CJINPXGSKSZQNE-KBIXCLLPSA-N Ser-Ile-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(O)=O CJINPXGSKSZQNE-KBIXCLLPSA-N 0.000 description 1
- KCNSGAMPBPYUAI-CIUDSAMLSA-N Ser-Leu-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O KCNSGAMPBPYUAI-CIUDSAMLSA-N 0.000 description 1
- XNCUYZKGQOCOQH-YUMQZZPRSA-N Ser-Leu-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O XNCUYZKGQOCOQH-YUMQZZPRSA-N 0.000 description 1
- MQUZANJDFOQOBX-SRVKXCTJSA-N Ser-Phe-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O MQUZANJDFOQOBX-SRVKXCTJSA-N 0.000 description 1
- JGUWRQWULDWNCM-FXQIFTODSA-N Ser-Val-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O JGUWRQWULDWNCM-FXQIFTODSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- VTVVYQOXJCZVEB-WDCWCFNPSA-N Thr-Leu-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O VTVVYQOXJCZVEB-WDCWCFNPSA-N 0.000 description 1
- UXUAZXWKIGPUCH-RCWTZXSCSA-N Thr-Met-Met Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCSC)C(O)=O UXUAZXWKIGPUCH-RCWTZXSCSA-N 0.000 description 1
- AHERARIZBPOMNU-KATARQTJSA-N Thr-Ser-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O AHERARIZBPOMNU-KATARQTJSA-N 0.000 description 1
- KPMIQCXJDVKWKO-IFFSRLJSSA-N Thr-Val-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KPMIQCXJDVKWKO-IFFSRLJSSA-N 0.000 description 1
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 1
- YTVJTXJTNRWJCR-JBACZVJFSA-N Trp-Phe-Glu Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N YTVJTXJTNRWJCR-JBACZVJFSA-N 0.000 description 1
- VCXWRWYFJLXITF-AUTRQRHGSA-N Tyr-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 VCXWRWYFJLXITF-AUTRQRHGSA-N 0.000 description 1
- OLWFDNLLBWQWCP-STQMWFEESA-N Tyr-Gly-Met Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O OLWFDNLLBWQWCP-STQMWFEESA-N 0.000 description 1
- 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 1
- BRPKEERLGYNCNC-NHCYSSNCSA-N Val-Glu-Arg Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N BRPKEERLGYNCNC-NHCYSSNCSA-N 0.000 description 1
- WNZSAUMKZQXHNC-UKJIMTQDSA-N Val-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N WNZSAUMKZQXHNC-UKJIMTQDSA-N 0.000 description 1
- AEMPCGRFEZTWIF-IHRRRGAJSA-N Val-Leu-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O AEMPCGRFEZTWIF-IHRRRGAJSA-N 0.000 description 1
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 1
