CN1056881C - 编码杀虫剂蛋白的基因、用该基因转化的禾本科植物及其制备方 - Google Patents
编码杀虫剂蛋白的基因、用该基因转化的禾本科植物及其制备方 Download PDFInfo
- Publication number
- CN1056881C CN1056881C CN92115178A CN92115178A CN1056881C CN 1056881 C CN1056881 C CN 1056881C CN 92115178 A CN92115178 A CN 92115178A CN 92115178 A CN92115178 A CN 92115178A CN 1056881 C CN1056881 C CN 1056881C
- Authority
- CN
- China
- Prior art keywords
- gene
- codon
- usage
- ser
- dna
- 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
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 146
- 230000000749 insecticidal effect Effects 0.000 title claims abstract description 12
- 238000004519 manufacturing process Methods 0.000 title abstract description 3
- 108700005078 Synthetic Genes Proteins 0.000 claims abstract description 42
- 108700010070 Codon Usage Proteins 0.000 claims abstract description 34
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims description 51
- 108020004705 Codon Proteins 0.000 claims description 47
- 150000001413 amino acids Chemical class 0.000 claims description 40
- 244000025254 Cannabis sativa Species 0.000 claims description 13
- 101100449439 Drosophila melanogaster grass gene Proteins 0.000 claims description 13
- 239000000203 mixture Substances 0.000 claims description 13
- 230000008901 benefit Effects 0.000 claims description 7
- 101150057622 grass gene Proteins 0.000 claims description 3
- 241000196324 Embryophyta Species 0.000 abstract description 53
- 241000238631 Hexapoda Species 0.000 abstract description 16
- 210000001938 protoplast Anatomy 0.000 abstract description 13
- 239000013604 expression vector Substances 0.000 abstract description 12
- 239000007788 liquid Substances 0.000 abstract description 8
- 230000001131 transforming effect Effects 0.000 abstract description 8
- 210000004748 cultured cell Anatomy 0.000 abstract description 5
- 240000007594 Oryza sativa Species 0.000 abstract 1
- 230000001172 regenerating effect Effects 0.000 abstract 1
- 239000013598 vector Substances 0.000 abstract 1
- 108020004414 DNA Proteins 0.000 description 62
- 239000012634 fragment Substances 0.000 description 54
- 238000003752 polymerase chain reaction Methods 0.000 description 49
- 241000209094 Oryza Species 0.000 description 46
- 108700012359 toxins Proteins 0.000 description 37
- 229940024606 amino acid Drugs 0.000 description 36
- 235000001014 amino acid Nutrition 0.000 description 36
- 239000003053 toxin Substances 0.000 description 34
- 231100000765 toxin Toxicity 0.000 description 34
- 239000013612 plasmid Substances 0.000 description 32
- 235000018102 proteins Nutrition 0.000 description 23
- 230000014509 gene expression Effects 0.000 description 22
- 102000053602 DNA Human genes 0.000 description 21
- 239000001963 growth medium Substances 0.000 description 20
- 238000002360 preparation method Methods 0.000 description 19
- 108091034117 Oligonucleotide Proteins 0.000 description 16
- 239000002609 medium Substances 0.000 description 16
- 210000004027 cell Anatomy 0.000 description 15
- 108091008146 restriction endonucleases Proteins 0.000 description 14
- 239000000243 solution Substances 0.000 description 14
- 238000006243 chemical reaction Methods 0.000 description 13
- 235000007164 Oryza sativa Nutrition 0.000 description 11
- 235000009566 rice Nutrition 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- 206010020649 Hyperkeratosis Diseases 0.000 description 10
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 10
- 239000012530 fluid Substances 0.000 description 10
- 230000008569 process Effects 0.000 description 9
- 229920000936 Agarose Polymers 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- 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 7
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 7
- 238000010367 cloning Methods 0.000 description 7
- 229940088598 enzyme Drugs 0.000 description 7
- 238000012216 screening Methods 0.000 description 7
- 230000004083 survival effect Effects 0.000 description 7
- 230000009466 transformation Effects 0.000 description 7
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 6
- 101100364969 Dictyostelium discoideum scai gene Proteins 0.000 description 6
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 6
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 6
- 101100364971 Mus musculus Scai gene Proteins 0.000 description 6
- 241000208125 Nicotiana Species 0.000 description 6
- 102000043276 Oncogene Human genes 0.000 description 6
- 108700020796 Oncogene Proteins 0.000 description 6
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 6
- 239000006035 Tryptophane Substances 0.000 description 6
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 6
- 241001233957 eudicotyledons Species 0.000 description 6
- 229930182817 methionine Natural products 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- 239000000725 suspension Substances 0.000 description 6
- 230000007704 transition Effects 0.000 description 6
- 238000013519 translation Methods 0.000 description 6
- 230000014616 translation Effects 0.000 description 6
- 229960004799 tryptophan Drugs 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 5
- 101100016593 Encephalitozoon cuniculi (strain GB-M1) HD-1 gene Proteins 0.000 description 5
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 5
- 241000209510 Liliopsida Species 0.000 description 5
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 5
- 108020005038 Terminator Codon Proteins 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 238000013016 damping Methods 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 230000029087 digestion Effects 0.000 description 5
- 238000001962 electrophoresis Methods 0.000 description 5
- 239000002158 endotoxin Substances 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 230000035772 mutation Effects 0.000 description 5
- 239000001103 potassium chloride Substances 0.000 description 5
- 235000011164 potassium chloride Nutrition 0.000 description 5
- 238000012545 processing Methods 0.000 description 5
- 239000011541 reaction mixture Substances 0.000 description 5
- 108010077805 Bacterial Proteins Proteins 0.000 description 4
- 241000098289 Cnaphalocrocis medinalis Species 0.000 description 4
- 241000008892 Cnaphalocrocis patnalis Species 0.000 description 4
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 4
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 4
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 4
- 238000002105 Southern blotting Methods 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 108010081551 glycylphenylalanine Proteins 0.000 description 4
- 229940027941 immunoglobulin g Drugs 0.000 description 4
- 230000008929 regeneration Effects 0.000 description 4
- 238000011069 regeneration method Methods 0.000 description 4
- 230000008521 reorganization Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 241000701489 Cauliflower mosaic virus Species 0.000 description 3
- 229920000742 Cotton Polymers 0.000 description 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 3
- LMKYZBGVKHTLTN-NKWVEPMBSA-N D-nopaline Chemical compound NC(=N)NCCC[C@@H](C(O)=O)N[C@@H](C(O)=O)CCC(O)=O LMKYZBGVKHTLTN-NKWVEPMBSA-N 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 3
- 241000219146 Gossypium Species 0.000 description 3
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 3
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 3
- 229930195725 Mannitol Natural products 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 3
- JAWGSPUJAXYXJA-IHRRRGAJSA-N Ser-Phe-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CO)N)CC1=CC=CC=C1 JAWGSPUJAXYXJA-IHRRRGAJSA-N 0.000 description 3
- 240000003768 Solanum lycopersicum Species 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000001976 enzyme digestion Methods 0.000 description 3
- 229960002989 glutamic acid Drugs 0.000 description 3
- 238000005286 illumination Methods 0.000 description 3
- 239000000594 mannitol Substances 0.000 description 3
- 235000010355 mannitol Nutrition 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 229920003023 plastic Polymers 0.000 description 3
- 239000004033 plastic Substances 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 2
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 description 2
- 239000004475 Arginine Substances 0.000 description 2
- HOBNTSHITVVNBN-ZPFDUUQYSA-N Asp-Ile-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)O)NC(=O)[C@H](CC(=O)O)N HOBNTSHITVVNBN-ZPFDUUQYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 108010053835 Catalase Proteins 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 2
