CN106520723A - Protein VvMas and encoding gene, and application thereof in improvement of salt tolerance of plants - Google Patents
Protein VvMas and encoding gene, and application thereof in improvement of salt tolerance of plants Download PDFInfo
- Publication number
- CN106520723A CN106520723A CN201611045870.6A CN201611045870A CN106520723A CN 106520723 A CN106520723 A CN 106520723A CN 201611045870 A CN201611045870 A CN 201611045870A CN 106520723 A CN106520723 A CN 106520723A
- Authority
- CN
- China
- Prior art keywords
- plant
- vvmas
- sequence
- gene
- gsp
- 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.)
- Granted
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 75
- 230000015784 hyperosmotic salinity response Effects 0.000 title claims abstract description 28
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 23
- 241000196324 Embryophyta Species 0.000 claims abstract description 80
- 230000009261 transgenic effect Effects 0.000 claims abstract description 38
- 241000894006 Bacteria Species 0.000 claims abstract description 14
- 239000012634 fragment Substances 0.000 claims abstract description 12
- 239000013598 vector Substances 0.000 claims abstract description 10
- 230000014509 gene expression Effects 0.000 claims abstract description 6
- 150000003839 salts Chemical class 0.000 claims description 27
- 108020004414 DNA Proteins 0.000 claims description 22
- 235000018102 proteins Nutrition 0.000 claims description 19
- 238000000034 method Methods 0.000 claims description 15
- 241001233957 eudicotyledons Species 0.000 claims description 6
- 239000013604 expression vector Substances 0.000 claims description 6
- 101100448444 Caenorhabditis elegans gsp-3 gene Proteins 0.000 claims description 5
- 101100448445 Caenorhabditis elegans gsp-4 gene Proteins 0.000 claims description 5
- 101150092780 GSP1 gene Proteins 0.000 claims description 5
- 101150035751 GSP2 gene Proteins 0.000 claims description 5
- 239000012467 final product Substances 0.000 claims description 5
- 241000209510 Liliopsida Species 0.000 claims description 4
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 4
- 125000000539 amino acid group Chemical group 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 239000002773 nucleotide Substances 0.000 claims description 4
- 125000003729 nucleotide group Chemical group 0.000 claims description 4
- 230000008676 import Effects 0.000 claims description 3
- GOJUJUVQIVIZAV-UHFFFAOYSA-N 2-amino-4,6-dichloropyrimidine-5-carbaldehyde Chemical group NC1=NC(Cl)=C(C=O)C(Cl)=N1 GOJUJUVQIVIZAV-UHFFFAOYSA-N 0.000 claims description 2
- 235000001014 amino acid Nutrition 0.000 claims description 2
- 230000008034 disappearance Effects 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 238000009396 hybridization Methods 0.000 claims 1
- 108700026220 vif Genes Proteins 0.000 abstract 1
- 240000007594 Oryza sativa Species 0.000 description 24
- 241000219194 Arabidopsis Species 0.000 description 22
- 235000007164 Oryza sativa Nutrition 0.000 description 14
- 230000012010 growth Effects 0.000 description 13
- 241000219095 Vitis Species 0.000 description 12
- 235000009392 Vitis Nutrition 0.000 description 11
- 239000000463 material Substances 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- 239000002689 soil Substances 0.000 description 10
- 230000009182 swimming Effects 0.000 description 10
- 241000589158 Agrobacterium Species 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 235000009566 rice Nutrition 0.000 description 8
- 239000002585 base Substances 0.000 description 7
- 239000000969 carrier Substances 0.000 description 7
- 239000002299 complementary DNA Substances 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- 238000012216 screening Methods 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 6
- 238000010998 test method Methods 0.000 description 6
- 230000009466 transformation Effects 0.000 description 6
- 206010020649 Hyperkeratosis Diseases 0.000 description 5
- 235000014787 Vitis vinifera Nutrition 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 230000007246 mechanism Effects 0.000 description 5
- 235000016709 nutrition Nutrition 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 240000006365 Vitis vinifera Species 0.000 description 4
- 241000746966 Zizania Species 0.000 description 4
- 239000011543 agarose gel Substances 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 4
- 229940097572 chloromycetin Drugs 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 235000002532 grape seed extract Nutrition 0.000 description 4
- 238000005286 illumination Methods 0.000 description 4
- 238000010369 molecular cloning Methods 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 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 4
- 229960001225 rifampicin Drugs 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 101100437498 Escherichia coli (strain K12) uidA gene Proteins 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 230000004087 circulation Effects 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 238000003754 machining Methods 0.000 description 3
- 210000001161 mammalian embryo Anatomy 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- 230000002018 overexpression Effects 0.000 description 3
- 150000008442 polyphenolic compounds Chemical class 0.000 description 3
- 235000013824 polyphenols Nutrition 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 238000012797 qualification Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 230000002441 reversible effect Effects 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 108090000364 Ligases Proteins 0.000 description 2
- 102000003960 Ligases Human genes 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 108020005089 Plant RNA Proteins 0.000 description 2
- LXVLKXPFIDDHJG-CIUDSAMLSA-N Pro-Glu-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O LXVLKXPFIDDHJG-CIUDSAMLSA-N 0.000 description 2
- 101000702488 Rattus norvegicus High affinity cationic amino acid transporter 1 Proteins 0.000 description 2
- 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 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 235000002636 Zizania aquatica Nutrition 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000007598 dipping method Methods 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003792 electrolyte Substances 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000013467 fragmentation Methods 0.000 description 2
- 238000006062 fragmentation reaction Methods 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- XUWPJKDMEZSVTP-LTYMHZPRSA-N kalafungina Chemical compound O=C1C2=C(O)C=CC=C2C(=O)C2=C1[C@@H](C)O[C@H]1[C@@H]2OC(=O)C1 XUWPJKDMEZSVTP-LTYMHZPRSA-N 0.000 description 2
- 229930027917 kanamycin Natural products 0.000 description 2
- 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 2
- 229960000318 kanamycin Drugs 0.000 description 2
- 229930182823 kanamycin A Natural products 0.000 description 2
- 108010034529 leucyl-lysine Proteins 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 230000000149 penetrating effect Effects 0.000 description 2
- 230000035479 physiological effects, processes and functions Effects 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 238000004153 renaturation Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000011435 rock Substances 0.000 description 2
- 108010026333 seryl-proline Proteins 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 108010015385 valyl-prolyl-proline Proteins 0.000 description 2
- JNTMAZFVYNDPLB-PEDHHIEDSA-N (2S,3S)-2-[[[(2S)-1-[(2S,3S)-2-amino-3-methyl-1-oxopentyl]-2-pyrrolidinyl]-oxomethyl]amino]-3-methylpentanoic acid Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JNTMAZFVYNDPLB-PEDHHIEDSA-N 0.000 description 1
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- DQVAZKGVGKHQDS-UHFFFAOYSA-N 2-[[1-[2-[(2-amino-4-methylpentanoyl)amino]-4-methylpentanoyl]pyrrolidine-2-carbonyl]amino]-4-methylpentanoic acid Chemical compound CC(C)CC(N)C(=O)NC(CC(C)C)C(=O)N1CCCC1C(=O)NC(CC(C)C)C(O)=O DQVAZKGVGKHQDS-UHFFFAOYSA-N 0.000 description 1
- 241001290610 Abildgaardia Species 0.000 description 1
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 1
- YYSWCHMLFJLLBJ-ZLUOBGJFSA-N Ala-Ala-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YYSWCHMLFJLLBJ-ZLUOBGJFSA-N 0.000 description 1
- UCIYCBSJBQGDGM-LPEHRKFASA-N Ala-Arg-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@@H]1C(=O)O)N UCIYCBSJBQGDGM-LPEHRKFASA-N 0.000 description 1
- OQCPATDFWYYDDX-HGNGGELXSA-N Ala-Gln-His Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O OQCPATDFWYYDDX-HGNGGELXSA-N 0.000 description 1
- MDNAVFBZPROEHO-DCAQKATOSA-N Ala-Lys-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O MDNAVFBZPROEHO-DCAQKATOSA-N 0.000 description 1
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 1
- DEWWPUNXRNGMQN-LPEHRKFASA-N Ala-Met-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N1CCC[C@@H]1C(=O)O)N DEWWPUNXRNGMQN-LPEHRKFASA-N 0.000 description 1
- CNQAFFMNJIQYGX-DRZSPHRISA-N Ala-Phe-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 CNQAFFMNJIQYGX-DRZSPHRISA-N 0.000 description 1
- JAQNUEWEJWBVAY-WBAXXEDZSA-N Ala-Phe-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 JAQNUEWEJWBVAY-WBAXXEDZSA-N 0.000 description 1
- WQLDNOCHHRISMS-NAKRPEOUSA-N Ala-Pro-Ile Chemical compound [H]N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WQLDNOCHHRISMS-NAKRPEOUSA-N 0.000 description 1