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000002551 biofuel Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000000546 chi-square test Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000002385 cottonseed oil Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- 108010048994 glycyl-tyrosyl-alanine Proteins 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 108010051673 leucyl-glycyl-phenylalanine Proteins 0.000 description 1
- 108010009298 lysylglutamic acid Proteins 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000003147 molecular marker Substances 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000013421 nuclear magnetic resonance imaging Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 108010073101 phenylalanylleucine Proteins 0.000 description 1
- 238000003976 plant breeding Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- 108010053725 prolylvaline Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 1
- 229960001225 rifampicin Drugs 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 108010071207 serylmethionine Proteins 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000009941 weaving Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—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
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
- C12N15/8247—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified lipid metabolism, e.g. seed oil composition
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Nutrition Science (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Botany (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
本发明公开了一种植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用。本发明提供的蛋白质，是如下(a1)或(a2)：(a1)由序列表中序列1所示的氨基酸序列组成的蛋白质；(a2)将序列表中序列1所示的氨基酸序列经过一个或几个氨基酸残基的取代和/或缺失和/或添加且与植物种子油分相关的由序列1衍生的蛋白质。本发明提供了GhbZIP67蛋白及其编码基因，可以调控植物种子脂肪酸含量。本发明对于培育棉花新品种具有重要的意义，适合于推广应用。
Description
技术领域
本发明涉及一种植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用。
背景技术
棉花是重要的纤维作物，也是重要的油料作物。在陆地棉的棉仁中，油分约占25％-40％，其含油量与大豆相当，是植物油的重要来源，棉花在世界油料作物中排名第六。研究者对于陆地棉的研究主要集中于纤维品质、产量性状以及抗性性状，对棉籽含油量性状研究较少。陆地棉种植面积广泛，在生产上每产生1千克纤维的同时产生约1.65千克的棉籽，世界上每年可生产大量棉籽，陆地棉棉籽可以作为重要的油料来源。随着人口的增长对食用植物油的需求越来越大，以及生物燃料的推广应用，棉花的副产品棉籽受到越来越多的关注，提高棉籽含油量具有重要的意义。
棉籽油分相关基因的定位工作进展缓慢，国内外研究者，对于棉花QTLs的研究，主要集中于纤维品质相关性状的QTLs、产量相关性状的QTLs等，对棉籽含油量的研究相对较少，并且由于分子标记密度的限制，目前通过正向遗传学图位克隆的方法克隆油分相关基因还非常困难。
发明内容