- 102000012410 DNA Ligases Human genes 0.000 description 2
- 108010061982 DNA Ligases Proteins 0.000 description 2
- 238000007399 DNA isolation Methods 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- KXTAGESXNQEZKB-DZKIICNBSA-N Glu-Phe-Val Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C(C)C)C(O)=O)CC1=CC=CC=C1 KXTAGESXNQEZKB-DZKIICNBSA-N 0.000 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- FFALDIDGPLUDKV-ZDLURKLDSA-N Gly-Thr-Ser Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O FFALDIDGPLUDKV-ZDLURKLDSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- PJLLMGWWINYQPB-PEFMBERDSA-N Ile-Asn-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N PJLLMGWWINYQPB-PEFMBERDSA-N 0.000 description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 2
- 241000880493 Leptailurus serval Species 0.000 description 2
- ILJREDZFPHTUIE-GUBZILKMSA-N Leu-Asp-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ILJREDZFPHTUIE-GUBZILKMSA-N 0.000 description 2
- ZDJQVSIPFLMNOX-RHYQMDGZSA-N Leu-Thr-Arg 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)CCCN=C(N)N ZDJQVSIPFLMNOX-RHYQMDGZSA-N 0.000 description 2
- 235000007688 Lycopersicon esculentum 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
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- VGQVAVQWKJLIRM-FXQIFTODSA-N Ser-Ser-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O VGQVAVQWKJLIRM-FXQIFTODSA-N 0.000 description 2
- YEDSOSIKVUMIJE-DCAQKATOSA-N Ser-Val-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O YEDSOSIKVUMIJE-DCAQKATOSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- LXWZOMSOUAMOIA-JIOCBJNQSA-N Thr-Asn-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N)O LXWZOMSOUAMOIA-JIOCBJNQSA-N 0.000 description 2
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 2
- 239000004473 Threonine Substances 0.000 description 2
- -1 Xie Ansuan Chemical compound 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 229920002494 Zein Polymers 0.000 description 2
- 108010055615 Zein Proteins 0.000 description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 2
- 108010013835 arginine glutamate Proteins 0.000 description 2
- 229960005261 aspartic acid Drugs 0.000 description 2
- 235000003704 aspartic acid Nutrition 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 229940106157 cellulase Drugs 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000013599 cloning vector Substances 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000002425 crystallisation Methods 0.000 description 2
- 230000008025 crystallization Effects 0.000 description 2
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 2
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 2
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 description 2
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 2
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 2
- 230000008034 disappearance Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000006062 fragmentation reaction Methods 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 108010010147 glycylglutamine Proteins 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 108010002685 hygromycin-B kinase Proteins 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 229960000310 isoleucine Drugs 0.000 description 2
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 2
- 238000009533 lab test Methods 0.000 description 2
- 229910001629 magnesium chloride Inorganic materials 0.000 description 2
- 210000001161 mammalian embryo Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 2
- 229960005190 phenylalanine Drugs 0.000 description 2
- 239000013600 plasmid vector Substances 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000010455 vermiculite Substances 0.000 description 2
- 229910052902 vermiculite Inorganic materials 0.000 description 2
- 235000019354 vermiculite Nutrition 0.000 description 2
- 239000005019 zein Substances 0.000 description 2
- 229940093612 zein Drugs 0.000 description 2
- MEIRRNXMZYDVDW-MQQKCMAXSA-N (2E,4E)-2,4-hexadien-1-ol Chemical compound C\C=C\C=C\CO MEIRRNXMZYDVDW-MQQKCMAXSA-N 0.000 description 1
- OJHZNMVJJKMFGX-RNWHKREASA-N (4r,4ar,7ar,12bs)-9-methoxy-3-methyl-1,2,4,4a,5,6,7a,13-octahydro-4,12-methanobenzofuro[3,2-e]isoquinoline-7-one;2,3-dihydroxybutanedioic acid Chemical compound OC(=O)C(O)C(O)C(O)=O.O=C([C@@H]1O2)CC[C@H]3[C@]4([H])N(C)CC[C@]13C1=C2C(OC)=CC=C1C4 OJHZNMVJJKMFGX-RNWHKREASA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- ZEMPKEQAKRGZGQ-AAKVHIHISA-N 2,3-bis[[(z)-12-hydroxyoctadec-9-enoyl]oxy]propyl (z)-12-hydroxyoctadec-9-enoate Chemical compound CCCCCCC(O)C\C=C/CCCCCCCC(=O)OCC(OC(=O)CCCCCCC\C=C/CC(O)CCCCCC)COC(=O)CCCCCCC\C=C/CC(O)CCCCCC ZEMPKEQAKRGZGQ-AAKVHIHISA-N 0.000 description 1
- OVSKIKFHRZPJSS-DOMIDYPGSA-N 2-(2,4-dichlorophenoxy)acetic acid Chemical compound OC(=O)[14CH2]OC1=CC=C(Cl)C=C1Cl OVSKIKFHRZPJSS-DOMIDYPGSA-N 0.000 description 1
- AAQGRPOPTAUUBM-ZLUOBGJFSA-N Ala-Ala-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O AAQGRPOPTAUUBM-ZLUOBGJFSA-N 0.000 description 1
- JBVSSSZFNTXJDX-YTLHQDLWSA-N Ala-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N JBVSSSZFNTXJDX-YTLHQDLWSA-N 0.000 description 1
- YSMPVONNIWLJML-FXQIFTODSA-N Ala-Asp-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(O)=O YSMPVONNIWLJML-FXQIFTODSA-N 0.000 description 1
- ROLXPVQSRCPVGK-XDTLVQLUSA-N Ala-Glu-Tyr Chemical compound N[C@@H](C)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O ROLXPVQSRCPVGK-XDTLVQLUSA-N 0.000 description 1
- YHKANGMVQWRMAP-DCAQKATOSA-N Ala-Leu-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N YHKANGMVQWRMAP-DCAQKATOSA-N 0.000 description 1
- LDLSENBXQNDTPB-DCAQKATOSA-N Ala-Lys-Arg Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N LDLSENBXQNDTPB-DCAQKATOSA-N 0.000 description 1
- ADSGHMXEAZJJNF-DCAQKATOSA-N Ala-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C)N ADSGHMXEAZJJNF-DCAQKATOSA-N 0.000 description 1
- BHTBAVZSZCQZPT-GUBZILKMSA-N Ala-Pro-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C)N BHTBAVZSZCQZPT-GUBZILKMSA-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
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical class N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- GXCSUJQOECMKPV-CIUDSAMLSA-N Arg-Ala-Gln Chemical compound C[C@H](NC(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O GXCSUJQOECMKPV-CIUDSAMLSA-N 0.000 description 1
- NABSCJGZKWSNHX-RCWTZXSCSA-N Arg-Arg-Thr Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H]([C@H](O)C)C(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N NABSCJGZKWSNHX-RCWTZXSCSA-N 0.000 description 1
- CPSHGRGUPZBMOK-CIUDSAMLSA-N Arg-Asn-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O CPSHGRGUPZBMOK-CIUDSAMLSA-N 0.000 description 1
- VXXHDZKEQNGXNU-QXEWZRGKSA-N Arg-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N VXXHDZKEQNGXNU-QXEWZRGKSA-N 0.000 description 1
- JUWQNWXEGDYCIE-YUMQZZPRSA-N Arg-Gln-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O JUWQNWXEGDYCIE-YUMQZZPRSA-N 0.000 description 1
- NKBQZKVMKJJDLX-SRVKXCTJSA-N Arg-Glu-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NKBQZKVMKJJDLX-SRVKXCTJSA-N 0.000 description 1
- AUFHLLPVPSMEOG-YUMQZZPRSA-N Arg-Gly-Glu Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O AUFHLLPVPSMEOG-YUMQZZPRSA-N 0.000 description 1
- RFXXUWGNVRJTNQ-QXEWZRGKSA-N Arg-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCCN=C(N)N)N RFXXUWGNVRJTNQ-QXEWZRGKSA-N 0.000 description 1
- NVUIWHJLPSZZQC-CYDGBPFRSA-N Arg-Ile-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O NVUIWHJLPSZZQC-CYDGBPFRSA-N 0.000 description 1
- AGVNTAUPLWIQEN-ZPFDUUQYSA-N Arg-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N AGVNTAUPLWIQEN-ZPFDUUQYSA-N 0.000 description 1
- LLUGJARLJCGLAR-CYDGBPFRSA-N Arg-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](CCCN=C(N)N)N LLUGJARLJCGLAR-CYDGBPFRSA-N 0.000 description 1
- COXMUHNBYCVVRG-DCAQKATOSA-N Arg-Leu-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O COXMUHNBYCVVRG-DCAQKATOSA-N 0.000 description 1
- YFHATWYGAAXQCF-JYJNAYRXSA-N Arg-Pro-Phe Chemical compound NC(N)=NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 YFHATWYGAAXQCF-JYJNAYRXSA-N 0.000 description 1
- DNLQVHBBMPZUGJ-BQBZGAKWSA-N Arg-Ser-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O DNLQVHBBMPZUGJ-BQBZGAKWSA-N 0.000 description 1
- ICRHGPYYXMWHIE-LPEHRKFASA-N Arg-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O ICRHGPYYXMWHIE-LPEHRKFASA-N 0.000 description 1