- VJVQKGYHIZPSNS-FXQIFTODSA-N Ala-Ser-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N VJVQKGYHIZPSNS-FXQIFTODSA-N 0.000 description 1
- 241000219195 Arabidopsis thaliana Species 0.000 description 1
- IGULQRCJLQQPSM-DCAQKATOSA-N Arg-Cys-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(O)=O IGULQRCJLQQPSM-DCAQKATOSA-N 0.000 description 1
- XLWSGICNBZGYTA-CIUDSAMLSA-N Arg-Glu-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O XLWSGICNBZGYTA-CIUDSAMLSA-N 0.000 description 1
- NMRHDSAOIURTNT-RWMBFGLXSA-N Arg-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N NMRHDSAOIURTNT-RWMBFGLXSA-N 0.000 description 1
- SLQQPJBDBVPVQV-JYJNAYRXSA-N Arg-Phe-Val Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O SLQQPJBDBVPVQV-JYJNAYRXSA-N 0.000 description 1
- WKPXXXUSUHAXDE-SRVKXCTJSA-N Arg-Pro-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O WKPXXXUSUHAXDE-SRVKXCTJSA-N 0.000 description 1
- ADPACBMPYWJJCE-FXQIFTODSA-N Arg-Ser-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O ADPACBMPYWJJCE-FXQIFTODSA-N 0.000 description 1
- QCTOLCVIGRLMQS-HRCADAONSA-N Arg-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O QCTOLCVIGRLMQS-HRCADAONSA-N 0.000 description 1
- 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 1
- UTSMXMABBPFVJP-SZMVWBNQSA-N Arg-Val-Trp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N UTSMXMABBPFVJP-SZMVWBNQSA-N 0.000 description 1
- PAXHINASXXXILC-SRVKXCTJSA-N Asn-Asp-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N)O PAXHINASXXXILC-SRVKXCTJSA-N 0.000 description 1
- XBQSLMACWDXWLJ-GHCJXIJMSA-N Asp-Ala-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O XBQSLMACWDXWLJ-GHCJXIJMSA-N 0.000 description 1
- 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 1
- IJHUZMGJRGNXIW-CIUDSAMLSA-N Asp-Glu-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O IJHUZMGJRGNXIW-CIUDSAMLSA-N 0.000 description 1
- SPKCGKRUYKMDHP-GUDRVLHUSA-N Asp-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)O)N SPKCGKRUYKMDHP-GUDRVLHUSA-N 0.000 description 1
- QNMKWNONJGKJJC-NHCYSSNCSA-N Asp-Leu-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O QNMKWNONJGKJJC-NHCYSSNCSA-N 0.000 description 1
- ZQFRDAZBTSFGGW-SRVKXCTJSA-N Asp-Ser-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ZQFRDAZBTSFGGW-SRVKXCTJSA-N 0.000 description 1
- OTKUAVXGMREHRX-CFMVVWHZSA-N Asp-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(O)=O)CC1=CC=C(O)C=C1 OTKUAVXGMREHRX-CFMVVWHZSA-N 0.000 description 1
- ZKQDCIXGCQPQNV-UHFFFAOYSA-N Calcium hypochlorite Chemical compound [Ca+2].Cl[O-].Cl[O-] ZKQDCIXGCQPQNV-UHFFFAOYSA-N 0.000 description 1
- HBHMVBGGHDMPBF-GARJFASQSA-N Cys-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CS)N HBHMVBGGHDMPBF-GARJFASQSA-N 0.000 description 1
- QQOWCDCBFFBRQH-IXOXFDKPSA-N Cys-Phe-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CS)N)O QQOWCDCBFFBRQH-IXOXFDKPSA-N 0.000 description 1
- 108010090461 DFG peptide Proteins 0.000 description 1
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 1
- 206010015866 Extravasation Diseases 0.000 description 1
- CRRFJBGUGNNOCS-PEFMBERDSA-N Gln-Asp-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O CRRFJBGUGNNOCS-PEFMBERDSA-N 0.000 description 1
- KVXVVDFOZNYYKZ-DCAQKATOSA-N Gln-Gln-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O KVXVVDFOZNYYKZ-DCAQKATOSA-N 0.000 description 1
- VUVKKXPCKILIBD-AVGNSLFASA-N Gln-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCC(=O)N)N VUVKKXPCKILIBD-AVGNSLFASA-N 0.000 description 1
- SHAUZYVSXAMYAZ-JYJNAYRXSA-N Gln-Leu-Phe Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CCC(=O)N)N SHAUZYVSXAMYAZ-JYJNAYRXSA-N 0.000 description 1
- FNAJNWPDTIXYJN-CIUDSAMLSA-N Gln-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCC(N)=O FNAJNWPDTIXYJN-CIUDSAMLSA-N 0.000 description 1
- NSEKYCAADBNQFE-XIRDDKMYSA-N Gln-Trp-Arg Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(N)=O)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)=CNC2=C1 NSEKYCAADBNQFE-XIRDDKMYSA-N 0.000 description 1
- BBFCMGBMYIAGRS-AUTRQRHGSA-N Gln-Val-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O BBFCMGBMYIAGRS-AUTRQRHGSA-N 0.000 description 1
- VYOILACOFPPNQH-UMNHJUIQSA-N Gln-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N VYOILACOFPPNQH-UMNHJUIQSA-N 0.000 description 1
- JRCUFCXYZLPSDZ-ACZMJKKPSA-N Glu-Asp-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O JRCUFCXYZLPSDZ-ACZMJKKPSA-N 0.000 description 1
- SWRVAQHFBRZVNX-GUBZILKMSA-N Glu-Lys-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O SWRVAQHFBRZVNX-GUBZILKMSA-N 0.000 description 1
- UJMNFCAHLYKWOZ-DCAQKATOSA-N Glu-Lys-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O UJMNFCAHLYKWOZ-DCAQKATOSA-N 0.000 description 1
- LPHGXOWFAXFCPX-KKUMJFAQSA-N Glu-Pro-Phe Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CCC(=O)O)N)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)O LPHGXOWFAXFCPX-KKUMJFAQSA-N 0.000 description 1
- UZWUBBRJWFTHTD-LAEOZQHASA-N Glu-Val-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCC(O)=O UZWUBBRJWFTHTD-LAEOZQHASA-N 0.000 description 1
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 1
- JPVGHHQGKPQYIL-KBPBESRZSA-N Gly-Phe-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 JPVGHHQGKPQYIL-KBPBESRZSA-N 0.000 description 1
- ZZJVYSAQQMDIRD-UWVGGRQHSA-N Gly-Pro-His Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O ZZJVYSAQQMDIRD-UWVGGRQHSA-N 0.000 description 1
- GAAHQHNCMIAYEX-UWVGGRQHSA-N Gly-Pro-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN GAAHQHNCMIAYEX-UWVGGRQHSA-N 0.000 description 1
- XHVONGZZVUUORG-WEDXCCLWSA-N Gly-Thr-Lys Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CCCCN XHVONGZZVUUORG-WEDXCCLWSA-N 0.000 description 1
- SYOJVRNQCXYEOV-XVKPBYJWSA-N Gly-Val-Glu Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SYOJVRNQCXYEOV-XVKPBYJWSA-N 0.000 description 1
- CTGZVVQVIBSOBB-AVGNSLFASA-N His-His-Glu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(O)=O CTGZVVQVIBSOBB-AVGNSLFASA-N 0.000 description 1
- ZFDKSLBEWYCOCS-BZSNNMDCSA-N His-Phe-Lys Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@@H](N)CC=1NC=NC=1)C1=CC=CC=C1 ZFDKSLBEWYCOCS-BZSNNMDCSA-N 0.000 description 1
- PZAJPILZRFPYJJ-SRVKXCTJSA-N His-Ser-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O PZAJPILZRFPYJJ-SRVKXCTJSA-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
- GQKSJYINYYWPMR-NGZCFLSTSA-N Ile-Gly-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N1CCC[C@@H]1C(=O)O)N GQKSJYINYYWPMR-NGZCFLSTSA-N 0.000 description 1
- CMNMPCTVCWWYHY-MXAVVETBSA-N Ile-His-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CC(C)C)C(=O)O)N CMNMPCTVCWWYHY-MXAVVETBSA-N 0.000 description 1
- PARSHQDZROHERM-NHCYSSNCSA-N Ile-Lys-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)O)N PARSHQDZROHERM-NHCYSSNCSA-N 0.000 description 1
- KTTMFLSBTNBAHL-MXAVVETBSA-N Ile-Phe-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CS)C(=O)O)N KTTMFLSBTNBAHL-MXAVVETBSA-N 0.000 description 1
- OWSWUWDMSNXTNE-GMOBBJLQSA-N Ile-Pro-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(=O)O)C(=O)O)N OWSWUWDMSNXTNE-GMOBBJLQSA-N 0.000 description 1
- SHVFUCSSACPBTF-VGDYDELISA-N Ile-Ser-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N SHVFUCSSACPBTF-VGDYDELISA-N 0.000 description 1
- HXIDVIFHRYRXLZ-NAKRPEOUSA-N Ile-Ser-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)O)N HXIDVIFHRYRXLZ-NAKRPEOUSA-N 0.000 description 1
- KBDIBHQICWDGDL-PPCPHDFISA-N Ile-Thr-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N KBDIBHQICWDGDL-PPCPHDFISA-N 0.000 description 1
- KWHFUMYCSPJCFQ-NGTWOADLSA-N Ile-Thr-Trp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N KWHFUMYCSPJCFQ-NGTWOADLSA-N 0.000 description 1
- 244000017020 Ipomoea batatas Species 0.000 description 1
- 235000002678 Ipomoea batatas Nutrition 0.000 description 1
- UGTHTQWIQKEDEH-BQBZGAKWSA-N L-alanyl-L-prolylglycine zwitterion Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UGTHTQWIQKEDEH-BQBZGAKWSA-N 0.000 description 1
- LHSGPCFBGJHPCY-UHFFFAOYSA-N L-leucine-L-tyrosine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 LHSGPCFBGJHPCY-UHFFFAOYSA-N 0.000 description 1
- CSFVADKICPDRRF-KKUMJFAQSA-N Leu-His-Leu Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C([O-])=O)CC1=CN=CN1 CSFVADKICPDRRF-KKUMJFAQSA-N 0.000 description 1
- KVOFSTUWVSQMDK-KKUMJFAQSA-N Leu-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(C)C)CC1=CN=CN1 KVOFSTUWVSQMDK-KKUMJFAQSA-N 0.000 description 1
- RXGLHDWAZQECBI-SRVKXCTJSA-N Leu-Leu-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O RXGLHDWAZQECBI-SRVKXCTJSA-N 0.000 description 1
- KZZCOWMDDXDKSS-CIUDSAMLSA-N Leu-Ser-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O KZZCOWMDDXDKSS-CIUDSAMLSA-N 0.000 description 1
- JIHDFWWRYHSAQB-GUBZILKMSA-N Leu-Ser-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O JIHDFWWRYHSAQB-GUBZILKMSA-N 0.000 description 1
- VDIARPPNADFEAV-WEDXCCLWSA-N Leu-Thr-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O VDIARPPNADFEAV-WEDXCCLWSA-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
- GGAPIOORBXHMNY-ULQDDVLXSA-N Lys-Arg-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCCN)N)O GGAPIOORBXHMNY-ULQDDVLXSA-N 0.000 description 1
- 108010062166 Lys-Asn-Asp Proteins 0.000 description 1
- KPJJOZUXFOLGMQ-CIUDSAMLSA-N Lys-Asp-Asn Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N KPJJOZUXFOLGMQ-CIUDSAMLSA-N 0.000 description 1