本发明的目的是提供一种植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用。
本发明提供的蛋白质，获自棉花，命名为GhbZIP67蛋白，是如下(a1)或(a2)：
(a1)由序列表中序列1所示的氨基酸序列组成的蛋白质；
(a2)将序列表中序列1所示的氨基酸序列经过一个或几个氨基酸残基的取代和/或缺失和/或添加且与植物种子油分相关的由序列1衍生的蛋白质。
为了使(a1)中的GhbZIP67蛋白便于纯化和检测，可在由序列表中序列1所示的氨基酸序列组成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
| 标签 | 残基 | 序列 |
| Poly-Arg | 5-6(通常为5个) | RRRRR |
| Poly-His | 2-10(通常为6个) | HHHHHH |
| FLAG | 8 | DYKDDDDK |
| Strep-tag II | 8 | WSHPQFEK |
| c-myc | 10 | EQKLISEEDL |
上述(a2)中的GhbZIP67蛋白可人工合成，也可先合成其编码基因，再进行生物表达得到。上述(a2)中的GhbZIP67蛋白的编码基因可通过将序列表中序列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。
编码所述GhbZIP67蛋白的基因(GhbZIP67基因)也属于本发明的保护范围。
所述基因为如下(b1)-(b3)中任一所述的DNA分子：
(b1)编码区如序列表中序列2所示的DNA分子；
(b2)在严格条件下与(b1)限定的DNA序列杂交且编码与植物种子油分相关的蛋白的DNA分子；
(b3)与(b1)或(b2)限定的DNA序列具有90％以上同源性且编码与植物种子油分相关的的蛋白的DNA分子。
上述严格条件可为用0.1×SSPE(或0.1×SSC)，0.1％SDS的溶液，在DNA或者RNA杂交实验中65℃下杂交并洗膜。
含有GhbZIP67基因的重组表达载体、表达盒、转基因细胞系或重组菌也属于本发明的保护范围。
可用现有的植物表达载体构建含有GhbZIP67基因的重组表达载体。所述植物表达载体包括双元农杆菌载体和可用于植物微弹轰击的载体等。使用GhbZIP67基因构建重组表达载体时，可在其转录起始核苷酸前加上任何一种增强型、组成型、组织特异型或诱导型启动子，它们可单独使用或与其它的植物启动子结合使用；此外，使用GhbZIP67基因构建重组表达载体时，还可使用增强子，包括翻译增强子或转录增强子，这些增强子区域可以是ATG起始密码子或邻接区域起始密码子等，但必需与编码序列的阅读框相同，以保证整个序列的正确翻译。所述翻译控制信号和起始密码子的来源是广泛的，可以是天然的，也可以是合成的。翻译起始区域可以来自转录起始区域或结构基因。为了便于对转基因植物细胞或植物进行鉴定及筛选，可对所用植物表达载体进行加工，如加入在植物中表达可产生颜色变化的酶或发光化合物的基因、具有抗性的抗生素标记物或是抗化学试剂标记基因等。
所述重组表达载体具体可为将pBI121载体的BamHI和SacI酶切位点间的片段替换为序列表的序列2自5’端第1-1176位所示的DNA分子得到的重组表达载体。
本发明还保护GhbZIP67蛋白或GhbZIP67基因调控植物种子脂肪酸含量中的应用。
所述脂肪酸具体可为18:0脂肪酸、18:1脂肪酸、20:0脂肪酸、20:1脂肪酸、16:0脂肪酸、16:1脂肪酸、18:3脂肪酸或22:1脂肪酸。
本发明还保护一种培育转基因植物的方法(方法甲)，包括如下步骤：将GhbZIP67基因导入出发植物，得到种子脂肪酸含量提高的转基因植物；所述脂肪酸为18:0脂肪酸、18:1脂肪酸、20:0脂肪酸或20:1脂肪酸。
所述方法中，所述GhbZIP67基因可以通过以上任一所述重组表达载体导入目的植物。所述重组表达载体可通过Ti质粒、Ri质粒、植物病毒载体、直接DNA转化、显微注射、电导、农杆菌介导等常规生物学方法转化到植物细胞或组织中。
本发明还保护一种提高植物种子脂肪酸含量的方法，包括如下步骤：提高出发植物中GhbZIP67蛋白的活性和/或表达量，得到种子脂肪酸含量提高的植物；所述脂肪酸为18:0脂肪酸、18:1脂肪酸、20:0脂肪酸或20:1脂肪酸。
本发明还保护一种培育转基因植物的方法，包括如下步骤：将GhbZIP67基因导入出发植物，得到种子脂肪酸含量降低的转基因植物；所述脂肪酸为16:0脂肪酸、16:1脂肪酸、18:3脂肪酸或22:1脂肪酸。
所述方法中，所述GhbZIP67基因可以通过以上任一所述重组表达载体导入目的植物。所述重组表达载体可通过Ti质粒、Ri质粒、植物病毒载体、直接DNA转化、显微注射、电导、农杆菌介导等常规生物学方法转化到植物细胞或组织中。
本发明还保护一种降低植物种子脂肪酸含量的方法，包括如下步骤：提高出发植物中GhbZIP67蛋白的活性和/或表达量，得到种子脂肪酸含量降低的植物；所述脂肪酸为16:0脂肪酸、16:1脂肪酸、18:3脂肪酸或22:1脂肪酸。
本发明还保护GhbZIP67蛋白，或，GhbZIP67基因，或，以上任一所述的方法在植物育种中的应用。
所述育种的目的是选育种子脂肪酸含量升高或降低的植物。所述脂肪酸具体可为18:0脂肪酸、18:1脂肪酸、20:0脂肪酸、20:1脂肪酸、16:0脂肪酸、16:1脂肪酸、18:3脂肪酸或22:1脂肪酸。