- VJIQPOJMISSUPO-BVSLBCMMSA-N Arg-Trp-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O VJIQPOJMISSUPO-BVSLBCMMSA-N 0.000 description 1
- BWMMKQPATDUYKB-IHRRRGAJSA-N Arg-Tyr-Asn Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(N)=O)C(O)=O)CC1=CC=C(O)C=C1 BWMMKQPATDUYKB-IHRRRGAJSA-N 0.000 description 1
- YNDLOUMBVDVALC-ZLUOBGJFSA-N Asn-Ala-Ala Chemical compound C[C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](CC(=O)N)N YNDLOUMBVDVALC-ZLUOBGJFSA-N 0.000 description 1
- CIBWFJFMOBIFTE-CIUDSAMLSA-N Asn-Arg-Gln Chemical compound C(C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)CN=C(N)N CIBWFJFMOBIFTE-CIUDSAMLSA-N 0.000 description 1
- ACRYGQFHAQHDSF-ZLUOBGJFSA-N Asn-Asn-Asn Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O ACRYGQFHAQHDSF-ZLUOBGJFSA-N 0.000 description 1
- BGINHSZTXRJIPP-FXQIFTODSA-N Asn-Asp-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N BGINHSZTXRJIPP-FXQIFTODSA-N 0.000 description 1
- UPALZCBCKAMGIY-PEFMBERDSA-N Asn-Gln-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UPALZCBCKAMGIY-PEFMBERDSA-N 0.000 description 1
- NVWJMQNYLYWVNQ-BYULHYEWSA-N Asn-Ile-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O NVWJMQNYLYWVNQ-BYULHYEWSA-N 0.000 description 1
- IBLAOXSULLECQZ-IUKAMOBKSA-N Asn-Ile-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC(N)=O IBLAOXSULLECQZ-IUKAMOBKSA-N 0.000 description 1
- JTXVXGXTRXMOFJ-FXQIFTODSA-N Asn-Pro-Asn Chemical compound NC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O JTXVXGXTRXMOFJ-FXQIFTODSA-N 0.000 description 1
- XTMZYFMTYJNABC-ZLUOBGJFSA-N Asn-Ser-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)N XTMZYFMTYJNABC-ZLUOBGJFSA-N 0.000 description 1
- NPZJLGMWMDNQDD-GHCJXIJMSA-N Asn-Ser-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NPZJLGMWMDNQDD-GHCJXIJMSA-N 0.000 description 1
- FMNBYVSGRCXWEK-FOHZUACHSA-N Asn-Thr-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O FMNBYVSGRCXWEK-FOHZUACHSA-N 0.000 description 1
- SKQTXVZTCGSRJS-SRVKXCTJSA-N Asn-Tyr-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O SKQTXVZTCGSRJS-SRVKXCTJSA-N 0.000 description 1
- XEGZSHSPQNDNRH-JRQIVUDYSA-N Asn-Tyr-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XEGZSHSPQNDNRH-JRQIVUDYSA-N 0.000 description 1
- SLHOOKXYTYAJGQ-XVYDVKMFSA-N Asp-Ala-His Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 SLHOOKXYTYAJGQ-XVYDVKMFSA-N 0.000 description 1
- NECWUSYTYSIFNC-DLOVCJGASA-N Asp-Ala-Phe Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 NECWUSYTYSIFNC-DLOVCJGASA-N 0.000 description 1
- HMQDRBKQMLRCCG-GMOBBJLQSA-N Asp-Arg-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HMQDRBKQMLRCCG-GMOBBJLQSA-N 0.000 description 1
- YNCHFVRXEQFPBY-BQBZGAKWSA-N Asp-Gly-Arg Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N YNCHFVRXEQFPBY-BQBZGAKWSA-N 0.000 description 1
- LDGUZSIPGSPBJP-XVYDVKMFSA-N Asp-His-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CC(=O)O)N LDGUZSIPGSPBJP-XVYDVKMFSA-N 0.000 description 1
- DWOGMPWRQQWPPF-GUBZILKMSA-N Asp-Leu-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O DWOGMPWRQQWPPF-GUBZILKMSA-N 0.000 description 1
- BWJZSLQJNBSUPM-FXQIFTODSA-N Asp-Pro-Asn Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O BWJZSLQJNBSUPM-FXQIFTODSA-N 0.000 description 1
- KNOGLZBISUBTFW-QRTARXTBSA-N Asp-Trp-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C(C)C)C(O)=O KNOGLZBISUBTFW-QRTARXTBSA-N 0.000 description 1
- ZQFZEBRNAMXXJV-KKUMJFAQSA-N Asp-Tyr-His Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CC(=O)O)N)O ZQFZEBRNAMXXJV-KKUMJFAQSA-N 0.000 description 1
- 235000011299 Brassica oleracea var botrytis Nutrition 0.000 description 1
- 240000003259 Brassica oleracea var. botrytis Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 241000254173 Coleoptera Species 0.000 description 1
- ODDOYXKAHLKKQY-MMWGEVLESA-N Cys-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CS)N ODDOYXKAHLKKQY-MMWGEVLESA-N 0.000 description 1
- SRIRHERUAMYIOQ-CIUDSAMLSA-N Cys-Leu-Ser Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O SRIRHERUAMYIOQ-CIUDSAMLSA-N 0.000 description 1
- YAHZABJORDUQGO-NQXXGFSBSA-N D-ribulose 1,5-bisphosphate Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)C(=O)COP(O)(O)=O YAHZABJORDUQGO-NQXXGFSBSA-N 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 241000255925 Diptera Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- SSWAFVQFQWOJIJ-XIRDDKMYSA-N Gln-Arg-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(=O)N)N SSWAFVQFQWOJIJ-XIRDDKMYSA-N 0.000 description 1
- MGJMFSBEMSNYJL-AVGNSLFASA-N Gln-Asn-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O MGJMFSBEMSNYJL-AVGNSLFASA-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
- GNMQDOGFWYWPNM-LAEOZQHASA-N Gln-Gly-Ile Chemical compound CC[C@H](C)[C@H](NC(=O)CNC(=O)[C@@H](N)CCC(N)=O)C(O)=O GNMQDOGFWYWPNM-LAEOZQHASA-N 0.000 description 1
- ORYMMTRPKVTGSJ-XVKPBYJWSA-N Gln-Gly-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCC(N)=O ORYMMTRPKVTGSJ-XVKPBYJWSA-N 0.000 description 1
- ITZWDGBYBPUZRG-KBIXCLLPSA-N Gln-Ile-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O ITZWDGBYBPUZRG-KBIXCLLPSA-N 0.000 description 1
- CAXXTYYGFYTBPV-IUCAKERBSA-N Gln-Leu-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O CAXXTYYGFYTBPV-IUCAKERBSA-N 0.000 description 1
- SFAFZYYMAWOCIC-KKUMJFAQSA-N Gln-Phe-Arg Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N SFAFZYYMAWOCIC-KKUMJFAQSA-N 0.000 description 1
- KFHASAPTUOASQN-JYJNAYRXSA-N Gln-Phe-His Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CCC(=O)N)N KFHASAPTUOASQN-JYJNAYRXSA-N 0.000 description 1
- OSCLNNWLKKIQJM-WDSKDSINSA-N Gln-Ser-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O OSCLNNWLKKIQJM-WDSKDSINSA-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
- CVPXINNKRTZBMO-CIUDSAMLSA-N Glu-Arg-Asn Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)CN=C(N)N CVPXINNKRTZBMO-CIUDSAMLSA-N 0.000 description 1
- PVBBEKPHARMPHX-DCAQKATOSA-N Glu-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCC(O)=O PVBBEKPHARMPHX-DCAQKATOSA-N 0.000 description 1
- KRRFFAHEAOCBCQ-SIUGBPQLSA-N Glu-Ile-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KRRFFAHEAOCBCQ-SIUGBPQLSA-N 0.000 description 1
- WNRZUESNGGDCJX-JYJNAYRXSA-N Glu-Leu-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O WNRZUESNGGDCJX-JYJNAYRXSA-N 0.000 description 1
- AOCARQDSFTWWFT-DCAQKATOSA-N Glu-Met-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O AOCARQDSFTWWFT-DCAQKATOSA-N 0.000 description 1
- PMSMKNYRZCKVMC-DRZSPHRISA-N Glu-Phe-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CCC(=O)O)N PMSMKNYRZCKVMC-DRZSPHRISA-N 0.000 description 1
- WVWZIPOJECFDAG-AVGNSLFASA-N Glu-Phe-Cys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCC(=O)O)N WVWZIPOJECFDAG-AVGNSLFASA-N 0.000 description 1
- BPCLDCNZBUYGOD-BPUTZDHNSA-N Glu-Trp-Glu Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 BPCLDCNZBUYGOD-BPUTZDHNSA-N 0.000 description 1
- LZEUDRYSAZAJIO-AUTRQRHGSA-N Glu-Val-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LZEUDRYSAZAJIO-AUTRQRHGSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108010060309 Glucuronidase Proteins 0.000 description 1
- NZAFOTBEULLEQB-WDSKDSINSA-N Gly-Asn-Glu Chemical compound C(CC(=O)O)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)CN NZAFOTBEULLEQB-WDSKDSINSA-N 0.000 description 1
- JVACNFOPSUPDTK-QWRGUYRKSA-N Gly-Asn-Phe Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JVACNFOPSUPDTK-QWRGUYRKSA-N 0.000 description 1
- FMNHBTKMRFVGRO-FOHZUACHSA-N Gly-Asn-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)CN FMNHBTKMRFVGRO-FOHZUACHSA-N 0.000 description 1
- DHDOADIPGZTAHT-YUMQZZPRSA-N Gly-Glu-Arg Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DHDOADIPGZTAHT-YUMQZZPRSA-N 0.000 description 1
- LLZXNUUIBOALNY-QWRGUYRKSA-N Gly-Leu-Lys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN LLZXNUUIBOALNY-QWRGUYRKSA-N 0.000 description 1
- GAFKBWKVXNERFA-QWRGUYRKSA-N Gly-Phe-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 GAFKBWKVXNERFA-QWRGUYRKSA-N 0.000 description 1
- GGAPHLIUUTVYMX-QWRGUYRKSA-N Gly-Phe-Ser Chemical compound OC[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)C[NH3+])CC1=CC=CC=C1 GGAPHLIUUTVYMX-QWRGUYRKSA-N 0.000 description 1
- VDCRBJACQKOSMS-JSGCOSHPSA-N Gly-Phe-Val Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O VDCRBJACQKOSMS-JSGCOSHPSA-N 0.000 description 1
- JJGBXTYGTKWGAT-YUMQZZPRSA-N Gly-Pro-Glu Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O JJGBXTYGTKWGAT-YUMQZZPRSA-N 0.000 description 1
- NSVOVKWEKGEOQB-LURJTMIESA-N Gly-Pro-Gly Chemical compound NCC(=O)N1CCC[C@H]1C(=O)NCC(O)=O NSVOVKWEKGEOQB-LURJTMIESA-N 0.000 description 1