- HIIZIQUUHIXUJY-GUBZILKMSA-N Lys-Asp-Gln Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O HIIZIQUUHIXUJY-GUBZILKMSA-N 0.000 description 1
- YVMQJGWLHRWMDF-MNXVOIDGSA-N Lys-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCCN)N YVMQJGWLHRWMDF-MNXVOIDGSA-N 0.000 description 1
- DTUZCYRNEJDKSR-NHCYSSNCSA-N Lys-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN DTUZCYRNEJDKSR-NHCYSSNCSA-N 0.000 description 1
- LNMKRJJLEFASGA-BZSNNMDCSA-N Lys-Phe-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O LNMKRJJLEFASGA-BZSNNMDCSA-N 0.000 description 1
- MCNGIXXCMJAURZ-VEVYYDQMSA-N Met-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCSC)N)O MCNGIXXCMJAURZ-VEVYYDQMSA-N 0.000 description 1
- AXHNAGAYRGCDLG-UWVGGRQHSA-N Met-Lys-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O AXHNAGAYRGCDLG-UWVGGRQHSA-N 0.000 description 1
- HLZORBMOISUNIV-DCAQKATOSA-N Met-Ser-Leu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(C)C HLZORBMOISUNIV-DCAQKATOSA-N 0.000 description 1
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- DVOCGBNHAUHKHJ-DKIMLUQUSA-N Phe-Ile-Leu 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(C)C)C(O)=O DVOCGBNHAUHKHJ-DKIMLUQUSA-N 0.000 description 1
- KDYPMIZMXDECSU-JYJNAYRXSA-N Phe-Leu-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 KDYPMIZMXDECSU-JYJNAYRXSA-N 0.000 description 1
- YTILBRIUASDGBL-BZSNNMDCSA-N Phe-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 YTILBRIUASDGBL-BZSNNMDCSA-N 0.000 description 1
- MCIXMYKSPQUMJG-SRVKXCTJSA-N Phe-Ser-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MCIXMYKSPQUMJG-SRVKXCTJSA-N 0.000 description 1
- LHALYDBUDCWMDY-CIUDSAMLSA-N Pro-Glu-Ala Chemical compound C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CCCN1)C(O)=O LHALYDBUDCWMDY-CIUDSAMLSA-N 0.000 description 1
- BRJGUPWVFXKBQI-XUXIUFHCSA-N Pro-Leu-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BRJGUPWVFXKBQI-XUXIUFHCSA-N 0.000 description 1
- KWMZPPWYBVZIER-XGEHTFHBSA-N Pro-Ser-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KWMZPPWYBVZIER-XGEHTFHBSA-N 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 108700008625 Reporter Genes Proteins 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- RDFQNDHEHVSONI-ZLUOBGJFSA-N Ser-Asn-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O RDFQNDHEHVSONI-ZLUOBGJFSA-N 0.000 description 1
- 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 1
- UIGMAMGZOJVTDN-WHFBIAKZSA-N Ser-Gly-Ser Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O UIGMAMGZOJVTDN-WHFBIAKZSA-N 0.000 description 1
- MOINZPRHJGTCHZ-MMWGEVLESA-N Ser-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N MOINZPRHJGTCHZ-MMWGEVLESA-N 0.000 description 1
- 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
- HEUVHBXOVZONPU-BJDJZHNGSA-N Ser-Leu-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HEUVHBXOVZONPU-BJDJZHNGSA-N 0.000 description 1
- PPNPDKGQRFSCAC-CIUDSAMLSA-N Ser-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPNPDKGQRFSCAC-CIUDSAMLSA-N 0.000 description 1
- BSXKBOUZDAZXHE-CIUDSAMLSA-N Ser-Pro-Glu Chemical compound [H]N[C@@H](CO)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O BSXKBOUZDAZXHE-CIUDSAMLSA-N 0.000 description 1
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- NJEMRSFGDNECGF-GCJQMDKQSA-N Thr-Ala-Asp Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O NJEMRSFGDNECGF-GCJQMDKQSA-N 0.000 description 1
- TYVAWPFQYFPSBR-BFHQHQDPSA-N Thr-Ala-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)NCC(O)=O TYVAWPFQYFPSBR-BFHQHQDPSA-N 0.000 description 1
- CEXFELBFVHLYDZ-XGEHTFHBSA-N Thr-Arg-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O CEXFELBFVHLYDZ-XGEHTFHBSA-N 0.000 description 1
- RKDFEMGVMMYYNG-WDCWCFNPSA-N Thr-Gln-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O RKDFEMGVMMYYNG-WDCWCFNPSA-N 0.000 description 1
- YOOAQCZYZHGUAZ-KATARQTJSA-N Thr-Leu-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YOOAQCZYZHGUAZ-KATARQTJSA-N 0.000 description 1
- FDQXPJCLVPFKJW-KJEVXHAQSA-N Thr-Met-Tyr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N)O FDQXPJCLVPFKJW-KJEVXHAQSA-N 0.000 description 1
- KERCOYANYUPLHJ-XGEHTFHBSA-N Thr-Pro-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O KERCOYANYUPLHJ-XGEHTFHBSA-N 0.000 description 1
- 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 1
- WCTYCXZYBNKEIV-SXNHZJKMSA-N Trp-Glu-Ile Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O)=CNC2=C1 WCTYCXZYBNKEIV-SXNHZJKMSA-N 0.000 description 1
- OCCYDHCUKXRPSJ-SXNHZJKMSA-N Trp-Ile-Gln Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(O)=O OCCYDHCUKXRPSJ-SXNHZJKMSA-N 0.000 description 1
- ULHASJWZGUEUNN-XIRDDKMYSA-N Trp-Lys-Ser Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O ULHASJWZGUEUNN-XIRDDKMYSA-N 0.000 description 1
- QYSBJAUCUKHSLU-JYJNAYRXSA-N Tyr-Arg-Val Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(O)=O QYSBJAUCUKHSLU-JYJNAYRXSA-N 0.000 description 1
- WPVGRKLNHJJCEN-BZSNNMDCSA-N Tyr-Asp-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 WPVGRKLNHJJCEN-BZSNNMDCSA-N 0.000 description 1
- QAYSODICXVZUIA-WLTAIBSBSA-N Tyr-Gly-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O QAYSODICXVZUIA-WLTAIBSBSA-N 0.000 description 1
- PQPWEALFTLKSEB-DZKIICNBSA-N Tyr-Val-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O PQPWEALFTLKSEB-DZKIICNBSA-N 0.000 description 1
- HHSILIQTHXABKM-YDHLFZDLSA-N Val-Asp-Phe Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](Cc1ccccc1)C(O)=O HHSILIQTHXABKM-YDHLFZDLSA-N 0.000 description 1
- DHINLYMWMXQGMQ-IHRRRGAJSA-N Val-His-His Chemical compound C([C@H](NC(=O)[C@@H](N)C(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 DHINLYMWMXQGMQ-IHRRRGAJSA-N 0.000 description 1
- KISFXYYRKKNLOP-IHRRRGAJSA-N Val-Phe-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)O)N KISFXYYRKKNLOP-IHRRRGAJSA-N 0.000 description 1
- DOFAQXCYFQKSHT-SRVKXCTJSA-N Val-Pro-Pro Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DOFAQXCYFQKSHT-SRVKXCTJSA-N 0.000 description 1
- UJMCYJKPDFQLHX-XGEHTFHBSA-N Val-Ser-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N)O UJMCYJKPDFQLHX-XGEHTFHBSA-N 0.000 description 1
- SDHZOOIGIUEPDY-JYJNAYRXSA-N Val-Ser-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CO)NC(=O)[C@@H](N)C(C)C)C(O)=O)=CNC2=C1 SDHZOOIGIUEPDY-JYJNAYRXSA-N 0.000 description 1
- OWFGFHQMSBTKLX-UFYCRDLUSA-N Val-Tyr-Tyr Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N OWFGFHQMSBTKLX-UFYCRDLUSA-N 0.000 description 1
- JSOXWWFKRJKTMT-WOPDTQHZSA-N Val-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N JSOXWWFKRJKTMT-WOPDTQHZSA-N 0.000 description 1
- LLJLBRRXKZTTRD-GUBZILKMSA-N Val-Val-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N LLJLBRRXKZTTRD-GUBZILKMSA-N 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 108010050025 alpha-glutamyltryptophan Proteins 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000007844 bleaching agent Substances 0.000 description 1
- 230000009172 bursting Effects 0.000 description 1
- 229940027138 cambia Drugs 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000002390 cell membrane structure Anatomy 0.000 description 1
- UCKZMPLVLCKKMO-LHLIQPBNSA-N cephamycin Chemical compound S1CC(C)=C(C(O)=O)N2C(=O)[C@@H](C)[C@]21OC UCKZMPLVLCKKMO-LHLIQPBNSA-N 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- KXZOIWWTXOCYKR-UHFFFAOYSA-M diclofenac potassium Chemical compound [K+].[O-]C(=O)CC1=CC=CC=C1NC1=C(Cl)C=CC=C1Cl KXZOIWWTXOCYKR-UHFFFAOYSA-M 0.000 description 1
- 108010054812 diprotin A Proteins 0.000 description 1
- 108010054813 diprotin B Proteins 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 230000036251 extravasation Effects 0.000 description 1
- 238000010230 functional analysis Methods 0.000 description 1
- 238000003208 gene overexpression Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 1
- 239000007952 growth promoter Substances 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 108010027338 isoleucylcysteine Proteins 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010091871 leucylmethionine Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 108010012058 leucyltyrosine Proteins 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 235000019362 perlite Nutrition 0.000 description 1
- 239000010451 perlite Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 108010004914 prolylarginine Proteins 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000012879 subculture medium Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 238000000844 transformation Methods 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 108010000998 wheylin-2 peptide Proteins 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Plant Pathology (AREA)
- Medicinal Chemistry (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a protein VvMas, an encoding gene, a recombinant vector, expression box, transgenic cell line or recombinant bacterium containing the encoding gene, and a primer pair for amplifying the full length or any fragment of the encoding gene, and also provides an application of the protein VvMas, the encoding gene, and the recombinant vector, expression box, transgenic cell line or recombinant bacterium containing the encoding gene in improvement of the salt tolerance of plants. The protein and the encoding gene thereof have important application values in cultivation of salt-tolerance plant varieties in order to improve output of crops, and have wide application space and wide market prospect in the agriculture field.