以上任一所述植物为双子叶植物或单子叶植物。所述双子叶植物可为山柑目植物。所述山柑目植物可为十字花科植物。所述十字花科植物可为南芥族植物。所述南芥族植物可为拟南芥属植物。所述所述拟南芥属植物具体可为拟南芥，例如哥伦比亚生态型拟南芥。
本发明提供了GhbZIP67蛋白及其编码基因，可以调控植物种子脂肪酸含量。本发明对于培育棉花新品种具有重要的意义，适合于推广应用。
附图说明
图1为转基因植株的qRT-PCR检测结果。
图2为转基因植株的总脂肪含量检测结果。
具体实施方式
以下的实施例便于更好地理解本发明，但并不限定本发明。下述实施例中的实验方法，如无特殊说明，均为常规方法。下述实施例中所用的试验材料，如无特殊说明，均为自常规生化试剂商店购买得到的。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。
pBI121载体：参考文献：Chen,P.Y.,Wang,C.K.,Soong,S.C.,&To,K.Y.(2003).Complete sequence of the binary vector pBI121 and its application in cloningT-DNA insertion from transgenic plants.Molecular breeding,11(4),287-293.；公众可以从中国农业科学院棉花研究所获得。
农杆菌GV3101：郑州尚医生物科技有限公司(华越洋品牌)。
拟南芥Col-0：参考文献：梅雄,李振江,张慧,等.3种接种方法对十字花科黑腐病菌Xcc8004菌株在拟南芥Col-0上致病力的影响[J].基因组学与应用生物学,2011,30(4):365-370.；公众可以从中国农业科学院棉花研究所获得。
实施例1、GhBZIP67蛋白及其编码基因的获得
提取棉花陆海回交近交系3008不同组织器官材料的总RNA，并反转录为cDNA。经过大量序列分析、表达量分析与功能验证，从cDNA中发现了一个DNA编码序列，如序列表的序列2所示，其编码的蛋白质如序列表的序列1所示。
将序列表的序列1所示的蛋白质命名为GhBZIP67蛋白。将编码GhBZIP67蛋白的基因命名为GhBZIP67基因。
实施例2、GhBZIP67蛋白及其编码基因的功能验证
一、转基因植株的获得
1、重组表达载体的构建：将pBI121载体的BamHI和SacI酶切位点间的片段替换为序列表的序列2自5’端第1-1176位所示的DNA分子，得到重组表达载体pBI121::GhBZIP67(已经测序验证)。
2、将步骤1得到的重组表达载体pBI121::GhBZIP67导入农杆菌GV3101，得到重组菌GV3101::GhBZIP67。
3、将步骤2得到的重组菌GV3101::GhBZIP67通过蘸花法转入拟南芥Col-0中，具体转化步骤如下：
(1)将步骤2得到的重组菌GV3101::GhBZIP67接种于含有100mg/L利福平、25mg/L庆大和50mg/L卡纳霉素的YEB液体培养基中，28℃、200rpm培养至菌液OD600nm为0.6。
(2)完成步骤(1)后，将培养体系离心，收集菌体沉淀，采用含有5g/100mL蔗糖和20μL/100mL 1 Silvet的/2MS液体培养基重悬菌体沉淀并调整菌浓度至菌液OD600nm为2.0。
(3)取步骤(2)得到的菌液，采用蘸花法转化拟南芥(方法参照文献：Zhang,X.,Henriques,R.,Lin,S.S.,Niu,Q.W.,and Chua,N.H.(2006).Agrobacterium-mediatedtransformation of Arabidopsis thaliana using the floral dip method.Nat Protoc1,641-646.doi:10.1038/nprot.2006.97)，转化后的拟南芥避光培养24h后，恢复到正常条件(22℃、光照16h/18℃、黑暗8h，湿度60％)培养。
每一周重复上述步骤转化一次，一共转化3次，直至收获T0代转基因种子。
4、取步骤3得到的T0代转基因种子，消毒后在4℃避光放置3天，然后平铺于含有50mg/L卡纳霉素的MS固体培养基上，在正常条件(22℃、光照16h/18℃、黑暗8h，湿度60％)下培养10天，筛选得到T1代抗性植株(抗性植株表型：植株正常生长，叶片深绿，根系发育正常；敏感植株表型：植株停止生长，叶片黄花，无根或根系非常短)。
5、将步骤4得到的T1代抗性植株移栽到营养土中并继续在正常条件(22℃、光照16h/18℃、黑暗8h，湿度60％)下培养，得到T2代植株，按照步骤4中的标准筛选得到T2代抗性植株，如果T2代植株中满足抗性植株：敏感植株＝3:1的条件(符合卡方测验)，其T1代及自交后代为单拷贝插入株。
6、取步骤5得到的单拷贝插入株的T2代卡纳霉素抗性植株自交并收获T3代植株，按照步骤4中的标准筛选得到T3代抗性植株，对于某一T2代植株来说，如果其抽样检测的T3代植株均为抗性植株，则认为该T2代植株为纯合的转基因植株，该T2代植株及其自交后代为一个纯合的转基因株系。
二、转空载体植株的获得
采用pBI121载体替代重组表达载体pBI121::GhBZIP67，按照步骤3-6进行操作，得到转空载体株系。
三、转基因植株的qRT-PCR检测
待测植株：拟南芥Col-0(WT)、转基因株系(L1、L2、L3)的T3代植株、转空载体株系T3代植株。
1、提取待测植株果荚的总RNA，并反转录为cDNA。