- IRJWAYCXIYUHQE-WHFBIAKZSA-N Gly-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)CN IRJWAYCXIYUHQE-WHFBIAKZSA-N 0.000 description 1
- VNNRLUNBJSWZPF-ZKWXMUAHSA-N Gly-Ser-Ile Chemical compound [H]NCC(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VNNRLUNBJSWZPF-ZKWXMUAHSA-N 0.000 description 1
- DBUNZBWUWCIELX-JHEQGTHGSA-N Gly-Thr-Glu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O DBUNZBWUWCIELX-JHEQGTHGSA-N 0.000 description 1
- FXTUGWXZTFMTIV-GJZGRUSLSA-N Gly-Trp-Arg Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)CN FXTUGWXZTFMTIV-GJZGRUSLSA-N 0.000 description 1
- 235000009438 Gossypium Nutrition 0.000 description 1
- 241000255967 Helicoverpa zea Species 0.000 description 1
- MJUUWJJEUOBDGW-IHRRRGAJSA-N His-Leu-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CN=CN1 MJUUWJJEUOBDGW-IHRRRGAJSA-N 0.000 description 1
- 101000654245 Homo sapiens Succinate dehydrogenase assembly factor 2, mitochondrial Proteins 0.000 description 1
- 241000209219 Hordeum Species 0.000 description 1
- GRRNUXAQVGOGFE-UHFFFAOYSA-N Hygromycin-B Natural products OC1C(NC)CC(N)C(O)C1OC1C2OC3(C(C(O)C(O)C(C(N)CO)O3)O)OC2C(O)C(CO)O1 GRRNUXAQVGOGFE-UHFFFAOYSA-N 0.000 description 1
- QTUSJASXLGLJSR-OSUNSFLBSA-N Ile-Arg-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N QTUSJASXLGLJSR-OSUNSFLBSA-N 0.000 description 1
- QYZYJFXHXYUZMZ-UGYAYLCHSA-N Ile-Asn-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N QYZYJFXHXYUZMZ-UGYAYLCHSA-N 0.000 description 1
- SCHZQZPYHBWYEQ-PEFMBERDSA-N Ile-Asn-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N SCHZQZPYHBWYEQ-PEFMBERDSA-N 0.000 description 1
- HDODQNPMSHDXJT-GHCJXIJMSA-N Ile-Asn-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O HDODQNPMSHDXJT-GHCJXIJMSA-N 0.000 description 1
- UDLAWRKOVFDKFL-PEFMBERDSA-N Ile-Asp-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N UDLAWRKOVFDKFL-PEFMBERDSA-N 0.000 description 1
- BGZIJZJBXRVBGJ-SXTJYALSSA-N Ile-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N BGZIJZJBXRVBGJ-SXTJYALSSA-N 0.000 description 1
- BALLIXFZYSECCF-QEWYBTABSA-N Ile-Gln-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N BALLIXFZYSECCF-QEWYBTABSA-N 0.000 description 1
- PNDMHTTXXPUQJH-RWRJDSDZSA-N Ile-Glu-Thr Chemical compound N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H]([C@H](O)C)C(=O)O PNDMHTTXXPUQJH-RWRJDSDZSA-N 0.000 description 1
- AREBLHSMLMRICD-PYJNHQTQSA-N Ile-His-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N AREBLHSMLMRICD-PYJNHQTQSA-N 0.000 description 1
- MASWXTFJVNRZPT-NAKRPEOUSA-N Ile-Met-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(=O)O)N MASWXTFJVNRZPT-NAKRPEOUSA-N 0.000 description 1
- IVXJIMGDOYRLQU-XUXIUFHCSA-N Ile-Pro-Leu Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O IVXJIMGDOYRLQU-XUXIUFHCSA-N 0.000 description 1
- FQYQMFCIJNWDQZ-CYDGBPFRSA-N Ile-Pro-Pro Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 FQYQMFCIJNWDQZ-CYDGBPFRSA-N 0.000 description 1
- CAHCWMVNBZJVAW-NAKRPEOUSA-N Ile-Pro-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)O)N CAHCWMVNBZJVAW-NAKRPEOUSA-N 0.000 description 1
- JHNJNTMTZHEDLJ-NAKRPEOUSA-N Ile-Ser-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O JHNJNTMTZHEDLJ-NAKRPEOUSA-N 0.000 description 1
- DTPGSUQHUMELQB-GVARAGBVSA-N Ile-Tyr-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 DTPGSUQHUMELQB-GVARAGBVSA-N 0.000 description 1
- NGKPIPCGMLWHBX-WZLNRYEVSA-N Ile-Tyr-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N NGKPIPCGMLWHBX-WZLNRYEVSA-N 0.000 description 1
- 108010025815 Kanamycin Kinase Proteins 0.000 description 1
- HGCNKOLVKRAVHD-UHFFFAOYSA-N L-Met-L-Phe Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 HGCNKOLVKRAVHD-UHFFFAOYSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 1
- KFKWRHQBZQICHA-STQMWFEESA-N L-leucyl-L-phenylalanine Natural products CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KFKWRHQBZQICHA-STQMWFEESA-N 0.000 description 1
- YKNBJXOJTURHCU-DCAQKATOSA-N Leu-Asp-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N YKNBJXOJTURHCU-DCAQKATOSA-N 0.000 description 1
- KTFHTMHHKXUYPW-ZPFDUUQYSA-N Leu-Asp-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KTFHTMHHKXUYPW-ZPFDUUQYSA-N 0.000 description 1
- RVVBWTWPNFDYBE-SRVKXCTJSA-N Leu-Glu-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RVVBWTWPNFDYBE-SRVKXCTJSA-N 0.000 description 1
- YVKSMSDXKMSIRX-GUBZILKMSA-N Leu-Glu-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O YVKSMSDXKMSIRX-GUBZILKMSA-N 0.000 description 1
- NEEOBPIXKWSBRF-IUCAKERBSA-N Leu-Glu-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O NEEOBPIXKWSBRF-IUCAKERBSA-N 0.000 description 1
- HYIFFZAQXPUEAU-QWRGUYRKSA-N Leu-Gly-Leu Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(C)C HYIFFZAQXPUEAU-QWRGUYRKSA-N 0.000 description 1
- VGPCJSXPPOQPBK-YUMQZZPRSA-N Leu-Gly-Ser Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O VGPCJSXPPOQPBK-YUMQZZPRSA-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
- QJXHMYMRGDOHRU-NHCYSSNCSA-N Leu-Ile-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O QJXHMYMRGDOHRU-NHCYSSNCSA-N 0.000 description 1
- JNDYEOUZBLOVOF-AVGNSLFASA-N Leu-Leu-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O JNDYEOUZBLOVOF-AVGNSLFASA-N 0.000 description 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
- PTRKPHUGYULXPU-KKUMJFAQSA-N Leu-Phe-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O PTRKPHUGYULXPU-KKUMJFAQSA-N 0.000 description 1
- 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
- IZPVWNSAVUQBGP-CIUDSAMLSA-N Leu-Ser-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IZPVWNSAVUQBGP-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
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 1
- LCNASHSOFMRYFO-WDCWCFNPSA-N Leu-Thr-Gln Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(N)=O LCNASHSOFMRYFO-WDCWCFNPSA-N 0.000 description 1
- GZRABTMNWJXFMH-UVOCVTCTSA-N Leu-Thr-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZRABTMNWJXFMH-UVOCVTCTSA-N 0.000 description 1
- VHTIZYYHIUHMCA-JYJNAYRXSA-N Leu-Tyr-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O VHTIZYYHIUHMCA-JYJNAYRXSA-N 0.000 description 1
- WFCKERTZVCQXKH-KBPBESRZSA-N Leu-Tyr-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O WFCKERTZVCQXKH-KBPBESRZSA-N 0.000 description 1
- MVJRBCJCRYGCKV-GVXVVHGQSA-N Leu-Val-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MVJRBCJCRYGCKV-GVXVVHGQSA-N 0.000 description 1
- AIMGJYMCTAABEN-GVXVVHGQSA-N Leu-Val-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIMGJYMCTAABEN-GVXVVHGQSA-N 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- IRNSXVOWSXSULE-DCAQKATOSA-N Lys-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN IRNSXVOWSXSULE-DCAQKATOSA-N 0.000 description 1
- UQRZFMQQXXJTTF-AVGNSLFASA-N Lys-Lys-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O UQRZFMQQXXJTTF-AVGNSLFASA-N 0.000 description 1
- NYTDJEZBAAFLLG-IHRRRGAJSA-N Lys-Val-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(O)=O NYTDJEZBAAFLLG-IHRRRGAJSA-N 0.000 description 1
- ZMYHJISLFYTQGK-FXQIFTODSA-N Met-Asp-Asn Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O ZMYHJISLFYTQGK-FXQIFTODSA-N 0.000 description 1
- MIXPUVSPPOWTCR-FXQIFTODSA-N Met-Ser-Ser Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MIXPUVSPPOWTCR-FXQIFTODSA-N 0.000 description 1
- 241001465754 Metazoa Species 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
- XUYPXLNMDZIRQH-LURJTMIESA-N N-acetyl-L-methionine Chemical compound CSCC[C@@H](C(O)=O)NC(C)=O XUYPXLNMDZIRQH-LURJTMIESA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- 108010034522 NNQQ peptide Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- NOFBJKKOPKJDCO-KKXDTOCCSA-N Phe-Ala-Tyr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O NOFBJKKOPKJDCO-KKXDTOCCSA-N 0.000 description 1
- SEPNOAFMZLLCEW-UBHSHLNASA-N Phe-Ala-Val Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)O SEPNOAFMZLLCEW-UBHSHLNASA-N 0.000 description 1
- HTTYNOXBBOWZTB-SRVKXCTJSA-N Phe-Asn-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N HTTYNOXBBOWZTB-SRVKXCTJSA-N 0.000 description 1
- CSYVXYQDIVCQNU-QWRGUYRKSA-N Phe-Asp-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O CSYVXYQDIVCQNU-QWRGUYRKSA-N 0.000 description 1
- NPLGQVKZFGJWAI-QWHCGFSZSA-N Phe-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O NPLGQVKZFGJWAI-QWHCGFSZSA-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
- YMIZSYUAZJSOFL-SRVKXCTJSA-N Phe-Ser-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O YMIZSYUAZJSOFL-SRVKXCTJSA-N 0.000 description 1