Description
Technical field
The present invention relates to a kind of plant salt endurance associated protein VvMas and its encoding gene and application, particularly derive from
Salt tolerance associated protein VvMas of Fructus Vitis viniferae and its encoding gene and application.
Background technology
There is large-area saliferous soil in the world.According to statistics, the whole world has 800,000,000 hm2Salt-soda soil, in irrigated land
There is the secondary salinization soil for accounting for cultivated area 33% in area, and the bursting of salt of soil has a strong impact on the development of modern agriculture.With regard to me
For state, there is nearly 1/10th secondary salinization soil in the arable land of 1,800,000,000 mu of the whole nation, in addition also 20,000,000 hm2Salt alkaline wasteland
Ground.In general, salinity can affect the growth of crop, but the salinity in salt-soda soil mostly in 0.6%- in 0.2%-0.5%
10%.The presence of large-area salinization land has had a strong impact on grain-production, becomes the principal element for limiting agricultural production.With
The sharp increase of world population and the decline year by year of arable area, grain-production receives safely serious threat, for ploughing per capita
The relatively small China of ground area is even more increasingly serious problem.By the further investigation to plant salt tolerance mechanism, salt tolerant is cultivated
New Crop Varieties be using saline alkali land resource is most economical, one of effective measures.
The Mechanism of Salt-tolerant of plant is considerably complicated, and it is related to growth promoter, morphosiss, physiological feature and Metabolism regulation
Etc. all many-sides.When plant is subject to salt stress, the aspect such as the form of plant, Physiology and biochemistry can all occur a series of change,
Its existence could be maintained.Salt damage suppresses plant tissue, the growth of organ and differentiation, affects plant cell membrane structure, makes cell membrane
Permeability increases, and reduces photosynthetic rate, causes a large amount of electrolyte and non-electrolyte extravasation, film fat component to change, memebrane protein
Component and activity are affected, and then affect the physiological metabolism of plant.Therefore plant is in long-term evolution and adaptation process, by
Step defines a series of mechanism for resisting extraneous poor environment change.
Plant under condition of salt stress can go to prevent or mitigate the harm of salt using certain strategy, in long-term evolution
During, plant has developed a series of Mechanisms of Salt Resistance.With developing rapidly for molecular biology, plant salt tolerance Physiology and biochemistry
Mechanism is increasingly clear and definite so that clone is possibly realized with plant salt tolerance related gene.Strengthen the physiological research of plant salt tolerance, verify plant
Vital movement rule of the thing under adverse circumstance in addition artificial adjustment, cultivate the improved seeds with opposing poor environment character, with
The yield and quality of crop is improved, agricultural year stable yields is significant for obtaining.
The content of the invention
Technical problem：In order to solve the defect of prior art, the invention provides a kind of plant salt endurance associated protein
VvMas and its encoding gene, additionally provide the application of above-mentioned albumen.
Technical scheme：The present invention provide with plant salt endurance associated protein, its entitled VvMas (α/β hydrolytic enzyme), come
It is following (a) or (b) to come from Fructus Vitis viniferae (Vitis vinifera)：
A () is by sequence SEQ ID NO in sequence table：The protein of the aminoacid sequence composition shown in 2；
B () is by sequence SEQ ID NO in sequence table：2 amino acid residue sequence is through one or several amino acid residues
Replacement and/or disappearance and/or addition and related to plant salt endurance by SEQ ID NO：Protein derived from 2.
Present invention also offers the gene of encoding proteins VvMas.
The gene, with C N metabolism associated protein, is arbitrary described gene in following (1)-(3)：
(1) its nucleotides sequence is classified as SEQ ID NO：DNA molecular shown in 1；
(2) DNA sequence for being limited with (1) under strict conditions is hybridized and the DNA of coded plant salt tolerance associated protein divides
Son；
(3) DNA sequence limited with (1) or (2) at least with 70%, at least with 75%, at least with 80%, at least
With 85%, at least with 90%, at least with 95%, at least with 96%, at least with 97%, at least have 98% or
At least there is the DNA molecular of 99% homology and coded plant salt tolerance associated protein.
Above-mentioned stringent condition can be, with 6 × SSC, the solution of 0.5%SDS, to hybridize at 65 DEG C, then with 2 × SSC,
0.1%SDS and 1 × SSC, 0.1%SDS respectively wash film once.
Sequence SEQ ID NO：1 by 1224 base compositions, sequence SEQ ID NO in polynucleotide：Egg shown in 2
In vain.
Present invention also offers containing it is described with the expression cassette of the encoding gene of plant salt endurance associated protein, it is recombinant expressed
Carrier, transgenic cell line or recombinant bacterium.
The recombinant expression carrier is to insert to obtain final product said gene in expression vector.
Specifically, the recombinant vector is to insert the encoding gene to obtain between the multiple clone site of carrier pCBGUS
's.
Wherein, the carrier pCBGUS is that method by comprising the steps is obtained：
(1) by pCAMBIA1301 carriers through Hind III and EcoR I double digestions, reclaim carrier large fragment；
(2) 121 carriers of pBI are reclaimed into the fragment comprising gusA genes through Hind III and EcoR I double digestions；
(3) fragment comprising gusA genes by the carrier large fragment reclaimed in step (1) with recovery in step (2) connects
Connect, obtain final product carrier pCBGUS.
The pCAMBIA1301 carriers are purchased from CAMBIA companies；121 carriers of the pBI are purchased from Clontech companies.
Present invention also offers the amplification encoding gene total length or its arbitrary fragment with plant salt endurance associated protein
Primer pair.
Specifically, the primer pair sequence is as follows：
GSP-1：5’-CTGAGACGAGTTGGGGTGGAA-3’
GSP-2：5’-GTTTTCCCAGTCACGAC-3’
GSP-3：5’-GACTTGGCCTCCAGGTTGACCTTGA-3’
GSP-4：5’-GGCCACGCGTCGACTAGTACGGGGGGGGGG-3’
GSP-5：5’-ATGAAAGGCGTCTTTTCGGCGCCAG-3’
GSP-6：5’-TTACACAAGACATCTACTTTTCCAA-3’
Present invention also offers albumen VvMas, its encoding gene or its recombinant vector, expression cassette, transgenic cell line, weight
Application of the group bacterium in plant salt endurance is improved；The plant is dicotyledon or monocotyledon.
Present invention also offers a kind of method for cultivating transgenic plant, comprises the following steps：By egg described in claim 1
White encoding gene imports purpose plant, obtains final product.
Specifically, the encoding gene of albumen described in claim 1 is led by the recombinant vector described in claim 4 or 5
Enter purpose plant；The plant is specially dicotyledon or monocotyledon.
The purpose plant is dicotyledon or monocotyledon；The preferred arabidopsiss of the dicotyledon or Oryza sativa L..
Beneficial effect：The albumen of the VvMas coded by said gene that the present invention is provided can improve the salt tolerance of plant, with weight
The using value wanted, the research for improving Fructus Vitis viniferae salt tolerance provide important foundation.