2、以步骤1得到的cDNA为模板，采用引物qRT-GhBZIP67-F和引物qRT-GhBZIP67-R组成的引物对进行荧光定量PCR，检测待测植株中GhBZIP67基因的表达情况；采用引物Ghhiston3-F和引物Ghhiston3-R组成的引物对检测内参基因18S。
qRT-GhBZIP67-F：ATTGTTCCCAGCAACTTGG(5’-3’)；
qRT-GhBZIP67-R：TGTATCACGACGGGCTCTAAC(5’-3’)；
Ghhiston3-F：GAAGCCTCATCGATACCGTC(5’-3’)；
Ghhiston3-R：CTACCACTACCATCATGGC(5’-3’)。
结果如图1所示。结果表明，相对于拟南芥Col-0(WT)，转基因株系(L1、L2、L3)中GhBZIP67基因都正常表达。转空载体株系中GhBZIP67基因的表达情况与拟南芥Col-0(WT)相同。
四、功能鉴定
待测植株：拟南芥Col-0(WT)、转基因株系(L1、L2、L3)的T3代植株、转空载体株系T3代植株。
1、总脂肪含量检测
采用核磁共振成像分析仪(Niumag corporation，NMI20-Analyst)并按照操作方法对待测植株种子进行总脂肪含量测定。
结果如图2所示。结果表明，转基因株系L1，L2和L3三个转基因株系油分含量未发生明显改变。转空载体株系油分含量与拟南芥(WT)无差异。
2、脂肪酸成分含量检测
将待测植株种子用研钵研磨成粉末状，取10mg粉末放入5mL具塞离心管中，然后加入200μL的90～120℃的石油醚，在100W超声波45℃的条件下浸提1.5h；然后加入100μL的0.5mol/L的KOH－CH3OH溶液，高速振荡甲酯化；然后加入5μL饱和NaCl水溶液，4000rpm离心10min后，将上清转入1.5mL进样瓶用Agilent7890A气相色谱仪进行脂肪酸成分测定。
色谱柱:DB－23毛细管色谱柱(60m×0.25mm×0.25μm)；柱温：程序升温:100℃保持1min，以25℃/min速率升温到175℃，再以4℃/min的速率升温到230℃，保持5min；FID检测器温度280℃；进样口温度250℃；载气为高纯氮气(纯度99.999％)，流速30mL/min；氢气流速40mL/min；空气流速450mL/min；进样量1.0μL；分流比30：1。
12.106保留时间对应的峰为棕榈酸(C16:0)；14.357保留时间对应的峰为硬脂酸(C18:0)。脂肪酸16:0保留时间10.306，脂肪酸16:1保留时间10.615,脂肪酸18:0保留时间12.284,脂肪酸18:1保留时间12.593和12.676,脂肪酸18:3保留时间13.936,脂肪酸20:0保留时间14.605,脂肪酸20:1保留时间14.970和15.070,脂肪酸22:1保留时间17.528。
结果如表2所示。结果表明，转基因株系相对于拟南芥Col-0(WT)，16:0、16:1、18:3和22:1脂肪酸成分降低，18:0、18:1、20:0和20:1脂肪酸成分升高。转空载体株系脂肪酸成分与拟南芥(WT)无差异。
表2脂肪酸成分测定结果
上述结果说明：本发明的GhbZIP67基因具有调节脂肪酸成分的功能。
序列表
<110> 中国农业科学院棉花研究所
<120> 植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 391
<212> PRT
<213> 棉花(Gossypium spp)
<400> 1
Met Ala Leu Phe Ala Ser Glu Ser Val Ala Tyr Gly Gly Ile Lys Ala
1 5 10 15
Ala Ser Pro Ser Gln Ala Pro Arg Ser Pro Gln Gln Gln Gln Arg Gln
20 25 30
Glu Leu Thr Gln Asp Gln Ser Val Ser Ser Leu Asn Lys Gln Gly Ser
35 40 45
Ile Phe Ser Leu Thr Leu Asp Glu Ile Gln Leu Lys Ser Gly Lys Ala
50 55 60
Phe Gly Ser Met Asn Met Asp Glu Phe Leu Ala Asn Leu Trp Asn Val
65 70 75 80
Glu Glu Asn Gln Ala Pro Pro Gln Leu Asp Gln Asn Glu Ala Arg Asp
85 90 95
Asp Lys Asp Lys Gly Asn Gly Gly Gln Pro Thr Leu Ala Arg Gln Gly
100 105 110
Ser Phe Ser Ile Pro Thr Pro Leu Cys Lys Lys Thr Val Asp Glu Val
115 120 125
Trp Phe Glu Ile Gln Lys Glu Leu Pro Gln Gln Arg Lys Ala Asn Asn
130 135 140
Ile Ala Asp His Glu Pro Pro Gln Arg Gln Gln Thr Phe Gly Glu Met
145 150 155 160
Thr Leu Glu Asp Phe Leu Ile Lys Ala Gly Val Val Gln Glu Pro Ser
165 170 175