- ILGCZYGFYQLSDZ-KKUMJFAQSA-N Phe-Ser-His Chemical compound N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O ILGCZYGFYQLSDZ-KKUMJFAQSA-N 0.000 description 1
- MRWOVVNKSXXLRP-IHPCNDPISA-N Phe-Ser-Trp Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O MRWOVVNKSXXLRP-IHPCNDPISA-N 0.000 description 1
- BSKMOCNNLNDIMU-CDMKHQONSA-N Phe-Thr-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O BSKMOCNNLNDIMU-CDMKHQONSA-N 0.000 description 1
- KLYYKKGCPOGDPE-OEAJRASXSA-N Phe-Thr-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O KLYYKKGCPOGDPE-OEAJRASXSA-N 0.000 description 1
- BPIMVBKDLSBKIJ-FCLVOEFKSA-N Phe-Thr-Phe Chemical compound C([C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 BPIMVBKDLSBKIJ-FCLVOEFKSA-N 0.000 description 1
- YFXXRYFWJFQAFW-JHYOHUSXSA-N Phe-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N)O YFXXRYFWJFQAFW-JHYOHUSXSA-N 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 108700001094 Plant Genes Proteins 0.000 description 1
- 241000209504 Poaceae Species 0.000 description 1
- IWNOFCGBMSFTBC-CIUDSAMLSA-N Pro-Ala-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IWNOFCGBMSFTBC-CIUDSAMLSA-N 0.000 description 1
- ZCXQTRXYZOSGJR-FXQIFTODSA-N Pro-Asp-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O ZCXQTRXYZOSGJR-FXQIFTODSA-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
- CLNJSLSHKJECME-BQBZGAKWSA-N Pro-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H]1CCCN1 CLNJSLSHKJECME-BQBZGAKWSA-N 0.000 description 1
- VTFXTWDFPTWNJY-RHYQMDGZSA-N Pro-Leu-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VTFXTWDFPTWNJY-RHYQMDGZSA-N 0.000 description 1
- VGVCNKSUVSZEIE-IHRRRGAJSA-N Pro-Phe-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O VGVCNKSUVSZEIE-IHRRRGAJSA-N 0.000 description 1
- POQFNPILEQEODH-FXQIFTODSA-N Pro-Ser-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O POQFNPILEQEODH-FXQIFTODSA-N 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 240000000528 Ricinus communis Species 0.000 description 1
- 235000004443 Ricinus communis Nutrition 0.000 description 1
- SRTCFKGBYBZRHA-ACZMJKKPSA-N Ser-Ala-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SRTCFKGBYBZRHA-ACZMJKKPSA-N 0.000 description 1
- DWUIECHTAMYEFL-XVYDVKMFSA-N Ser-Ala-His Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 DWUIECHTAMYEFL-XVYDVKMFSA-N 0.000 description 1
- JPIDMRXXNMIVKY-VZFHVOOUSA-N Ser-Ala-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JPIDMRXXNMIVKY-VZFHVOOUSA-N 0.000 description 1
- OYEDZGNMSBZCIM-XGEHTFHBSA-N Ser-Arg-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OYEDZGNMSBZCIM-XGEHTFHBSA-N 0.000 description 1
- VAUMZJHYZQXZBQ-WHFBIAKZSA-N Ser-Asn-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O VAUMZJHYZQXZBQ-WHFBIAKZSA-N 0.000 description 1
- FIDMVVBUOCMMJG-CIUDSAMLSA-N Ser-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CO FIDMVVBUOCMMJG-CIUDSAMLSA-N 0.000 description 1
- DKKGAAJTDKHWOD-BIIVOSGPSA-N Ser-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CO)N)C(=O)O DKKGAAJTDKHWOD-BIIVOSGPSA-N 0.000 description 1
- RNMRYWZYFHHOEV-CIUDSAMLSA-N Ser-Gln-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RNMRYWZYFHHOEV-CIUDSAMLSA-N 0.000 description 1
- YPUSXTWURJANKF-KBIXCLLPSA-N Ser-Gln-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YPUSXTWURJANKF-KBIXCLLPSA-N 0.000 description 1
- DGPGKMKUNGKHPK-QEJZJMRPSA-N Ser-Gln-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CO)N DGPGKMKUNGKHPK-QEJZJMRPSA-N 0.000 description 1
- IXCHOHLPHNGFTJ-YUMQZZPRSA-N Ser-Gly-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CO)N IXCHOHLPHNGFTJ-YUMQZZPRSA-N 0.000 description 1
- SFTZWNJFZYOLBD-ZDLURKLDSA-N Ser-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO SFTZWNJFZYOLBD-ZDLURKLDSA-N 0.000 description 1
- HBTCFCHYALPXME-HTFCKZLJSA-N Ser-Ile-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HBTCFCHYALPXME-HTFCKZLJSA-N 0.000 description 1
- YUJLIIRMIAGMCQ-CIUDSAMLSA-N Ser-Leu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YUJLIIRMIAGMCQ-CIUDSAMLSA-N 0.000 description 1
- HEYZPTCCEIWHRO-IHRRRGAJSA-N Ser-Met-Phe Chemical compound OC[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 HEYZPTCCEIWHRO-IHRRRGAJSA-N 0.000 description 1
- RHAPJNVNWDBFQI-BQBZGAKWSA-N Ser-Pro-Gly Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O RHAPJNVNWDBFQI-BQBZGAKWSA-N 0.000 description 1
- CKDXFSPMIDSMGV-GUBZILKMSA-N Ser-Pro-Val Chemical compound [H]N[C@@H](CO)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O CKDXFSPMIDSMGV-GUBZILKMSA-N 0.000 description 1
- 102100031715 Succinate dehydrogenase assembly factor 2, mitochondrial Human genes 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
- 241000375392 Tana Species 0.000 description 1
- SKHPKKYKDYULDH-HJGDQZAQSA-N Thr-Asn-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O SKHPKKYKDYULDH-HJGDQZAQSA-N 0.000 description 1
- GARULAKWZGFIKC-RWRJDSDZSA-N Thr-Gln-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GARULAKWZGFIKC-RWRJDSDZSA-N 0.000 description 1
- AMXMBCAXAZUCFA-RHYQMDGZSA-N Thr-Leu-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AMXMBCAXAZUCFA-RHYQMDGZSA-N 0.000 description 1
- VRUFCJZQDACGLH-UVOCVTCTSA-N Thr-Leu-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VRUFCJZQDACGLH-UVOCVTCTSA-N 0.000 description 1
- IJVNLNRVDUTWDD-MEYUZBJRSA-N Thr-Leu-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IJVNLNRVDUTWDD-MEYUZBJRSA-N 0.000 description 1
- QHUWWSQZTFLXPQ-FJXKBIBVSA-N Thr-Met-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)NCC(O)=O QHUWWSQZTFLXPQ-FJXKBIBVSA-N 0.000 description 1
- NZRUWPIYECBYRK-HTUGSXCWSA-N Thr-Phe-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O NZRUWPIYECBYRK-HTUGSXCWSA-N 0.000 description 1
- NQQMWWVVGIXUOX-SVSWQMSJSA-N Thr-Ser-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NQQMWWVVGIXUOX-SVSWQMSJSA-N 0.000 description 1
- WPSKTVVMQCXPRO-BWBBJGPYSA-N Thr-Ser-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O WPSKTVVMQCXPRO-BWBBJGPYSA-N 0.000 description 1
- AKHDFZHUPGVFEJ-YEPSODPASA-N Thr-Val-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O AKHDFZHUPGVFEJ-YEPSODPASA-N 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- DNUJCLUFRGGSDJ-YLVFBTJISA-N Trp-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC1=CNC2=CC=CC=C21)N DNUJCLUFRGGSDJ-YLVFBTJISA-N 0.000 description 1
- GFZQWWDXJVGEMW-ULQDDVLXSA-N Tyr-Arg-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N)O GFZQWWDXJVGEMW-ULQDDVLXSA-N 0.000 description 1
- SGFIXFAHVWJKTD-KJEVXHAQSA-N Tyr-Arg-Thr 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@@H](C)O)C(O)=O SGFIXFAHVWJKTD-KJEVXHAQSA-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
- OEVJGIHPQOXYFE-SRVKXCTJSA-N Tyr-Asn-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O OEVJGIHPQOXYFE-SRVKXCTJSA-N 0.000 description 1
- DWJQKEZKLQCHKO-SRVKXCTJSA-N Tyr-Asn-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CS)C(=O)O)N)O DWJQKEZKLQCHKO-SRVKXCTJSA-N 0.000 description 1
- AOIZTZRWMSPPAY-KAOXEZKKSA-N Tyr-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)O AOIZTZRWMSPPAY-KAOXEZKKSA-N 0.000 description 1
- FZADUTOCSFDBRV-RNXOBYDBSA-N Tyr-Tyr-Trp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=C(O)C=C1 FZADUTOCSFDBRV-RNXOBYDBSA-N 0.000 description 1
- KLOZTPOXVVRVAQ-DZKIICNBSA-N Tyr-Val-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 KLOZTPOXVVRVAQ-DZKIICNBSA-N 0.000 description 1
- XLDYBRXERHITNH-QSFUFRPTSA-N Val-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)C(C)C XLDYBRXERHITNH-QSFUFRPTSA-N 0.000 description 1
- YODDULVCGFQRFZ-ZKWXMUAHSA-N Val-Asp-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O YODDULVCGFQRFZ-ZKWXMUAHSA-N 0.000 description 1
- UMPVMAYCLYMYGA-ONGXEEELSA-N Val-Leu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O UMPVMAYCLYMYGA-ONGXEEELSA-N 0.000 description 1
- NZGOVKLVQNOEKP-YDHLFZDLSA-N Val-Phe-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)N)C(=O)O)N NZGOVKLVQNOEKP-YDHLFZDLSA-N 0.000 description 1
- CKTMJBPRVQWPHU-JSGCOSHPSA-N Val-Phe-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)O)N CKTMJBPRVQWPHU-JSGCOSHPSA-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
- KSFXWENSJABBFI-ZKWXMUAHSA-N Val-Ser-Asn Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O KSFXWENSJABBFI-ZKWXMUAHSA-N 0.000 description 1
- RYHUIHUOYRNNIE-NRPADANISA-N Val-Ser-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N RYHUIHUOYRNNIE-NRPADANISA-N 0.000 description 1
- QHSSPPHOHJSTML-HOCLYGCPSA-N Val-Trp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)NCC(=O)O)N QHSSPPHOHJSTML-HOCLYGCPSA-N 0.000 description 1