The albumen and its encoding gene of the present invention has important using value to cultivating Salt tolerant plants kind, so as to carry
High crop yield is significant, will have wide application space and market prospect in agriculture field.
Description of the drawings
Fig. 1 Fructus Vitis viniferae VvMas gene plants expression vector sketches of the present invention.
The PCR testing result figures of Fig. 2 VvMas transgenic Arabidopsis plants of the present invention.
The PCR testing result figures of Fig. 3 VvMas transgenic rice plants of the present invention.
The potted plant qualification figure of salt tolerance of Fig. 4 VvMas transgenic Arabidopsis plants of the present invention.
The salt tolerance Detached-leaf test figure of Fig. 5 VvMas transgenic rice plants of the present invention.
The salt tolerance water planting qualification figure of Fig. 6 VvMas transgenic rice plants of the present invention.
The potted plant qualification figure of salt tolerance of Fig. 7 VvMas transgenic rice plants of the present invention.
Specific embodiment
In following embodiments, test material used and its source include：
Fructus Vitis viniferae (Vitis vinifera) kind PN40024, by Huaiyingong College life sciences and food engineering institute river
Su Sheng plant production and the laboratory preservation of machining Practice Education Center.
The seed of arabidopsiss (Arabidopsis thaliana) is through 2.5% (v/v) CaClO2It is planted in after sterilization black
Soil：Vermiculitum：Perlite (1:1:1) in mixed-matrix, 22 DEG C, 16h illumination cultivation (16h illumination, 8h are dark, cold light source) growth
2 weeks.
No. 11 are spent in Oryza sativa L. (Oryza sativa) kind, by Huaiyingong College life sciences and food engineering institute Jiangsu
Plant production is saved with the laboratory preservation of machining Practice Education Center.
Escherichia coli (Escherichia coli) DH5 α are by Huaiyingong College life sciences and food engineering institute Jiangsu
Plant production is saved with the laboratory preservation of machining Practice Education Center.Cloning vehicle PMD-18-Simple T, all kinds of restriction enzymes
Enzyme, Taq polymerase, ligase, dNTP, 10 × PCR buffer and DNA marker are purchased from the limited public affairs in precious biological engineering Dalian
Department.All of chemical reagent is all bought from sigma chemical company of the U.S. and Shanghai traditional Chinese medicines chemical reagents corporation.
In the present invention conventional molecular biology manipulations referring specifically to《Molecular cloning》【Molecular Cloning.2nd
ed.Cold Spring Harbor Laboratory Press,1989】。
In following embodiments, conventional genetic manipulation is carried out with reference to molecular cloning document【Sambook J,Frets EF,
Mannsdes T et al.In:Molecular Cloning.2nd ed.Cold Spring Harbor Laboratory
Press,1989】。
The acquisition of the related albumen of 1 Fructus Vitis viniferae salt tolerance of embodiment and its encoding gene
1. experiment material
Grape variety PN40024 aseptic seedling expansion leaf blade is removed, liquid nitrogen flash freezer, -80 DEG C of preservations.
2. blade Total RNAs extraction and purification
Take PN40024 aseptic seedling and launch leaf blade about 2.0g, grind to form powdery in liquid nitrogen, add 10mL centrifuge tubes, use
It is total that Applygen plant RNA extraction test kits (Applygen Technologies Inc, Beijing) extract sweet potato root tuber
RNA, test kit include：Plant RNA Reagent, plant tissue cracking, separation RNA, removal vegetable polysaccharidess and polyphenol；
Extraction Reagent, Organic extraction remove isolating protein, DNA, polysaccharide and polyphenol；Plant RNA Aid, remove plant many
Sugared polyphenol and secondary metabolite.Using QIAGEN Oligotex Mini mRNA Kit (QIAGEN, GmbH, Germany) from
Purified mRNA in total serum IgE.Finally, 1 μ L are taken and its integrity is detected in 1.2% agarose gel electrophoresiies, separately take 2 μ L and be diluted to 500 μ
L, with its quality of UV spectrophotometer measuring (OD260) and purity (OD260/OD280), the PN40024 aseptic seedling leaves of extraction
Piece total serum IgE, the detection of Jing non denatured glue agarose gel electrophoresiies, 28S and 18S bands are clear, and the two brightness ratio is 1.5～2
1, show that total serum IgE is not degraded, obtained by purification, mRNA meets requirement of experiment, can be used for Fructus Vitis viniferae VvMas albumen cDNA total lengths gram
It is grand.
The full-length clone of 3.VvMas albumen cDNA
The full-length clone of VvMas albumen cDNA is carried out with the VvMas EST fragments design primer that this laboratory is obtained.
(1)3’-RACE
With PN40024cDNA as template, enter performing PCR with VvMas EST forward primers GSP-1 and reverse primer GSP-2 anti-
Should.Primer sequence is as follows：
GSP-1：5’-CTGAGACGAGTTGGGGTGGAA-3’
GSP-2：5’-GTTTTCCCAGTCACGAC-3’
3 ' the RACE fragments that PCR is obtained, connect after recovery pMD19-T carriers (purchased from the logical Trade Co., Ltd. of Beijing six directions,
Catalog number is D102A) TA clones are carried out, draw so that BcaBESTTM Sequencing Primers/M13Primers are general
Thing is sequenced.
(2)5’-RACE
With PN40024cDNA as template, enter performing PCR with VvMas EST forward primers GSP-3 and reverse primer GSP-4 anti-
Should.Primer sequence is as follows：
GSP-3：5’-GACTTGGCCTCCAGGTTGACCTTGA-3’
GSP-4：5’-GGCCACGCGTCGACTAGTACGGGGGGGGGG-3’
5 ' the RACE fragments that PCR is obtained, connect after recovery pMD19-T carriers (purchased from the logical Trade Co., Ltd. of Beijing six directions,
Catalog number is D102A) TA clones are carried out, draw so that BcaBESTTM Sequencing Primers/M13Primers are general
Thing is sequenced.
(3) PCR expands the coding region of VvMas albumen cDNA
Splice the Fructus Vitis viniferae VvMas Protein cDNA Sequences of candidate using 7.0 softwares of DNAMAN.Forward primer is designed further
GSP-5 and reverse primer GSP-6 enter the coding region that performing PCR expands VvMas albumen cDNA.Primer sequence is as follows：
GSP-5：5’-ATGAAAGGCGTCTTTTCGGCGCCAG-3’
GSP-6：5’-TTACACAAGACATCTACTTTTCCAA-3’
With PN40024 blade total serum IgE Jing Oligo (dT) reverse transcriptions as template, with the FastPfu enzymes of high-fidelity, enter performing PCR
Amplification, PCR conditions are 95 DEG C of 1min, subsequent 95 DEG C of 20s, 53 DEG C of 20s and 72 DEG C of 1min, carry out 40 circulations, and last 72 DEG C are prolonged
Stretch 10min.Agarose gel electrophoresiies detect pcr amplification product, obtain the amplified fragments of 1224bp length.
The result of summary step, obtains purpose cDNA sequence, sequence SEQ in its nucleotide sequence such as sequence table
ID NO：Shown in 1.Sequence SEQ ID NO in sequence table：1 by 1224 base compositions, from 5 ' the 1st the-the 1224 bit bases in end
For its open reading frame, coding is with sequence SEQ ID NO in sequence table：The protein of the amino acid residue sequence shown in 2.Sequence
Sequence SEQ ID NO in list：2 are made up of 407 amino acid residues.By the unnamed gene be VvMas, the albumen for being encoded
It is named as VvMas.
The structure of 2 VvMas gene overexpression carriers of embodiment
Sequencing identification in embodiment 1 is correctly contained sequence table SEQ ID NO：The DNA fragmentation of nucleotide shown in 1 is used
BamH I and Sac I carries out double digestion, reclaims DNA fragmentation with 1% agarose gel, by T4DNA ligase by reclaim
VvMas genetic fragments are connected with double 35S promoter pYPx245 plasmids are contained, and enzyme action identification and sequence analysis are determined and contained
There is the recombiant plasmid AH128 of Fructus Vitis viniferae VvMas genes.The expression vector also includes gusA reporter genes and band intron kanamycin
Resistant maker gene, carrier are as shown in Figure 1.
Embodiment 3VvMas gene transformation arabidopsiss
The plant expression vector pCAMBIA1301-VvMas of the Fructus Vitis viniferae VvMas genes that embodiment 2 is built is with dipping in Hua Fazhuan
Change arabidopsiss, concrete grammar is as follows：
1. the preparation of Agrobacterium
(1) pCAMBIA1301-VvMas is converted into agrobacterium tumefaciens lba4404 bacterial strain (Biovector with electric shocking method
Co., LTD), the recombinational agrobacterium containing pCAMBIA1301-VvMas is obtained, and coats the flat board containing kalamycin resistance
Screening transformant.
(2) picking Agrobacterium list bacterium is inoculated in 5mL LB fluid mediums (50 μ g/mL of rifampicin, 100 μ g/mL of chloromycetin)
In, 28 DEG C, 250rpm culture 20h.
(3) take 1mL bacterium solutions to transfer into 20-30mL LB fluid mediums (50 μ g/mL of rifampicin, 100 μ g/mL of chloromycetin)
In, 28 DEG C, 250rpm culture about 12h survey 600 ≈ 1.5 of OD.
(4) 8000rpm, 4 DEG C, 10min is collected by centrifugation thalline, be resuspended in Agrobacterium-mediated Transformation penetrating fluid (5% sucrose,
0.05%Silwet L-77) and it is diluted to 600 ≈ 0.8 of OD.