Ser Ala Ser Ser Gln Gln Lys Lys Val Ala Ser Thr Arg Thr Asn Gly
180 185 190
Thr Ser Leu Glu Ala Thr Tyr Gly Met Gly His Gly Ala Gly Ser Ser
195 200 205
Gln Gln Lys Met Leu Thr Ser Ile Gln Asn Ser Ser Ala Ser Leu Asp
210 215 220
Ala Asn Phe Gly Met Gly His Val Met Gly Leu Gly Phe Pro Gly His
225 230 235 240
Gln Ile Val Pro Ser Asn Leu Ala Ala Pro Gly Asn Gly Tyr Ala Ala
245 250 255
Ala Tyr Pro Ile Phe Thr Gln Ser Lys Thr Met Met Gly Glu Ser Ser
260 265 270
Asn Val Ala Glu Asn Gly Asn Gly Thr Asn Arg Leu Leu Glu Pro Val
275 280 285
Val Ile Gln Asn Lys Lys Arg Ile Ile Asp Gly Pro Pro Glu Val Val
290 295 300
Val Glu Arg Arg Gln Arg Arg Met Ile Lys Asn Arg Glu Ser Ala Ala
305 310 315 320
Arg Ser Arg Ala Arg Lys Gln Ala Tyr Thr Val Glu Leu Glu Leu Glu
325 330 335
Leu Asn Gln Leu Lys Gln Glu Asn Ala Lys Leu Lys Gln Leu Val Glu
340 345 350
Glu Asn Glu Gln Lys Arg Lys Gln Glu Val Leu Lys Arg Lys Gln Ser
355 360 365
Lys Leu Pro Ala Gln Arg Lys Val Val Asp Lys Met Arg Thr Leu Arg
370 375 380
Arg Thr Val Ser Leu Gly Trp
385 390
<210> 2
<211> 1176
<212> DNA
<213> 棉花(Gossypium spp)
<400> 2
atggcacttt ttgcatcaga aagtgttgcc tatggtggca tcaaggcagc gtcaccatcg 60
caagcaccgc gatcgccgca gcagcaacaa cgccaggaat taacacagga tcaatctgtt 120
tcttccttga acaaacaggg ttcgattttc tccctcaccc tcgacgagat tcaactgaag 180
agcgggaagg cttttgggtc catgaacatg gatgaattcc ttgccaactt gtggaatgtt 240
gaggaaaacc aagctccacc acagctcgac cagaacgagg ctcgcgatga caaagacaag 300
ggcaacggag gtcaaccaac cctggcccga caaggctctt tctccatccc caccccactt 360
tgcaagaaaa ctgtggatga ggtatggttt gagattcaga aagagctacc gcagcagcgg 420
aaggctaaca acatcgctga tcacgaacct cctcagcgcc aacaaacttt cggagagatg 480
actttggagg attttctcat caaagccgga gttgttcaag aaccatcatc agcatcttcc 540
caacaaaaga aggtggcatc tacccggacc aatggcacaa gtctagaagc aacctacgga 600
atggggcatg gggccggatc ttctcaacag aaaatgttga catctattca aaatagcagt 660
gcaagcttgg atgcaaactt cgggatggga catgtgatgg ggctaggatt cccaggccac 720
caaattgttc ccagcaactt ggcagcacca ggcaatggtt acgccgcagc ataccccatt 780
ttcacgcaga gtaaaactat gatgggggaa tcttctaatg ttgctgaaaa tgggaatggc 840
accaaccgcc tgttagagcc cgtcgtgata cagaacaaga agaggatcat agatggtccg 900
ccggaggtgg tagtcgaaag gagacaacgc cgcatgatca agaatcgaga gtcagcagca 960
agatctcgag caaggaaaca ggcgtacaca gtggaactgg aattagagtt gaaccagctg 1020
aaacaggaga atgcaaagct taagcagctt gtggaggaaa acgagcaaaa gcgaaagcaa 1080