- GTACFKZDQFTVAI-STECZYCISA-N Val-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC1=CC=C(O)C=C1 GTACFKZDQFTVAI-STECZYCISA-N 0.000 description 1
- AOILQMZPNLUXCM-AVGNSLFASA-N Val-Val-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN AOILQMZPNLUXCM-AVGNSLFASA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- QPMSXSBEVQLBIL-CZRHPSIPSA-N ac1mix0p Chemical compound C1=CC=C2N(C[C@H](C)CN(C)C)C3=CC(OC)=CC=C3SC2=C1.O([C@H]1[C@]2(OC)C=CC34C[C@@H]2[C@](C)(O)CCC)C2=C5[C@]41CCN(C)[C@@H]3CC5=CC=C2O QPMSXSBEVQLBIL-CZRHPSIPSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 108010009111 arginyl-glycyl-glutamic acid Proteins 0.000 description 1
- 108010069926 arginyl-glycyl-serine Proteins 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 108010010430 asparagine-proline-alanine Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 239000003139 biocide Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 239000003990 capacitor Substances 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 238000001311 chemical methods and process Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- 229930002868 chlorophyll a Natural products 0.000 description 1
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 1
- 229930002869 chlorophyll b Natural products 0.000 description 1
- NSMUHPMZFPKNMZ-VBYMZDBQSA-M chlorophyll b Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C=O)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 NSMUHPMZFPKNMZ-VBYMZDBQSA-M 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000012084 conversion product Substances 0.000 description 1
- 210000000877 corpus callosum Anatomy 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000005546 dideoxynucleotide Substances 0.000 description 1
- 108010054813 diprotin B Proteins 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 231100000284 endotoxic Toxicity 0.000 description 1
- 230000002346 endotoxic effect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 108010079413 glycyl-prolyl-glutamic acid Proteins 0.000 description 1
- 108010074027 glycyl-seryl-phenylalanine Proteins 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000012447 hatching Effects 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- GRRNUXAQVGOGFE-NZSRVPFOSA-N hygromycin B Chemical compound O[C@@H]1[C@@H](NC)C[C@@H](N)[C@H](O)[C@H]1O[C@H]1[C@H]2O[C@@]3([C@@H]([C@@H](O)[C@@H](O)[C@@H](C(N)CO)O3)O)O[C@H]2[C@@H](O)[C@@H](CO)O1 GRRNUXAQVGOGFE-NZSRVPFOSA-N 0.000 description 1
- 229940097277 hygromycin b Drugs 0.000 description 1
- 210000003000 inclusion body Anatomy 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002917 insecticide Substances 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 108010031424 isoleucyl-prolyl-proline Proteins 0.000 description 1
- 108010078274 isoleucylvaline Proteins 0.000 description 1
- 108010053037 kyotorphin Proteins 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010000761 leucylarginine Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 108010016686 methionyl-alanyl-serine Proteins 0.000 description 1
- 108010068488 methionylphenylalanine Proteins 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000001915 nurse cell Anatomy 0.000 description 1
- 230000031787 nutrient reservoir activity Effects 0.000 description 1
- 229940046166 oligodeoxynucleotide Drugs 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- PTMHPRAIXMAOOB-UHFFFAOYSA-N phosphoramidic acid Chemical compound NP(O)(O)=O PTMHPRAIXMAOOB-UHFFFAOYSA-N 0.000 description 1
- 229940061584 phosphoramidic acid Drugs 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 108010079317 prolyl-tyrosine Proteins 0.000 description 1
- 108010015796 prolylisoleucine Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000012723 sample buffer Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 230000028070 sporulation Effects 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000010023 transfer printing Methods 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- 108010038745 tryptophylglycine Proteins 0.000 description 1
- 108010020532 tyrosyl-proline Proteins 0.000 description 1
- IBIDRSSEHFLGSD-UHFFFAOYSA-N valinyl-arginine Natural products CC(C)C(N)C(=O)NC(C(O)=O)CCCN=C(N)N IBIDRSSEHFLGSD-UHFFFAOYSA-N 0.000 description 1
- 108010015385 valyl-prolyl-proline Proteins 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 235000020138 yakult Nutrition 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
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/32—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Bacillus (G)
- C07K14/325—Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
- A01H6/46—Gramineae or Poaceae, e.g. ryegrass, rice, wheat or maize
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Crystallography & Structural Chemistry (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Physiology (AREA)
- Botany (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明涉及一种编码杀虫剂蛋白的合成基因,所说的基因具有被修饰为其密码子用法与禾本科植物密码子用法一致的碱基顺序并以SEQ ID NO:1表示,一种含所说结构基因的表达载体和制备重组禾本科植物的方法,包括用载体转化来源于禾本科植物的原生质体,在存在水稻植物培养细胞的液体培养基中培养原生质体以形成集落,从所说集落再生植物。本发明还涉及具有昆虫抗性的重组禾本科植物。
Description
本发明涉及编码杀虫剂蛋白的合成基因、用该基因转化的重组禾本科植物用其制备方法。更具体地说,本发明涉及编码结晶蛋白(δ-内毒素)类似物的合成基因,所说结晶蛋白源于被称为杀虫剂细菌的苏云金芽胞杆菌库斯塔克(Kurustaki)HD-1变种,本发明还涉及具有昆虫和/或害虫抗性的禾本科植物的生产,包括用所说的合成基因转化特定的植物,并在合适的培养基中生长。
最近,越来越多的科研人员着手研究用杀虫细菌或细菌产物作为活性成份的细菌杀虫剂。例如,苏云金芽胞杆菌(下文称作B.t)就是一种已知能产生各种毒素如α-、β-和δ-内毒素的G+细菌,并对许多害虫具有高毒性(Angus T.A.,Nature 174;545,1954).在上述细菌性毒素中,由B.t在孢子形成过程中产生的δ-内毒素是作为蛋白质的结晶包涵体而引起兴趣的中心,并且近来被特别加以应用。已知不同的B.t.菌株能够产生具有适合作为杀虫剂活性成份特征的结晶蛋白质,所说的特性如窄的宿主谱,它使得结晶蛋白具有高度选择性、对动物包括人类无害以及环境可接受性。B.t.毒素的宿主谱包括属于鳞翅目、双翅目和鞘翅目的许多种昆虫。根据它们所产生的结晶蛋白的宿主谱不同,B.t.菌株分为5型。
在这类结晶蛋白的遗传工程方面的研究结果表明,通过编码毒性蛋白基因的转化来生产昆虫抗性植物成为可行的。例如,Vaeck等人,(Vaeck,et al,Nature 328:33-37,1987)(Plant Ge-netic Systems Inc,Belgium)已通过将一种编码源于B.t.的杀虫剂蛋白的基因整合到烟草基因组中而成功地制备了一种昆虫抗性烟草(Nicotiana)。根据上述报道,用所说基因S′端的1/2顺序获得的表达比用完整基因获得的表达更为有效。
Fischhoff等人(Bio/Technology 5:807-813,1987,Monsanto.Inc)成功地制备了用编码B.t.杀虫剂蛋白基因转化的重组西红柿。但是,上述两篇报道都没能用Western印迹检测到被转化基因的表达产物,这表明借助于天然存在的基因很难获得足够量的表达。
关于基因的翻译,有意义的是对组织间特定氨基酸变异的翻译活性。因此,与玉米胚中的tRNA相比,能过度合成玉米朊——一种富含谷氨酸、亮氨酸和丙氨酸的贮存蛋白的玉米胚乳中的tRNA更多地参予到编码这些氨基酸的基因翻译。这意味着特定组织中的tRNA以这种方式构建,以促进编码所需蛋白如玉米朊的基因的最适当的翻译,使得基因在所说组织中高效表达。
Wilbur等人(Plantphysiol.92:1-11,1990)指出了细菌和较高等植物如双子叶植物和单子叶植物在密码子使用方面的区别。因此,密码子XCG和XUA的使用在双子叶植物中为1.8%和3.2%,在单子叶植物中为6.3%和1.4%。密码子XXC和XXG的联合使用(下文称作密码子XXC/G的使用,其中,两个X的每一个分别选自A、G、C和T)在双子叶植物中为45%,在单子叶中为73.5%。已经明确,可被翻译的基因中的GC含量在单子叶植物,如禾本科水稻植物中比双子叶植物中高。对于细菌而言,密码子的使用根据菌株而变化。例如,密码子XCG、XUA和XXC/G在编码一种由B.t.库斯塔克HD-1(cryIA(b))变种产生的B.t.毒素——δ内毒素基因中的使用分别是10.4%、3.3%、和24.4%。这些事实说明.植物组织中的tRNA很难提供细菌基因的充分翻译。
最近,Prederick(Bio/Technology 8:939-942,1990)(Monsanto Inc.)首次报道了B.t.毒素可以在被修饰的基因转化的棉花(Gossypium)中高效表达,所说的修饰基因中,3′端的1/2顺序缺失且密码子发生了变化。因此,可用化学方法合成编码B.t.来源的δ-内毒素的基因,并连接到花椰菜花叶病病毒35S启动子的下游,所说的启动子含有35S启动子的增强子区。借助于土壤杆菌,将所得的上述基因用于转化。合成的基因可从苏云金芽胞杆菌库斯塔克HD-1变种(cryIA(b))和HD-73(cryIA(c))的基因制备。合成的基因提供B.t.毒素的高表达(约占叶中可溶性蛋白的0.05-0.1%),其表达率是用编码天然B.t.毒素基因获得的表达的约50-100倍,当用棉铃虫进行生物学测定来检测杀虫剂活性时,观察到约70-100%的保护性活性。
Frederic等人(Proc.Natl.Acad.Sci.USA,88:3324-3328,1991)试图在细菌基因上确定哪个区域因变动密码子进行修饰后能有效地在植物中表达所说的基因。他们选择了九个区进行修饰,并制备了在九个区的任何一个区或所有九个区进行修饰了的各种基因,然后用土壤杆菌将所制得的每种基因转化到烟草和西红柿植物中。当通过含有九个修饰区的基因获得的表达效率被估计为100%时,在5′末端700bp范围内含四个修饰区的基因显示出约80%的表达效率。相反,在用3′端的1/2顺序区被修饰的基因转化的植物中未检测到B.t.毒素。当基因所包括的修饰发生于5′末端246-283bp之间的区域时,表达效率最高,它是由含有九个修饰区基因所获得的表达效率的53-80%。
如上所述,在本发明之前,孟山都公司(Mon-Santo Inc)证明了可以通过改变密码子的用法达到细菌性蛋白(B.t.蛋白)在植物如棉花、烟草、西红柿中的高效表达。Lubrizol遗传学公司(Lubrizol Genetics Inc.)公开了一种编码杀虫剂结晶蛋白的合成基因,(日本专利,公开号:186989/1990),其中讨论了密码子的用法。但举证的植物仅是烟草。
从上所述可清楚看到,在本发明之前仅报道过细菌毒素在双子叶植物中的表达,没有报道说明在单子叶植物中表达出可检测量的杀虫剂结晶蛋白。
上述事实表明,经过缺失基因的3′端1/2顺序和使得密码子的用法与植物基因一致所进行的细菌基因的修饰,对于编码B.t.毒素的基因在植物中的表达是重要的。在这种情况下,本申请的发明人广泛研究和成功地制备了编码B.t.毒素类似物的合成基因,它首次能够在禾本科植物中高效表达。
因此,本发明提供了编码一种细菌性杀虫剂蛋白的合成基因,所说基因具有被修饰为其密码子的用法与禾本科植物基因相一致的碱基顺序,本发明还提供了含有所说基因的表达载体,用该载体转化的禾本科植物及其制备方法。
在整个说明书中,合成基因与待转化植物的基因两者间密码子用法的一致性以不同密码子的百分比表示。
本发明涉及的合成基因是在属于禾本科的植物中表达的,例如稻属、小麦属、大麦属,Secalu和Eshinochloa,优选的是稻属(水稻植物)。
本发明合成基因的制备是在编码源于苏云金芽胞杆菌库斯塔克变种HD-1(缩写成B.t.)的δ——内毒素的基因基础上进行的。
术语“重组植物”或“转化植物”指的是经转化作用而带有本发明合成基因,且能够对抗害虫的植物。
术语“B.t.毒素”指的是源于B.t.的毒性蛋白。在本说明书中,B.t.毒素不仅指由B.t.产生的天然存在的毒性蛋白,而且指由本发明合成基因编码的蛋白质,当其转化植物并进行表达时,则使植物具有抗昆虫特性。
术语“天然B.t.基因”指的是天然存在的编码B.t.毒素的基因。
本发明用化学和酶学方法制备基因,使之编码一种在功能上与天然B.t.毒素相当的蛋白质。该基因被设计为约200-400bp插入6bp限制性位点,以利于进一步的修饰作用。这些位点使得容易用其它位点来取代如决定宿主特异性的或类似的特性区。本发明基因的合成可通过将每个密码子的碱基成份改变成适合于植物而不影响由其编码的氨基酸来完成。进行修饰的目的是使天然B.t.基因的密码子用法适合于禾本科植物中基因密码子用法。确定适宜的密码子的用法将考虑下列密码子用法情况。
a)总的64个密码子中除了3个终止密码子(TAA、TGA和TAG)外的61个密码子的用法。
b)总的20种氨基酸中,除了仅由一个密码子编码的甲硫氨酸和色氨酸外的18种氨基酸的每一个编码密码子的用法。
C)编码上述18种氨基酸的密码子XXC/G的用法,在这种情况下,应考虑到单子叶植物与双子叶植物在密码子XXC/G用法方面的区别。