2. arabidopsiss dip in colored method conversion
(1) infect colored a kind of sedge immersion of arabidopsiss above-mentioned in liquid, take out after being gently agitated for about 10s, be totally converted after finishing,
Arabidopsiss are covered with freshness protection package, to keep moist environment, horizontal positioned, 22 DEG C of lucifuge cultures to remove freshness protection package after 24h and uprightly train
Support.
(2) after first transfonning four days, can once be converted again, be repeated twice, total cotransformation three times so can be to flower
The alabastrum of the different times developed in sequence is converted, and improves transformation efficiency.
(3) growth about after two months, collects seed, and 4 DEG C of refrigerator storages are stand-by.
Arabidopsiss through dipping in colored method conversion grow about after two months, knot of normally blooming.
3 VvMas genes of embodiment turn Oryza sativa L.
The plant expression vector pCAMBIA1301-VvMas rice transformations of the Fructus Vitis viniferae VvMas genes that embodiment 2 is built,
Concrete grammar is as follows：
1. the preparation of Agrobacterium
(1) by pCAMBIA1301-VvMas with electric shocking method conversion Agrobacterium tumefaciems EHA105 bacterial strains (Biovector Co.,
LTD), the recombinational agrobacterium containing pCAMBIA1301-VvMas is obtained, and coats the plate screening containing kalamycin resistance
Transformant.
(2) picking Agrobacterium list bacterium is inoculated in 5mL LB fluid mediums (50 μ g/mL of rifampicin, 100 μ g/mL of chloromycetin)
In, 28 DEG C, 250rpm culture 20h.
(3) take 1mL bacterium solutions to transfer into 20-30mL LB fluid mediums (50 μ g/mL of rifampicin, 100 μ g/mL of chloromycetin)
In, 28 DEG C, 250rpm culture about 12h survey 600 ≈ 1.5 of OD.
(4) 8000rpm, 4 DEG C, 10min is collected by centrifugation thalline, be resuspended in Agrobacterium-mediated Transformation penetrating fluid (5% sucrose,
0.05%Silwet L-77) and it is diluted to 600 ≈ 0.8 of OD.
2. the acquisition of Mature Embryos of Rice wound healing
(1) No. 11 seeds will be spent to remove clever shell in ripe rice varieties, with 70% alcohol disinfecting 1-2min；
(2) then with 20% sodium hypochlorite immersion 30-40min, with sterile distilled water rinse 4 times, seed is transferred to
Surface moisture is blotted on sterilized filter paper, is then seeded on NB inducing cultures；
(3) after light culture 7-10d, when scultellum expands, when endosperm softens, remove embryo and bud, the embryo callus subculture for peeling is turned
Move on on NB subculture mediums, about 3w subcultures once, can serve as receptor after subculture 2-3 time and be converted.
3. agriculture stalk bacterium mediated transformation Rice Callus
(1) choose good embryo callus be put in it is above-mentioned infect in liquid, soak 30min；
(2) calluss are taken out, unnecessary bacterium solution is sucked with aseptic filter paper, be subsequently placed in NB and co-culture culture in culture medium
There is (about 2-3d) to just there is bacterium colony；
(3) with sterilized water oscillation cleaning 3-4 time, till supernatant is cleaned completely, with 500mg/L cephamycin solution
Oscillation cleaning 40min；
(4) calluss are taken out, 0.4m/s air-dries 4h in being put into the only sterile petri dish with filter paper, proceeds to NB screening and culturing
Base screening two-wheeled (often taking turns 3-4w)；
(5) kanamycin-resistant callus tissue is carried out into pre- differentiation 2-3w, is then transferred into illumination cultivation 2-3w in division culture medium；
(6) strong seedling culture base culture 30d or so is proceeded to when plumelet length is to about 1cm；
(7) throw off the culture of sealed membrane seedling exercising one week or so, be then transplanted in soil.
4 VvMas gene transgenic Arabidopsis plants PCR of embodiment is detected
1. the screening of transgenic arabidopsis seed
(1) 25-30mg seeds are claimed to be put into 1.5mL centrifuge tubes；
(2) 75% ethanol disinfection 1min of 1mL (not stopping to rock vibration), 8000rpm centrifugation 5s, removes supernatant；
(3) bleaching powder (2.5%) sterilization 15min (not stopping to rock vibration, sufficiently sterilised) after adding 1mL to filter,
8000rpm is centrifuged 5s, removes supernatant；
(4) aseptic water washing 3-4 time；
(5) seed is uniformly sowed on 1/2MS flat boards (50 μ g/mL of hygromycin), the sealing of Parafilm films, 4 DEG C of ice
Case placement two days, 22 DEG C, 16h illumination cultivation 10 days.
(6) resistant plant is transplanted in basin and is cultivated, after Seedling is slightly larger, carried out GUS Activity determinations, select positive plant (T1)
Cultivate to blossoming and bearing fruit, collect T1Tied T on plant2Seed, further screening obtain T3Seed.
2. transgenic Arabidopsis plants PCR detections
(1) test method
T is extracted with CTAB methods3The genomic DNA of arabidopsiss transfer-gen plant and WT lines.Carried out with conventional method
PCR detects that the VvMas gene primers for being used are：primer 1：5 '-ACAGCGTCTCCGACCTGATGCA-3 ' and
primer2：5’-AGTCAATGACCGCTGTTATGCG-3’.10 × PCR is added in 0.2mL Eppendorf centrifuge tubes
21 μ L of μ L, 4dNTP (10mol/L) of buffer, primer (10 μm of ol/L) are 1 μ L, template DNA (50ng/uL) 2 μ L, Taq
0.25 μ L of archaeal dna polymerase, plus ddH2O to 20 μ L of cumulative volume.Response procedures be 94 DEG C of denaturations 5min, 94 DEG C of degeneration 30s, 55 DEG C
Renaturation 30s, 72 DEG C of extension 2min, totally 35 circulations.
(2) result of the test
Electrophoresis detection amplification is shown in that (in Fig. 2, swimming lane M is Maker to Fig. 2；Swimming lane W：Water；Swimming lane P：Positive control (restructuring
Plasmid pCAMBIA1301-VvMas)；Swimming lane Col-0：Wild-type Arabidopsis plants；Swimming lane #1-#5：For conversion
The arabidopsiss transfer-gen plant of pCAMBIA1301-VvMas).It can be seen that the arabidopsiss of conversion pCAMBIA1301-VvMas
Intend the target stripe that transfer-gen plant and positive control amplify 591bp, show that VvMas genes have been incorporated into the base of arabidopsiss
In because of group, and prove that these regeneration plants are transfer-gen plant；Wild-type Arabidopsis plants do not amplify the target bar of 591bp
Band.Transfer-gen plant is analyzed for follow-up function.
5 VvMas gene transgenic rice plants PCR of embodiment is detected
(1) test method
T is extracted with CTAB methods2The genomic DNA of Transgenic Rice Plants and WT lines.Enter performing PCR with conventional method
Detect, the VvMas gene primers for being used are：primer 1：5 '-ACAGCGTCTCCGACCTGATGCA-3 ' and primer2：
5’-AGTCAATGACCGCTGTTATGCG-3’.Add in 0.2mL Eppendorf centrifuge tubes 10 × PCR buffer, 2 μ L,
1 μ L of 4dNTP (10mol/L), primer (10 μm of ol/L) are 1 μ L, 2 μ L, Taq archaeal dna polymerase 0.25 of template DNA (50ng/uL)
μ L, plus ddH2O to 20 μ L of cumulative volume.Response procedures be 94 DEG C of denaturations 5min, 94 DEG C of degeneration 30s, 55 DEG C of renaturation 30s, 72 DEG C
Extend 2min, totally 35 circulations.
(2) result of the test
Electrophoresis detection amplification is shown in that (in Fig. 3, swimming lane M is Maker to Fig. 3；Swimming lane W：Water；Swimming lane P：Positive control (restructuring
Plasmid pCAMBIA1301-VvMas)；Swimming lane WT：Wild rice plant；Swimming lane OE1-OE11：For converting pCAMBIA1301-
The Transgenic Rice Plants of VvMas).It can be seen that the Oryza sativa L. of conversion pCAMBIA1301-VvMas intends transfer-gen plant and sun
Property control amplify the target stripe of 591bp, show that VvMas genes have been incorporated in the genome of Oryza sativa L., and prove these
Regeneration plant is transfer-gen plant；Wild rice plant does not amplify the target stripe of 591bp.Transfer-gen plant is follow-up
Functional analyses.
6 VvMas gene transgenic Arabidopsis plant Salt-Tolerance Identifications of embodiment
(1) test method
After transgenic arabidopsis and wild type seeds are cultivated 2w in 1/2MS culture medium, plantlet of transplant is trained in basin
After foster 2w, salt stress process is carried out.With containing 300mM NaCl 1/2 suddenly each 2d of Glan nutritional solution irrigate 1 time, every time
200mL, processes 4w, observes plant strain growth situation and takes a picture, counts survival rate.
(2) result of the test
As a result show, by the potted plant identification of salt tolerance, as a result see Fig. 4, after salt treatment 4w, the growth conditions of transfer-gen plant
WT lines are significantly better than, the survival rate of transfer-gen plant is significantly higher than wildness plant.Show that overexpression VvMas genes show
Write the salt tolerance for improving transgenic Arabidopsis plants.