gaggttctga aaagaaaaca atccaaatta ccagcacaaa gaaaggttgt tgataagatg 1140
aggacattga ggaggacagt gagcttggga tggtga 1176
Claims (5)
1.一种培育转基因植物的方法，包括如下步骤：将序列表中序列1所示的氨基酸序列组成的蛋白质的编码基因导入出发植物，得到种子脂肪酸含量提高的转基因植物；所述脂肪酸为18:0脂肪酸、18:1脂肪酸、20:0脂肪酸或20:1脂肪酸；所述植物为棉花或拟南芥。
2.一种提高植物种子脂肪酸含量的方法，包括如下步骤：提高出发植物中序列表中序列1所示的氨基酸序列组成的蛋白质的表达量，得到种子脂肪酸含量提高的植物；所述脂肪酸为18:0脂肪酸、18:1脂肪酸、20:0脂肪酸或20:1脂肪酸；所述植物为棉花或拟南芥。
3.一种培育转基因植物的方法，包括如下步骤：将序列表中序列1所示的氨基酸序列组成的蛋白质的编码基因导入出发植物，得到种子脂肪酸含量降低的转基因植物；所述脂肪酸为16:0脂肪酸、16:1脂肪酸、18:3脂肪酸或22:1脂肪酸；所述植物为棉花或拟南芥。
4.一种降低植物种子脂肪酸含量的方法，包括如下步骤：提高出发植物中序列表中序列1所示的氨基酸序列组成的蛋白质的表达量，得到种子脂肪酸含量降低的植物；所述脂肪酸为16:0脂肪酸、16:1脂肪酸、18:3脂肪酸或22:1脂肪酸；所述植物为棉花或拟南芥。
5.根据权利要求1或3所述的方法，其特征在于：所述编码基因为序列表中序列2所示的DNA分子。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810686038.7A CN108794612B (zh) | 2018-06-28 | 2018-06-28 | 植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810686038.7A CN108794612B (zh) | 2018-06-28 | 2018-06-28 | 植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108794612A CN108794612A (zh) | 2018-11-13 |
| CN108794612B true CN108794612B (zh) | 2021-08-10 |
Family
ID=64071375
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810686038.7A Expired - Fee Related CN108794612B (zh) | 2018-06-28 | 2018-06-28 | 植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108794612B (zh) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102060919A (zh) * | 2009-07-06 | 2011-05-18 | 中国农业科学院生物技术研究所 | 三个棉花abf/areb/abi5/dpbf类转录因子及其编码基因与应用 |
| CN103224551A (zh) * | 2013-05-13 | 2013-07-31 | 中国农业大学 | 与提高种子脂肪酸含量相关的蛋白及其编码基因与应用 |
| WO2015024119A1 (en) * | 2013-08-19 | 2015-02-26 | 8411824 Canada Inc | Plants producing seeds which remove chlorophyl during maturation under stress conditions |
| CN105924512A (zh) * | 2016-07-12 | 2016-09-07 | 中国农业科学院棉花研究所 | 一种与油脂含量相关的GhLPAAT5-like蛋白及其编码基因与应用 |
-
2018
- 2018-06-28 CN CN201810686038.7A patent/CN108794612B/zh not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102060919A (zh) * | 2009-07-06 | 2011-05-18 | 中国农业科学院生物技术研究所 | 三个棉花abf/areb/abi5/dpbf类转录因子及其编码基因与应用 |
| CN103224551A (zh) * | 2013-05-13 | 2013-07-31 | 中国农业大学 | 与提高种子脂肪酸含量相关的蛋白及其编码基因与应用 |
| WO2015024119A1 (en) * | 2013-08-19 | 2015-02-26 | 8411824 Canada Inc | Plants producing seeds which remove chlorophyl during maturation under stress conditions |
| CN105924512A (zh) * | 2016-07-12 | 2016-09-07 | 中国农业科学院棉花研究所 | 一种与油脂含量相关的GhLPAAT5-like蛋白及其编码基因与应用 |