当希望本发明的合成基因将在特定的植物组织(如叶组织)中高效表达时,因为适宜的密码子用法在不同的组织间有所变化,因此重要的是使得所说合成基因的密码子用法与可能在叶组织中高效表达的基因的密码子用法一致。上述过程可通过以在叶组织中高效表达的基因的密码子用法时a)、b)、和c)为基础设计合成基因的碱基顺序来完成。
同样优选的是编码各个氨基酸的密码子应在整个合成基因中有一致的分布。
欲掺入到合成基因中的限制性位点(作为DNA片段的连接位点)选自存在于从B.t.库斯塔克HD-1(cryIA(b))变种天然蛋白的氨基酸顺序推测出的碱基顺序中的那些位点(J.Bac.166:801-811,1986)。如果所涉及的待转化植物是水稻植物(稻属)对碱基顺序进行修饰应考虑如下所确定的各种稻属组织中已知基因的密码子用法。
1)确定按d)、e)和f)所获得的水稻植物不同组织的基因中密码子用量的均值。
d)总的64个密码子中除3个终止密码子外的61个密码子的用量(图16);
e)总的20种氨基酸中除了由一个密码子编码的甲硫氨酸和色氨酸外的18种氨基酸的每种氨基酸的编码密码子的用法(图21,水稻);
f)编码上述18种氨基酸的密码子XXC/G的用法(图22,水稻)。
2)按下述方法确定可能在水稻植物的叶中高效表达的基因的密码子用法。
g)总的64个密码子中除3个终止密码子外的61个密码子的用法(图17);
h)总的20种氨基酸中除了由一个密码子编码的甲硫氨酸和色氨酸外的18种氨基酸的每种氨基酸的编码密码子的用法(图21,水稻叶);
i)编码上述18种氨基酸的密码子XXC/G的用法(图22,水稻叶)。
为了确定密码子,建议考虑下述方面。修饰编码B.t.δ-内毒素基因的碱基顺序,以使编码每一种氨基酸的密码子用法与在上文d)中(图16)获得的稻属(oriza)基因的密码子用法相符,其相符率为约±4%,更优选的是与在上文e)中(图21,水稻)获得的稻属(Oriza)基因的密码子用法相符,其相符率为40%。更优选的编码B.t.毒素中的优势氨基酸[在cryIA(b)中有10个氨基酸]的密码子用法应与上文f中(图22.水稻)获得的密码子相符,其相符率为±25%,所说的10种优势氨基酸包括精氨酸、亮氨酸、丝氨酸、甘氨酸、苏氨酸、缬氨酸、异亮氨酸、天冬氨酸、谷氨酸和苯丙氨酸。本文所用的术语“B.t.毒素中的优势氨基酸。”指的是在B.t.毒素中其含量排在前位的氨基酸。为了达到本发明目的,以使密码子用法尽可能地与在上文g)(图17)、h)(图21.水稻叶)和i)(图22.水稻叶)中得到的密码子用法一致这种方式选择密码子是最优选的。
如前文所述,希望编码各个氨基酸的密码子在整个合成基因中一致性分布。
因此而设计的一个作为例证的合成基因用SEQ ID NO:1表示。图18说明64个密码子中的61个密码子的用法(除去终止密码子),图21(BTHsyn)表示编码18种氨基酸(除去甲硫氨酸和色氨酸)中各个氨基酸的密码子用法,图22(BTHsyn)表示编码SEQ ID NO:1的合成基因的18种氨基酸的各种氨基酸的密码子XXC/G的用法。
有关合成基因与水稻植物天然基因间除终止密码子外的61个密码子用法的比较表明,所存在的差别除编码精氨酸的ACC为3.51%外,对其余所有密码子都为约2%。当在合成基因与可能在Oriza叶中高效表达的基因间进行比较时,密码子的用法变得更为一致。因此,对于编码精氨酸的AAC来说存在的差别最大(3.33%),而其它的密码子用法相当一致,这说明合成基因的密码子用法与在水稻植物叶中得到的基因的密码子用法更为相适应。
当对编码总的20种氨基酸中除甲硫氨酸和色氨酸外的18种氨基酸的密码子用法进行比较时,除了编码脯氨酸的CCG有36.9%的差别之外,几乎所有的密码子都表现出了相当好的一致性,有20%的差别。当对可能在Oriza叶中高效表达的基因进行比较时,密码子的用法变得更为一致。因此,对于编码赖氨酸的AAG来说差别最大(37%)而其它密码子的用法十分一致,这说明合成基因的密码子用法更好地适合了在水稻植物叶中得到基因的密码子用法。还对编码在B.t毒素中含量处于优势的10种氨基酸(精氨酸、亮氨酸、丝氨酸、甘氨酸、苏氨酸、缬氨酸、异亮氨酸、天冬氨酸、谷氨酸和苯丙氨酸)的密码子用法进行了比较。除了一种氨基酸(丝氨酸)差别为24.2%外,所有其它氨基酸中都表现出了相当好的一致性差别均为约15%。对在合成基因中和可能在Oriza叶组织中高效表达的基因中编码上述氨基酸的密码子XXC/G的用法进行了比较,结果表明除一个例外(丝氨酸为13.2%),前者与后者接近。
正如将为本领域技术人员显而易见的,借助于常规方法,如核甘酸的缺失、替代或增加能够很容易地对编码本发明杀虫剂蛋白的DNA进行修饰,产生具有相同活性或改善了活性的编码杀虫剂蛋白的DNA衍生物。因此,本发明还提供了这类通过常规方法获得的修饰基因。
图1:图解说明通过PCR(聚合酶链反应)合成双链DNA。
图2:图解说明通过PCR从寡核苷酸合成双链DNA。
图3:图解说明通过PCR从寡核苷酸合成双链DNA。
图4:用于PCR的引物(片段1)的碱基顺序。
图5:用于PCR的引物(片段5)的碱基顺序。
图6:用于PCR的引物(片段4)的碱基顺序。
图7:用于PCR的引物(片段2)的碱基顺序。
图8:用于PCR的引物(片段6)的碱基顺序。
图9:用于PCR的引物(片段3)的碱基顺序。
图10:在构建质粒pMCSC8、pMSCA8和pMCSK8时用于替代的克隆位点。
图11:图解说明质粒pBT1291的构建。
图12:将PCR产物进行电泳以筛选转化子时所得到的迁移图。
图13:通过Southern杂交检测用于转化的基因时所产生的迁移图。
图14:通过Northern杂交检测用于转化的基因的mRNA时所产生的迁移图。
图15:通过Western印迹检测在转基因的水稻叶中表达的B.t.毒素蛋白时所产生的迁移图。
图16:从水稻植物(Oriza)中分离的基因的密码子用法。
图17:在水稻植物叶中可能高效表达的基因的密码子用法。
图18:具有SEQIDNO:1碱基顺序的合成基因的密码子用法。
图19:转化子的第二代的PCR产物进行电泳时形成的迁移图。
图20:通过Western印迹检测在第二代(R1)植物叶中表达的B.t.毒素蛋白时产生的迁移图。
图21:在下列基因中观察到的编码各个氨基酸的密码子用法的组合:Oriza基因;在水稻植物叶中可能高效表达的基因;具有SEQIDNO:1碱基顺序的合成基因以及编码B.t.毒素中各个氨基酸的天然基因。
图22:在下列基因中观察到的编码各个氨基酸的密码子XXC/G用法的组合:Oriza基因;在水稻植物叶中可能高效表达的基因;具有SEQIDNO:1碱基顺序的合成基因以及编码B.t.毒素中各个氨基酸(除甲硫氨酸和色氨酸外)的天然基因。
可以借助任何常规方法制备如上所述设计的基因。例如,合成约20-100bp的单链寡核苷酸,并通过PCR将每个片段连接起来产生双链DNA(Am.J.Hum.Genet.37:172,1989),如下文及图1-3所示。方法A
此为在图1中图解说明的一种基本方法。根据该方法,通过合成一对70nt寡核苷酸,并且在用于PCR的标准反应混合物中进行反应,能获得所需要的双链DNA,所说的一对寡聚核苷酸中,每一段寡核苷酸在3′末端具有约10bp的互补顺序。方法B
此方法是基本方法A的一种应用,已在图2中说明。根据方法B,能够如下所述制备一段370bp双链DNA。用与上述方法A相同的方式处理寡核苷酸3和4,产生130bp双链DNA。通过PCR使所得的DNA与寡核苷酸2和5反应,再将所得的DNA与寡核苷酸将1和6一起进行PCR以生成所需要的双链DNA。该方法的优点在于容易获得限制性位点分开存在的长双链DNA。方法C:
该方法是基本方法A的另外一种应用,已在图3中说明。根据方法C,能如下所述制得含有如图3所示的限制性位点的260bp双链DNA。以与方法A相同的方式处理寡核苷酸对1和2以及3和4,产生130bp双链DNA。用对应于限制性位点(该处的片段是毗邻的)的限制性酶消化所得的DNA,继而用连接酶进行连接。将所得产物的一部分与寡核苷酸1和4再进行PCR以生成所需要的双链DNA。该方法所具有的优点在于不需要进行单个短片段的克隆,而且经酶学法连接的DNA的扩增能够迅速进行,因而利于所需DNA的克隆。
尽管这些方法适合于制备所需的DNA,但并不限于这些方法,并且也可以将方法B和方法C相结合使用。
然后将根据方法B和/或方法C制备的每一种双链DNA克隆到具有多克隆位点的适宜克隆载体中。适合的载体的例子包括那些从E.co-li衍生而来的载体,如pUC18,19(Gene33:103,1985)、pHSG298,299398,399(Gene,61:63,1987)和pBSIIKS+/-,pBSIIKS+/-(Stratagene Inc)它们从COLE1质粒及其类似物衍生而来,它们可以通过修饰这些已存在的载体的克隆性位点而获得。被克隆的片段的顺序可用Sanger等人(Proc.Natl.Acad.Sci.74:5463-5467,1977)的双脱氧法加以证实。两端具有限制性位点的单个DNA片断以相应的限制性酶进行消化,并在T4 DNA连接酶存在的条件下进行连接,产生编码B.t.毒素的完整长度的结构基因。
为了表达B.t.毒素,将结构基因插入到含有适合于结构基因在禾本科植物中表达的启动子和终止子以及内含子(如果需要的话)的质粒载体中。
启动子可以从那些已知在植物中具有功能的启动子中进行选择。这类启动子的例子包括来源于花椰菜花叶中病病毒的启动子,如CaMV35S(pB1221:EMBO.J.6:3901-3907,1987);rbcs(1,5-二磷酸核酮糖羧化酶);Cab(叶绿素a/b结合蛋白)(Science,244:174,1989)等。
终止子的例子包括那些来源于花椰菜花叶病病毒、NOS(蓝曙红(nopaline)合成酶)基因及其类似物的终止子,在结构基因和启动子之间含有内含子的载体同样可以用作高效表达载体。内含子的例子包括玉米Adhl(醇脱氢酶基因;Genes&Development 1:1183-1200,1987)的第一内含子,蓖麻油植物Cat(过氧化氢酶基因;Tanaka,etal,Nucleic Acids Research,18:6767-6770,1990)的第一内含子等。
为了达到本发明的目的,表达载体最好含有两个或多个选自潮霉素磷酸转移酶基因、新霉素磷酸转移酶基因、氯霉素乙酰转移酶基因、β-葡糖苷酸酶基因等的外源基因,并使所说外源基因之一充当选择性标记,以便可以筛选所需菌落,潮霉素磷酸酶基因是优选的选择性标记基因。
编码选择性标记物的外源基因和其它的基因可以包含于同一个表达载体中,或包含于在转化时同时应用的不同载体中。
本发明的载体用于转化禾本科植物。转化过程通常包括下列步骤。从特定的禾本科植物制备原生质体,并悬浮于液体培养基中。可以通过常规方法(如电脉冲处理)将表达载体转化到原生质体中。然后将所得的原生质体在含有禾本科培养细胞的培养基中孵育以获得细胞集落。筛选所需的集落,并进行生长培养以再生作为重组体产物的植物(Shimamoto et al,Hature,337:274-276,1989)。该过程将在下文详细说明。
可用任何目前存在的农用水稻种如NIPPONBARE、KOSHIHIKARI、SASANISHKI等作为植物来源以制备原生质体。在液体培养基中培养从植物组织,如成熟或未成熟的种子、叶鞘和/或根中制得的细胞悬液或胼胝体,并在0-50spm、25℃-30℃的条件下于含有能裂解细胞壁的酶(如纤维素酶)的溶液中处理3-16小时。在经酶处理后,将培养物过滤以分离未消化的物质。向滤液中加入2-5倍体积的KMC溶液(0.018M氯化钾,0.0817M氯化镁,0.085M氯化钙,PH6.0)(Theor.Appl.Genet,53:57-63,1978)等,然后离心得到纯化的原生质体。
转化过程的完成可通过例如,将1-100ug/ml携带上述制备的结构基因的表达载体与(2-10)×106/ml由植物制得的原生质体一起悬浮于液体培养基中(例如,含30-200mM氯化钙,0-50mM氯化镁和0.2-0.6M甘露糖醇的缓冲液),并施以电脉冲。最好根据日本专利公开号181791/1989所述,通过直流脉冲用100-1000μF电容器(起始电压:200-1000v/m,脉冲间隔:约1-50msec)进行电脉冲处理。
经脉冲处理过的原生质体悬浮于液体培养基中,如R2/MS培养基[R2培养基(Plant.Cell.Physiol,14:1113,1973)的无机成分与MS培养基(Murashige and Skoog,15:473-497,1962)的维生素混合物溶液或与MS培养基的混合物],所说的培养基最好含0.2-0.5%硝酸钾作为氮源。将悬液与含约1.0-3.0%琼脂糖的等量培养基(如R2/MS或MS培养基)混合,并在培氏平皿中迅即薄层铺展开以固化之。原生质体在固体培养基中的终浓度最好是约(5-50)×105/ml。将固化的琼脂切成约5-20mm大小的片段,并在液体培养基中以温和摇速(20-50rpm)于23-27℃下避光保温。
当原生质体来源于禾本科植物时,培养基最好含有约100-300mgPW/平皿的水稻植物培养细胞。或者,可用一个容器进行共同培养,容器的底部由滤膜形成或附着。因此,将含有原生质体的液体培养基置于所说容器中,再将容器交替浸泡于陪氏培养皿中含有水稻植物培养细胞的液体培养基中。被培养的细胞最好是以处于活性细胞分裂状态下的纯细胞群存在。通过将从水稻植物组织如种子、茎、根或花药制得的愈伤组织反复在液体培养基中进行亚培养,并筛选能快速分裂的细胞即可容易地获得上述培养细胞。
在孵育3-4周后,出现0.5-1mm的集落。如果载体含有还充当标记基因的外源基因,如潮霉素磷酸转移酶基因(hph),那么通过在培养开始后的第7-20天向培养基中加入10-100μg/ml的潮霉素并继续孵育即可容易地筛选出所需的转化子。被筛选出的集落转移至生长培养基中,并在光照(1000-4000lux)条件下于23-27℃孵育2-4周,获得约3-6mmφ的愈伤组织。生长培养基可以是补充了2ug/ml植物激素(如2,4-二氯苯氧基乙酸,2,4-D)和0.1-1.0%琼脂糖的琼脂培养基,如R2培养基。分离各个愈伤组织。当用于进行转化的载体含有hgh基因作为外源基因之一时,被分离的愈伤组织在补充了2.0-50μg/ml潮霉素的相同培养基中生长以证实潮霉素抗性的表达。
愈伤组织在含0.5-1.5%琼脂糖的R2/MS培养基中生长,所说培养基不含甲醛或补充了1-10mg.L的细胞分裂素、生长在光照(2000-4000lux)和23-27℃下进行2-10周,形成一个不定的芽或不定的胚,然后在无激素的R2,HS培养基中培养2-3周,生成能被移植的幼小植物。幼小植物在适宜的培养基(如蛭石等)中培养以获得所需要的转化植物,即重组水稻植物。
通过用任何已知的方法(例如Mol.Gen.Genet.211:172.1988)分离DNA,并将分离的DNA进行PCR(Am.j.Hum.Genet.37:172.1985)或Southern杂交(J.Mol.Biol.98:505.1980)可以证实本发明的合成基因存在于被转化的细胞或植物中。当转化细胞表达整合到植物基因组中的基因时,用被转化的基因作探针进行Morthern杂交(Thomas.P.et al.Proc.Natl.Acad.Sci.77:5201,1980)或用抗所说被转化基因的表达产物的抗血清进行Western印迹(Toxbin et al.Proc.Natl.Acad,Sci,76:4350,1979)来证实是有效的。
下面的实施例将进一步说明和详细公开本发明,但并不能认为是在限制本发明。
实施例1
编码B.t.毒素的结构基因的化学和酶学合成法及表达载体的构建
(1)寡脱氧核苷酸的制备
可以通过任何已知的标准方法(Matteucci et al,J.Am.Chem.Soc.103:3185-3192,1981:Beaucage et al.Tetrahedron lett,22:1859-1862,1981)合成用于构建编号B.t.毒素的结构基因(其碱基顺序以SEQ ID NO:1表示)的片段1-6的寡核苷酸。利用固相氨基磷酸三酯(phosphoramidite-triester)配对方法用Applied Biosystems391 DNA合成仪制备所有的寡核苷酸。将寡聚体去保护,并用28%氨水将它从固相载体上常规地分离出来,根据Hcbridg等人的方法(Biotechniques6:3662-367,1988)用寡核苷酸纯化柱(OPC柱Applied Biosystems)纯化粗寡核苷酸混合物。
(2)用PCR制备双链DNA
用于制备每一个片段的寡核苷酸的长度和顺序如图4-9所示,每个片段的限制性位点示于图11中。用上述方法B和C完成每个片段的制备。1)片段1的制备
片段1是在任一末端具有SalI和SpeI位点的190bp片段(图4)。根据方法B用4个寡核苷酸引物制备片段1。通过PCR(Am.J.Hum.Genet.37:172,1985)。使有意义引物1-3与反意引物1-4反应。PCR在含有总共100ul反应混合物[含每种引物10μM、10mMTris-HCL(PH8.3),1.5mM MgCl2,50mM KCl,0.005%吐温20,0.005%NP-40,0.001%明胶每种dATP,dGTP,dCTP和dTTP各200uM,5单位REPLITHERM热稳定性聚合酶(EPISENTRE Inc)]的DNA热循环仪PJ1000(PERKIN-ELMER CETUS Inc.)中进行。重复进行30次反应循环,每次包括94℃,1分钟;50-55℃,2分钟及72℃,3分钟。在方法B中,除了改变引物和DNA外,用于制备双链DNA的PCR通常在相同条件下进行。用5ul含120bpDNA的所得PCR混合物与有意义引物1-2和反意义引物1-4在同样条件下进行PCR,生成163bpDNA。再用5ul反应混合物与有意义引物1-1和反意义引物1-4进行PCR,生成所需的196bp双链DNA。
2)片段5的制备
片段5是在任一末端含EcoRI和XbaI位点的405bp片段(图5)。根据方法B,在上述1)中相同条件下以6个合成的寡核苷酸引物制备片段5。通过PCR使有意义引物5-3和反意义引物5-4在相似的条件下反应,生成145bpDNA。用5μl含所说145bpDNA的DNA溶液、有意义引物5-2和反意义引物5-5进行PCR,生成287bpDNA。用5μl含287bpDNA的溶液、有意义引物5-1和反意义引物5-6进行PCR,生成所需的411bp双链DNA。
3)片段4的制备
片段4是一个277bp片段,在任一末端带有EcoT141和ScaI位点并含有一个内部BglII位点(图6)。根据方法B和C,用5个合成的寡核苷酸引物制备片段4(图2和3)。通过PCR使有意义引物4-1和反意义引物4-2反应,生成具有EcoRT141和BglII末端的142bpDNA。将有意义引物4-4和反意义引物4-5进行PCR反应生成91bpDNA,继而再用5μl含91bpDNA的溶液、有意义引物4-3和反意义引物4-5进行PCR。生成具BglII和ScaI末端的151bpDNA。分别将142bpDNA和151bpDNA按如下所说进行处理。按常规方法用100μl酚/氯仿(1∶1)对含DNA的总反应物溶液进行脱蛋白处理,所得水层与10ul 3M NaOAC(PH5.2)和250μlEtOH混合,以沉淀DNA。用10单位限制性酶BglII在100μl反应溶液(含有50mM Tris-HCl PH7.5,10mM MgCl2,1mM二硫苏糖醇、100mMNaCl)中消化回收的DNA。反应混合物在1% SeakemGTG琼脂糖上(FMC Inc.)、IXTBE缓冲液中电脉,回收含DNA的条带,用SUPREC-01(Takara Inc.)从凝胶中纯化DNA。根据试剂盒所附手册说明,每种纯化的DNA的1/10用于连接反应,反应是用DNA连接试剂盒(Takara Inc.)在存在T4 DNA连接酶的50μl反应体系中进行。连接混合物(5μl)与有意义引物4-1及反意义引物4-5一起进行PCR,产生所需的283bp双链DNA。