7 VvMas gene transgenic rice plant Salt-Tolerance Identifications of embodiment
1. transgenic rice plant growth potential is compared
(1) test method
The seed disinfection of transgenic paddy rice material and wild type material, seed are sowed on MS solid plates, seed is sent out
The seed of germination state consistency after bud 2-3d, is chosen, the different middle finger pipe cultures in MS and MS+NaCl (200mM) are sowed respectively
On base, after Seedling growth 7-10d, the Seedling growing way of different disposal starts difference occur, carries out photograph and growth potential statistics, including Seedling
Long and fresh weight data.
(2) result of the test
As a result show, under salt stress treatment conditions, as a result see Fig. 5, transgenic line and wild type WT materials because
The presence of condition of salt stress, plant diminish；But transgenic line is compared with wild type WT, and growth conditions are relatively preferable, growth
Gesture data statisticss show that the Seedling length of transgenic line and fresh weight are superior to wild type WT materials.
2. transgenic rice plant salt tolerance water planting identification
(1) test method
By the T of homozygosis2Transgenic paddy rice and wild rice seed are seeded in MS culture medium, grow about 3-4d.Choose
The seedling that growing way is consistent is selected, is transplanted in 96 hole PCR plates (its bottom has been cut up), one time of nutrition liquid is changed every 1d, kept away
Exempt from the long bacterium of Seedling root, after normal growth 4w；Stress treatment 2w is carried out in being transferred into 200mM NaCl Hoagland solutions, is observed
Its phenotype, is taken a picture and is investigated its survival rate.
(2) result of the test
As a result show, after salt stress treatment conditions 2w, as a result see Fig. 6, transgenic line is compared with wild type material, poor
It is different it is obvious that the growth conditions of transgenic line are significantly better than wild type material, the survival rate of transfer-gen plant is significantly higher than open country
Natural disposition plant.Show that overexpression VvMas genes significantly improve the salt tolerance of transgenic rice plant.
3. the potted plant identification of transgenic rice plant salt tolerance
(1) test method
In order to further verify the salt tolerance of transgenic paddy rice material, by the T of homozygosis2Transgenic paddy rice and wild rice
The surface of the seed is sterilized, and uses pure water accelerating germination, is seeded in MS culture medium, grows about 3-4d.The consistent seedling of growing way is selected, is planted
It is implanted in Nutrition Soil：Vermiculitum=1:In 2 Nutrition Soil, note watering daily, salt is proceeded by until plant grows to 4 true leaves
Stress treatment.With containing 200mM NaCl 1/2 each 2d irrigation of Glan nutritional solution 1 time suddenly, each 200mL processes 2w, observation
Its phenotype, is taken a picture and is investigated its survival rate.
(2) result of the test
After Fig. 7 results are displayed in salt stress treatment conditions 2w, salt tolerant potted plant experiment shows transgenic line with wild section bar
Material is compared, it is obvious that the growth conditions of transgenic line are significantly better than wild type material, the survival rate of transfer-gen plant shows difference
Write higher than wildness plant.Show that overexpression VvMas genes significantly improve the salt tolerance of transgenic rice plant.
<110>Huaiyingong College
<120>Albumen VvMas, encoding gene and its application in plant salt endurance is improved
<130> 20161118002
<160> 8
<170> PatentIn version 3.3
<210> 1
<211> 1224
<212> DNA
<213> Vitis vinifera
<220>
<221> misc_feature
<222> (1)..(1224)
<400> 1
atgaaaggcg tcttttcggc gccaggagat tacatccact tcaagtctca ggtccctctt 60
cacaagatcc ccattggcac aaagcagtgg cggtattatg attttggtcc aaaagtggtc 120
cctccactta tttgtcttcc tgggacagca ggaactgcag atgtctatta caaacagata 180
atgtcattgt ccataaaggg ttaccgggtg atctctgttg atattcctcg cgtatggaac 240
catcatgagt ggattcaagc atttgaaaag tttttggatg ctattgatgt gcaccatata 300
catctttatg gcacatccct tggaggcttc ctggcacaac tttttgctca gcatcgtcca 360
agacgggtta ggtcgttgat tctctcaaat tcatttttgg agacacgcag tttctcatct 420
gcaatgccat gggcccctat tgtcagttgg accccttctt ttttgctgaa gagatatgtc 480
ttaacgggaa ttccagatgg cccccatgaa ccatttattg cagattcagt agactttgtt 540
gtttcccagg ttgaaacact ctcaagagag gacttggcct ccaggttgac cttgactgtt 600
gatgctgctt ccattggacc tcttcttctc tcagattcat tcatcactct aatggataca 660
aatgactact gttcaattcc tcaacaactc aaagatcagc tgagtgaaag gtaccctgga 720
gcaaggcgag catacttaaa aactggaggt gatttcccat ttctttcacg ttcagatgaa 780
gtaaacctac atcttcagtt acacctgaga cgagttgggg tggaagcccg tccagatttg 840
gtcaagggta tctcaaagga tggtagtggt gggagttcta gtgaaaagaa tgatgaaaga 900
gaagattcag atgttcctcc taaggacaat gggggaagtc ctgaaagccc ttccactgaa 960
acccaattgc cagaggctcc agaaagttcg ggttctcata gcttagatga ccagctcctc 1020
agcaatgcaa aggtttgctt caccagtcct gaacatgtga ggcttcccat atctcatgca 1080
ttttttgaga aacagcaaga cattgtatcc acaaggtttg tccaaccggc ttgggagatt 1140
tttattcttt gtctgcttcc tctctatgtg gaaacaatgt acattacttg gattttttgt 1200
tggaaaagta gatgtcttgt gtaa 1224
<210> 2
<211> 407
<212> PRT
<213> Vitis vinifera
<220>
<221> MISC_FEATURE
<222> (1)..(407)
<400> 2
Met Lys Gly Val Phe Ser Ala Pro Gly Asp Tyr Ile His Phe Lys Ser
1 5 10 15
Gln Val Pro Leu His Lys Ile Pro Ile Gly Thr Lys Gln Trp Arg Tyr
20 25 30
Tyr Asp Phe Gly Pro Lys Val Val Pro Pro Leu Ile Cys Leu Pro Gly
35 40 45
Thr Ala Gly Thr Ala Asp Val Tyr Tyr Lys Gln Ile Met Ser Leu Ser
50 55 60
Ile Lys Gly Tyr Arg Val Ile Ser Val Asp Ile Pro Arg Val Trp Asn
65 70 75 80
His His Glu Trp Ile Gln Ala Phe Glu Lys Phe Leu Asp Ala Ile Asp
85 90 95
Val His His Ile His Leu Tyr Gly Thr Ser Leu Gly Gly Phe Leu Ala
100 105 110
Gln Leu Phe Ala Gln His Arg Pro Arg Arg Val Arg Ser Leu Ile Leu
115 120 125
Ser Asn Ser Phe Leu Glu Thr Arg Ser Phe Ser Ser Ala Met Pro Trp
130 135 140
Ala Pro Ile Val Ser Trp Thr Pro Ser Phe Leu Leu Lys Arg Tyr Val
145 150 155 160
Leu Thr Gly Ile Pro Asp Gly Pro His Glu Pro Phe Ile Ala Asp Ser
165 170 175
Val Asp Phe Val Val Ser Gln Val Glu Thr Leu Ser Arg Glu Asp Leu
180 185 190
Ala Ser Arg Leu Thr Leu Thr Val Asp Ala Ala Ser Ile Gly Pro Leu
195 200 205
Leu Leu Ser Asp Ser Phe Ile Thr Leu Met Asp Thr Asn Asp Tyr Cys
210 215 220
Ser Ile Pro Gln Gln Leu Lys Asp Gln Leu Ser Glu Arg Tyr Pro Gly
225 230 235 240
Ala Arg Arg Ala Tyr Leu Lys Thr Gly Gly Asp Phe Pro Phe Leu Ser
245 250 255
Arg Ser Asp Glu Val Asn Leu His Leu Gln Leu His Leu Arg Arg Val
260 265 270
Gly Val Glu Ala Arg Pro Asp Leu Val Lys Gly Ile Ser Lys Asp Gly
275 280 285
Ser Gly Gly Ser Ser Ser Glu Lys Asn Asp Glu Arg Glu Asp Ser Asp
290 295 300
Val Pro Pro Lys Asp Asn Gly Gly Ser Pro Glu Ser Pro Ser Thr Glu
305 310 315 320
Thr Gln Leu Pro Glu Ala Pro Glu Ser Ser Gly Ser His Ser Leu Asp
325 330 335
Asp Gln Leu Leu Ser Asn Ala Lys Val Cys Phe Thr Ser Pro Glu His
340 345 350
Val Arg Leu Pro Ile Ser His Ala Phe Phe Glu Lys Gln Gln Asp Ile
355 360 365
Val Ser Thr Arg Phe Val Gln Pro Ala Trp Glu Ile Phe Ile Leu Cys
370 375 380
Leu Leu Pro Leu Tyr Val Glu Thr Met Tyr Ile Thr Trp Ile Phe Cys
385 390 395 400
Trp Lys Ser Arg Cys Leu Val
405
<210> 3
<211> 21
<212> DNA
<213> Artificial
<220>
<223> GSP-1
<220>
<221> misc_feature
<222> (1)..(21)
<400> 3
ctgagacgag ttggggtgga a 21
<210> 4
<211> 17
<212> DNA
<213> Artificial
<220>
<223> GSP-2
<220>
<221> misc_feature
<222> (1)..(17)
<400> 4
gttttcccag tcacgac 17
<210> 5
<211> 25
<212> DNA
<213> Artificial
<220>
<223> GSP-3
<220>
<221> misc_feature
<222> (1)..(25)
<400> 5
gacttggcct ccaggttgac cttga 25
<210> 6
<211> 30
<212> DNA
<213> Artificial
<220>
<223> GSP-4
<220>
<221> misc_feature
<222> (1)..(30)
<400> 6
ggccacgcgt cgactagtac gggggggggg 30
<210> 7
<211> 25
<212> DNA
<213> Artificial
<220>
<223> GSP-5
<220>
<221> misc_feature
<222> (1)..(25)
<400> 7
atgaaaggcg tcttttcggc gccag 25
<210> 8
<211> 25
<212> DNA
<213> Artificial
<220>
<223> GSP-6
<220>
<221> misc_feature
<222> (1)..(25)
<400> 8
ttacacaaga catctacttt tccaa 25
Claims (10)
1. a kind of albumen, is following (a) or (b)：
A () is by sequence SEQ ID NO in sequence table：The protein of the aminoacid sequence composition shown in 2；
B () is by sequence SEQ ID NO in sequence table：2 amino acid residue sequence is taken through one or several amino acid residues
Generation and/or disappearance and/or add and related to plant salt endurance by SEQ ID NO：Protein derived from 2.