Non-Patent Citations (5)
| Title |
|---|
| "PREDICTED：Gossypium hirsutum ABSCISIC ACID-INSENSITIVE 5-like protein 1 (LOC107889167)，mRNA", NCBI Reference Sequence:XM_016813521.1;genbank;《GenBank》;20160518;第1-2页 * |
| "WRI1-1, ABI5, NF-YA3 and NF-YC2 increase oil biosynthesis in coordination with hormonal signaling during fruit development in oil palm";Wan-Chin Yeap et al.;《The Plant Journal》;20170329;第91卷;第102页右栏第1-2段，图4a * |
| ABA-HYPERSENSITIVE BTB/POZ PROTEIN 1 functions as a negative regulator in ABA-mediated inhibition of germination in Arabidopsis;Hani Kim et al.;《Plant Mol Biol 》;20151214;第90卷;第303-315页 * |
| genbank."PREDICTED：Gossypium hirsutum ABSCISIC ACID-INSENSITIVE 5-like protein 1 (LOC107889167)，mRNA", NCBI Reference Sequence:XM_016813521.1.《GenBank》.2016, * |
| 木本油料植物麻疯树的研究进展;吴平治等;《生命科学》;20140531;第26卷(第5期);第500页左栏第2段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108794612A (zh) | 2018-11-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108948170B (zh) | 一种植物株型生长发育相关蛋白及其编码基因与应用 | |
| CN108642065B (zh) | 一种水稻胚乳粉质相关基因OsSecY2及其编码蛋白质和应用 | |
| CN108440672B (zh) | 一条光呼吸代谢改造支路及其在c3植物中的应用 | |
| CN102757487B (zh) | 植物矮化相关蛋白GA2ox及其编码基因和应用 | |
| CN114560919B (zh) | 一种与植物耐旱相关的转录因子VcMYB108及其编码基因与应用 | |
| CN108794611B (zh) | 植物种子油分相关蛋白GhPDAT1d及其编码基因和应用 | |
| CN108276481B (zh) | 陆地棉GhLEA3基因及其在抗低温胁迫方面的应用 | |
| CN112322648A (zh) | 一种abc转运蛋白基因mrp1s及其制备方法和应用 | |
| CN107475264B (zh) | Dgm1蛋白在提高植物根毛生成能力中的应用 | |
| CN101880674B (zh) | 枣树水通道蛋白基因及在提高植物耐旱性和耐盐性中的应用 | |
| CN111423500B (zh) | SiMYB56蛋白及其编码基因在调控植物耐干旱能力中的应用 | |
| CN110713994A (zh) | 一种植物耐逆性相关蛋白TaMAPK3及其编码基因和应用 | |
| CN110218247B (zh) | PwRBP1和PwNAC1两种蛋白互作协同提高植物耐逆性及其应用 | |
| CN108794612B (zh) | 植物种子脂肪酸相关蛋白GhbZIP67及其编码基因和应用 | |
| CN114539373B (zh) | 与甘薯茎线虫病抗性相关蛋白IbPIF1及其编码基因与应用 | |
| CN109971772B (zh) | 一种耐低温棉花品种的育种方法 | |
| CN112481291B (zh) | GmSAP16蛋白及其编码基因在调控植物耐逆性中的应用 | |
| CN108440658B (zh) | 水稻叶绿体核糖体蛋白编码基因OsWGL2及其用途 | |
| CN106349353B (zh) | 一种调控植物淀粉合成相关蛋白OsFSE及其编码基因与应用 | |
| CN106749582B (zh) | 来源于绿豆的抗旱相关蛋白VrMYB1及其编码基因和应用 | |
| CN111825753A (zh) | 一种植物表观遗传因子及其编码基因在氮素高效利用中的应用 | |
| CN115073572B (zh) | 一种调控植物穗粒数的蛋白AcRR1及其编码基因与应用 | |
| CN111454964A (zh) | 油菜抗寒基因BnTR1及其编码蛋白与应用 | |
| CN114591971B (zh) | 一种葡萄抗旱VvCCD7基因及其氨基酸序列和应用 | |
| CN108409845B (zh) | 蛋白质TaNRT2.5在调控植物对氮肥利用效率中的应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210810 |