2. the gene of albumen described in claim 1 is encoded.
3. gene as claimed in claim 2, it is characterised in that：The gene is DNA point of any one in following (1)-(3)
Son；
(1) its nucleotides sequence is classified as SEQ ID NO：DNA molecular shown in 1；
(2) the DNA sequence hybridization for limiting with (1) under strict conditions and the DNA molecular of coded plant salt tolerance associated protein；
(3) DNA sequence limited with (1) or (2) is at least had 70%, at least has 75%, at least have 80%, at least have
85%th, at least with 90%, at least with 95%, at least with 96%, at least with 97%, at least have 98% or at least
With 99% homology and coded plant salt tolerance associated protein DNA molecular.
4. the recombinant vector, expression cassette, transgenic cell line or recombinant bacterium containing gene described in Claims 2 or 3.
5. recombinant vector as claimed in claim 4, it is characterised in that：The recombinant vector is by base described in Claims 2 or 3
Obtain final product in expression vector because inserting.
6. the primer pair of full length gene or its any fragment described in Claims 2 or 3 is expanded.
7. primer pair as claimed in claim 6, its sequence are as follows：
GSP-1：5’-CTGAGACGAGTTGGGGTGGAA-3’；
GSP-2：5’-GTTTTCCCAGTCACGAC-3’；
GSP-3：5’-GACTTGGCCTCCAGGTTGACCTTGA-3’；
GSP-4：5’-GGCCACGCGTCGACTAGTACGGGGGGGGGG-3’；
GSP-5：5’-ATGAAAGGCGTCTTTTCGGCGCCAG-3’；
GSP-6：5’-TTACACAAGACATCTACTTTTCCAA-3’.
8. encoding gene described in albumen, Claims 2 or 3 described in claim 1 or recombinant vector, expression described in claim 4
The application of box, transgenic cell line or recombinant bacterium in plant salt endurance is improved；The plant is dicotyledon or unifacial leaf
Plant.
9. it is a kind of cultivate transgenic plant method, it is characterised in that：Comprise the following steps：By albumen described in claim 1
Encoding gene imports purpose plant, obtains final product.
10. method according to claim 9, it is characterised in that：The encoding gene of albumen described in claim 1 is by power
Profit requires that the recombinant vector described in 4 or 5 imports purpose plant；The plant is specially dicotyledon or monocotyledon.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611045870.6A CN106520723B (en) | 2016-11-22 | 2016-11-22 | Protein VvMas, coding gene and application of protein VvMas in improving salt tolerance of plants |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611045870.6A CN106520723B (en) | 2016-11-22 | 2016-11-22 | Protein VvMas, coding gene and application of protein VvMas in improving salt tolerance of plants |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106520723A true CN106520723A (en) | 2017-03-22 |
| CN106520723B CN106520723B (en) | 2020-01-21 |
Family
ID=58356686
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611045870.6A Active CN106520723B (en) | 2016-11-22 | 2016-11-22 | Protein VvMas, coding gene and application of protein VvMas in improving salt tolerance of plants |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106520723B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107663232A (en) * | 2017-10-27 | 2018-02-06 | 淮阴工学院 | Plant anti-adversity associated protein OsIAA18 and its encoding gene and application |
| CN108047320A (en) * | 2018-02-13 | 2018-05-18 | 淮阴工学院 | The application of Protein G mMYB12 and encoding gene in isoflavones accumulation and salt tolerance is improved |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103204915A (en) * | 2013-04-11 | 2013-07-17 | 中国农业大学 | Salt resistance related protein IbEST of sweet potato and coding gene and application thereof |
-
2016
- 2016-11-22 CN CN201611045870.6A patent/CN106520723B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103204915A (en) * | 2013-04-11 | 2013-07-17 | 中国农业大学 | Salt resistance related protein IbEST of sweet potato and coding gene and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| DEGAO LIU ET AL.: "A Novel a/b-Hydrolase Gene IbMas Enhances Salt Tolerance in Transgenic Sweetpotato", 《PLOS ONE》 * |
| 无: "XM_002267775.4", 《GENBANK》 * |
| 无: "XP_002267811.1", 《GENBANK》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107663232A (en) * | 2017-10-27 | 2018-02-06 | 淮阴工学院 | Plant anti-adversity associated protein OsIAA18 and its encoding gene and application |
| CN107663232B (en) * | 2017-10-27 | 2019-09-24 | 淮阴工学院 | Plant anti-adversity associated protein OsIAA18 and its encoding gene and application |
| CN108047320A (en) * | 2018-02-13 | 2018-05-18 | 淮阴工学院 | The application of Protein G mMYB12 and encoding gene in isoflavones accumulation and salt tolerance is improved |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106520723B (en) | 2020-01-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106868021B (en) | Gene OsNAC1 for controlling rice seed size and application thereof | |
| CN110628808B (en) | Arabidopsis AtTCP5 gene and application thereof in regulating plant height | |
| CN107383179B (en) | A kind of and plant stress tolerance correlative protein GsSLAH3 and its encoding gene and application | |
| CN110643618A (en) | Jatropha curcas MYB transcription factor JcMYB16 gene and application thereof in improving drought resistance of plants | |
| CN110128514B (en) | Rice cold tolerance associated protein CTB4b in booting stage, coding gene and application | |
| CN110004156A (en) | GhCML20 gene relevant to resistance to verticillium wilt and its application | |
| CN114480431A (en) | Application of corn ZmBES1/BZR1-10 gene in improving drought tolerance and yield of plants | |
| CN102154321B (en) | Method for breeding stress-resistance transgenic rice | |
| CN103451192A (en) | Populus deltoidesx populus nigra PdMYB2 gene and application thereof | |
| CN113322261B (en) | Application of soybean ABC transporter gene GmALS3 in breeding of low-phosphorus-resistant and aluminum-toxicity-resistant plants | |
| CN102226188A (en) | Application of paddy rice transcription factor OsAP21 gene | |
| CN116891862B (en) | Zoysia japonica salt tolerance gene ZmLA1, protein and application thereof | |
| CN101698854A (en) | Application of transcription thellungiella halophila CBF1 gene in improving drought resistance and salt tolerance of corn and wheat | |
| CN106892973A (en) | Plant adversity resistance related protein GhMYB4 and encoding gene and application | |
| CN106520723A (en) | Protein VvMas and encoding gene, and application thereof in improvement of salt tolerance of plants | |
| CN103602688B (en) | Helianthus tuberosus L. Na<+>/H<+> reverse transport protein genes HtNHX1 and HtNHX2 and use thereof | |
| CN107663232B (en) | Plant anti-adversity associated protein OsIAA18 and its encoding gene and application | |
| CN115851660A (en) | Application of protein OsPIS and coding gene in improving plant stress resistance | |
| CN111454966B (en) | Cymbidium CgWRKY4 gene and application thereof | |
| CN107630021A (en) | The albumen and application of asparagus resistant gene of salt AoSOS2 and its coding | |
| CN107417780A (en) | The application of UBC32 albumen and its encoding gene in drought resistance in plants is regulated and controled | |
| CN107142266B (en) | ZmRCI2-8 gene and application thereof in promoting plant germination and lateral root growth under abiotic stress condition | |
| CN106336453B (en) | A kind of verticillium wilt resistance of cotton by same GAP-associated protein GAP GaRPL18 and its encoding gene and application | |
| CN111171124A (en) | Plant stress resistance related protein VvIAA18, coding gene and application | |
| CN104341491B (en) | Drought tolerant associated protein for plant OsERF62 and its encoding gene and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | 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 | ||
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20170322 Assignee: Shuyang Yize Landscape Engineering Co.,Ltd. Assignor: HUAIYIN INSTITUTE OF TECHNOLOGY Contract record no.: X2020980008884 Denomination of invention: Protein vvmas, coding genes and their application in improving salt tolerance of plants Granted publication date: 20200121 License type: Common License Record date: 20201208 |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20231123 Address after: 250000 Hengda Yueting, No. 58, Gongye North Road, Licheng District, Jinan City, Shandong Province Patentee after: Dongdai (Jinan) Intelligent Technology Co.,Ltd. Address before: 223005 Huaian 1 Jiangsu economic and Technological Development Zone Patentee before: HUAIYIN INSTITUTE OF TECHNOLOGY |