4)片段2的制备
片段2是在两末端分别有ScaI和SacI位点并带一个内部StuI位点的414bp片段(图7)。根据上述方法B和C,用7个合成的寡核苷酸引物制备片段2。用有意义引物2-2与反意义引物2-3进行PCR,生成117bpDNA,继而与有意义引物2-1及反意义引物2-3进行PCR,产生具ScaI和StuI位点末端的179bpDNA。有意义引物2-5及反意义引物2-6进行PCR,产生129bpDNA,继而再与有意义引物2-4及反意义引物2-7进行PCR产生具StuI和SacI位点末端的254bpDNA。以上述3)相同的方式分别沉淀和回收179bpDNA和254bpDNA。用限制性酶StuI在100μl溶液中(10mM Tris-HCl PH7.5,10mM MgCl2,1mM二硫苏糖醇,50mM NaCl)消化回收的DNA,再按上述3)的方式进行纯化和连接。用连接混合物(5μl)与有意义引物2-1及反意义引物2-7进行PCR,产生所需的420bp双链DNA。
5)片段6的制备
片段6是在两末端具有SacI和HindIII位点的352bp片段(图8)。根据方法B(图2)以上述1)同样的方式用5个合成的寡核苷酸引物制备片段6。用有意义引物6-3和反意义引物6-4在相似条件下进行PCR产生146bpDNA。用5ul含所说146bpDNA的溶液与有意义引物6-2及反意义引物6-5进行PCR,产生284bpDNA。用5ul含284bpDNA的溶液与有意义引物6-1及反意义引物6-5进行PCR,产生所需的354bp双链DNA。
6)片段3的制备
片段3是在两末端具有HindIII位点和BclI位点并带一个内部PvuI位点的249bp片段(图9)。根据方法B和C(图2和3),如上文3)所述用5个合成的寡核苷酸引物制备片段3。用有意义引物3-1的反意义引物3-2进行PCR,产生具有HindIII的PvuI末端的117bpDNA。有意义引物3-4和反意义引物3-5进行PCR。产生88bpDNA,继而与有意义引物3-3及反意义引物3-5进行PCR,产生具PvuI和BclI末端的150bpDNA。以上述3)相同的方式分别沉淀179bpDNA和254bpDNA。用限制性酶PvuI在100μl溶液(含有20mM Tris-HCl PH8.5,10mMMgCl2,1mM二硫苏糖醇、100mM KCl)中消化回收的DNA,并以上述3)相同的方式进行纯化和连接。用连接混合物(5ul)与有意义引物3-1及反意义物3-5进行PCR,产生所需的255bpDNA双链DNA。
(3)编码B.t.毒素的全长结构基因的构建及含有该基因的表达载体
用上文(2)、3)所述的相似方式分别回收在上文(2)中获得的6个片段,并用对两末端的限制性位点有特异性的限制性酶消化,然后进行分离和纯化。
用DNA连接试剂盒(Takara Inc.)将上述所得片段克隆到下列克隆载体中,再用于转化E.coli DH5α。片段1:pBSIIKS+(Stratage-ne Inc.);片段2:来源于pHSG398(Gene 61:63,1987)的pMCSC8,它是通过在ECORI和HindIII克隆位点消化质粒pHSG398,并用如图10所示的优选克隆位点进行取代而得;片段3:来源于pUC18的pMCSA8,它是通过在ECORI和HindIII位点上消化质粒pUC18,并以如图10所示的优选克隆位点进行取代而得;片段4:来源于pHSG298(Gene,61:63,1987)的pMCSK8,它是通过在质粒pHSG298的EcoRI和HindIII克隆位点进行消化,并用如图10所示的优选克隆位点取代而得;片段5:pUC18(Gene,33:103,1985);片段6:pHSG399(Gene,61:63,1987)
通过确定插入的DNA的碱基顺序(图11)获得了分别克隆入了片段1-6的质粒pBTH1.经分离DNA并利用Sanger等人(Proc,Natl.Acad.Sci,74:5463-5467,1977)的双脱氧核苷酸测序法进行测序。
如图11所述进行质粒的构建。用10单位相应的限制性酶消化含有来源于花椰菜花叶病病毒的35S启动子及来源于蓖麻油植物的约1000bpSphI-Sall区内的过氧化氢酶基因内含子的高效表达载体pIG221(1μg,Tanaka et al,Nucleic Acids Research,18:6767-6770,1990),分离一个1000bp SpHI-Sall片段,继而亚克隆入质粒pHG398中,产生质粒pBTo1。
用限制性酶XbaI消化含片段1的质粒pBTH1,并用DNA纯端化试剂盒(Takara Inc.),根据试剂盒所附手册说明在含T4 DNA聚合酶的总体积20μl的反应体系中进行钝端化,继而用SpeI消化得到载体DNA。从质粒pBTO2中分离天然B.t.基因部分,pBTO2含有经限制酶SpeI和EcoRV消化而克隆入pBSIIkS+中的天然B.t.基因(源于pSY177,J.Bac-teriol,166:801-811,1986)的一部分(97bp SpeI-EcoRI片段)。用DNA连接试剂盒(Takara Inc.)对这些DNA片段,即载体DNA和上述制备的部分天然B.t.基因进行连接,产生质粒pBTH101。用限制性酶SalI和EcoRI消化质粒pBTH101,得到281bpDNA,继而连接至质粒pBTO1的SalI和EcoRI位点RI位点,产生质粒pBTH111。
在一平行实验中,根据如下所述构建质粒pBTH112。用限制性酶EcoRI和XbaI消化质粒pBTH5产生411bp片段,并以上述相同的方式连接至与XbaI有相同粘性末端的质粒pBTH4的限制性位点EcoRI和E-coT141,产生质粒pBTH102。用限制性酶ScaI和SacI消化质粒pBTH2产生414bpDNA片段,并以上述相同的方式连接到质粒pBTH102的限性位点ScaI和SacI上产生质粒pBTH112。
在另一个平行实验中,以上述相同方式将具有多个终止接头的约550bp片段与蓝曙红(nopaline)合成酶的约300bp多(A)额外信号顺序克隆到pMCSC8的BclI和KpnI位点,得到质粒pBT03,所说的终止接头含部分天然B.t,基因(pSY117,J.Bacteriol,166;801-811,1986)的238bpBclI-KpnI片段之间还包含有XbaI和SpeI位点。用HindIII和BclI限制性酶消化质粒pBTH03产生256bpDNA片段,继而以上述相同方式连接到pBT03的HindIII和BclI位点,产生质粒pBTH103。
用SacI和HindIII限制性酶消化质粒pBTH6产生352bpDNA,继而将以上述相同方式连接到质粒pBTH103和SacI的HindIII位点产生pBTH113。
最后,pBTH111的约1300bpSphI-EcoRI片段、pBTH112的约1100bpEcoRI-SacI片段和pBTH113于限制性位点SphI和SacI处连接,得到所需的全长度的合成B.t.基因和能够高效表达所说合成基因的表达载体pBT1291(图11)。
实施例2
编码B.t.毒素的合成基因在植物中的表达
(1)水稻植物原生质体的转化
表达载体pBT1291用于转化禾本科植物。一般说来,转化过程,包括:将源于禾本科植物的原生质体悬浮于液体培养基中,经电脉冲处理用携带合成基因的表达载体转化所说的原生质体、在含水稻植物培养细胞的适宜培养基中培养转化的原生质体以获得集落,并用已知方法(Shimamo-to et al,Nature.337:274-276,1989)再生来自所需集落的重组植物。
原生质体的制备方法如下。从源于农田水稻植物种(NIPPONBARE)的成熟胚愈伤组织制备细胞悬液,并生长3-5天,将悬液用含4%纤维素酶(Yakult Inc)、1%macerozymeR-10(YakultInc.)和0.4%甘露糖醇的酶液(pH5.6)于30℃处理3-4小时。当酶处理完毕,过滤培养物以分离未被消化的物质,所得滤液与4倍体积KMC溶液(0.118M KCl,0.0817MmgCl2,0.085MCaCl2,pH6.0,出处同上)结合,离心成为原生质体团丸,用KMC溶液洗2次。
所得的原生质体以8×106/ml的密度悬浮于缓冲液中(含70mMKCl5mM MgCl2 0.4M甘露糖醇0.1%MES pH5.8)。
向悬浮液中加入含合成的结构基因的质粒载体60μg/ml和质粒,如PGL2,(Nature 338:274-276,1989)60μg/ml),并冷却至5℃5分钟,所说质粒含有作为启动子的CaMv35S及作为外源基因的潮霉素磷酸转移酶基因和NOS(nopaline合成酶)或源于Camv的终止子基因。将混合物转移到无菌塑料槽中,并用平行电极(电容器:1000μF;起始电压500v/cm;脉冲间隔:约30msec)施以直流电脉冲。然后将悬液冷却至4℃10分钟,与等量R2/MS原生质体琼脂糖培养基(Mol.Gen.Gen-et.206:408,1987)混合,固化为约0.7mm厚度,得到固体琼脂糖(细胞密度约4×106/ml)。
将含有经电脉冲处理过的原生质体的固化琼脂糖切成约10mm大小的片,并置于含5mlR2/ms原生质体培养基的6cm陪氏平皿中。向平皿中加入约100mg(FW)水稻植物的培养细胞作为保护细胞(nurse cell)。将原生质体在29℃温和摇育(50rpm)培养约10天。
培养细胞的制备如下。将从由种子培育的水稻植物的根制得愈伤组织,每周在液体培养基中亚培养获得培养细胞的悬液,从悬液中筛选能活性分裂的发育好的细胞(1mmφ)。
培养10天后,用KMC溶液去掉保护细胞,继续孵育2-4天,加入20μg/ml潮霉素B,再孵育2-3周。
将一块琼脂糖块转移到含2mg/ml2,4-二氯苯氧乙酸(2,4-D)、6%葡萄糖和0.25%琼脂糖的R2软琼脂培养基中,所得混合物培养2-4周以使集落生长。分离增大的集落并转至R2软琼脂培养基上。
当所得的愈伤组织被转移到R2/MS再生培养基(3%山梨醇,2%蔗糖,1%琼脂糖,PH5.8)中,并在光照(2000-4000lux)、25℃条件下培养3-10周时,可观察到嫩枝和根的再生。当枝条长至约2cm时,可将其转移到含有R2/MS再生培养基的塑料合中,培养生长为幼小植物。然后进一步在蛭石盆中生长,得到成熟的重组水稻植物。
(2)通过PCR筛选转化子
如下所述,从潮霉素抗性集落中筛选被转化的合成基因,从部分潮霉素抗性集落中提取DNA(Mol.Gen.Genet.221:27,1988)。在含250ul再悬浮缓冲液(20mM Tris-HCl,10mM EDTA)的1.5ml微离心管中将两种愈伤组织匀浆化。向匀浆中加入20ul20%SDS,并在68℃培养15分钟。在加入150μl7.5M乙酸铵后,将混合物置于冰上30分钟,再于4℃下以15000rpm离心15分钟。向上清中加入1mlEtOH,在相同条件下离心,沉淀DNA。用70%EtOH洗DNA,干燥后溶于30μlTB缓冲液(10mM Tris-HCl.PH8.0,1mMEDTA)。
用图12所示的引物与所获得的上述DNA进行PCR以筛选转化基因。进行PCR的共50μl反应混合物包括5ul上述获得的DNA、每种引物各1μM、10mMTris-HCl(PH8.3),1.5mM MgCl2,50mM KCl,0.005%吐温20,0.005%NP-40、0.001%明胶、dATP、dGTP、dCTP和dTTP各200μM及5U REPLITHERM热稳定性DNA聚合酶(EPISENTRE Inc.)。PCR重复进行30-35次,每次反应循环包括94℃,1分钟;50℃,2分钟;72℃,3分钟,在DNA热循环仪PJ1000(PERKIN-ELMER CETUS Inc.)中进行,并用电泳对反应产物进行常规分析,正如图12所见,在质粒pBT1291转化的愈伤组织中观察到扩增DNA的1.0kb带。以同种方式筛选大量潮霉素抗性集落,证明47%的集落含有合成基因(质粒pBT1291)。
(3)全长度转化基因的检测
从上述(2)筛选获得的转化愈伤组织中提取DNA,用限制性酶XbaI20单位进行消化,并用Southern杂交分析(J.Mol.Biol.98:505.1980)用多引物DNA标记系统(Amersham Inc.,Feinberg et al.Analy-tical Biochem.137:266-267,1987)和x-32P-dcTP(Amersham Inc.,370MBq/ml 110TBq/mmol)标记PεT1291的XbaI片段,并用作探针。图13给出的结果表明在所有的转化物中检测到了推测为2.5kb的DNA带,它相当于B.t.合成基因与内含子的结合长度。
(4)作为转化基因转录本的mRNA的检测
从用Southern杂交(Analytical Biochem.162:156-159,1987)证实含有全长基因的转化愈伤组织中提取总RNA。通过Northern杂交(Thomas,P.et al.,Proc.Natl.Acad.Sci.,77:5201,1980)用按上述相同方法制得的探针对每-RNA(20μg)进行分析。图14给出的结果表明在所有实验对象中检测到了3.0-2.4kb的mRNA带,它对应于推测的转化基因的转录本。
(5)作为转化基因翻译产物的B.t.毒素的检测
从再生的水稻植物叶中提取SDS可溶性蛋白质,并用于Western印迹分析(Towbin et al,Proc.Natl.Acad.Sci.,76:4350,1979)中检测B.t.毒素。用液氮将一片约(2×0.5cm)重组水稻植物的叶冻干并在研钵中研成粉末,然后立即用100ul2倍样品缓冲液(25mMTris-HCl,PH6.8,4%SDS,8mMDTT,20%甘油,0.004%BPB,200μM白胃蛋白酶(leupepsin)抽提,提取物在沸水中加热5分钟,以12000rpm于4℃离心15分钟以分离作为样品溶液的上清液。用样品溶液进行SDS-PAGE电泳,并转移到Immobilon Membrane(MILLIPORE Inc.)上,印迹缓冲液为25mMTris,192mM甘氨酸,20%MeOH,转印电流200mA、电压35V,时间大于1小时。在B.t.毒素的检测中,用从被免疫兔子的抗B.t.毒素抗血清中纯化的免疫球蛋白G(IgG)作为第一抗体,用碱性磷酸酶结合羊抗兔IgG(JACKSON Inc.)作为第二抗体。
图15所示的结果表明检测到了能与从抗B.t.毒素抗血清中纯化的免疫球蛋白G(IgG)反应的分子量约72KD的蛋白质。估计该蛋白质的表达为SDS可溶性蛋白的约0.05%-0.1%。
(6)合成基因向子代R1的遗传其及表达
从高效表达B.t.毒素的水稻植物收集种子,播种并使其生长,用于检测合成基因向下一代(R1)的遗传和分离的比例。在播种14天后,以上述(2)相似的方式从植物的叶(约5×2mm)中提取DNA,并进行PCR。表1所示的结果表明该基因在三个独立的系中遗传,分离比约为3∶1。用电泳分析从C7-4系中提取的BNA(图19)。
表1
R1系 B.t..+ B.t.-
B2-9 98 28
C7-4 26 10
F2-11 17 6
以上述(5)相同的方式用约3月龄的F2-11系植物进行Weslcrn印迹分析,结果表明R1代植物与亲代植物表达的B.t.毒素量基本相同(图20)。
(7)重组植物的昆虫抗性试验
如下所述,检测水稻植物的重组植物对典型害虫(如稻纵卷叶螟,属鳞翅目)的抗性。在试验中,用被证实遗传了全长合成基因(B.t.-基因(+))的植物系,如B2-9系,(试验组)的14天龄幼苗检测接种给水稻植物的害虫的存活率(或死亡率)。
彻底冲洗14天龄B2-9系幼苗的根,并以用水浸湿的脱脂棉包裹,再用封口膜包裹,以防止水分蒸发,保持植物存活。将一株植物置于高约20cm的半透明塑料管中,并用2天龄的稻纵卷叶螟(2-3mm)进行接种。试验在25℃进行7天,其中调节昼夜间隔为12小时。在同一试验中,未处理的水稻植物(天然植物,HIPPUIIBARE)和已用因分离失去B.t.基因,(B.t.基因(一))的本发明表达载体以相同方式作为对照。结果如下表2和3所示。试验组(B.t.基因(+))的害虫存活率(或死亡率)明显区别于对照组(NIPPONBARE和B.t.基因(一))。也就是说,在试验组观察到的死亡率高于在对照组中所观察到的死亡率。试验组与对照组相对,前组害虫的存活率明显较低。
表2
稻纵卷叶螟的生长期
死亡2 3 4 5 6* 存活害虫的均重NIPPONBARE 1 1 4 8 8 7 3.75mgB.t.基因(-) 1 4 3 3.13mgB.t.基因(+)10 3 10 8 2.67mg
表3
稻纵卷叶螟的生长期
死亡 4* 存活害虫的均重NIPPONBARE 5 5 1.48mgB.t.基因(-) 4 4 1.58mgB.t.基因(+) 20 4 1.0mg*:每个数字代表时期(2-6),在其下面的数字代表各个时期的存活害虫数。
从上可知,表达了转化的编码B.t.毒素的基因的水稻能有效对抗属于鳞翅目的害虫。
总之,因为本发明的合成基因编码B.t.毒素,并具有与禾本科植物相适应的碱基顺序,它能在禾本科植物中高效表达,并使植物具有抗害虫抗性。本发明所得到的重组水稻植物可望应用于农业。
SEQ ID NO:1
顺序类型:对应于蛋白质的核苷酸
顺序长度:2172bp
股数:双
拓扑结构:线性
分子类型:其它核苷酸
合成DNA和基因组DNA的结合DNA
特征:1-2172BP肽ATG GAC AAC AAC CCC AAC ATC AAC GAA TGC ATC CCG TAC AAC TGC CTC 48Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu1 5 10 15AGC AAC CCG GAG GTC GAG GTC CTC GGC GGC GAG AGG ATC GAG ACT GGC 96Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly
20 25 30TAC ACC CCG ATC GAC ATC TCC CTC TCC CTC ACC CAG TTC CTC CTC AGC 144Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser
35 40 45GAG TTC GTC CCG GGC GCC GGC TTC GTC CTC GGA CTA GTT GAT ATA ATA 192Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile
50 55 60TGG GGA ATT TTT GGT CCC TCT CAA TGG GAC GCA TTT CTT GTA CAA ATT 240Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile65 70 75 80GAA CAG TTA ATT AAC CAA AGA ATA GAA GAA TTC GCC CGC AAC CAG GCC 288Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala
85 90 95ATC TCC AGG CTC GAG GGC CTC AGC AAC CTC TAC CAG ATC TAC GCC GAG 336Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu
100 105 110TCC TTC AGG GAG TGG GAG GCC GAC CCG ACC AAC CCG GCC CTC AGG GAG 384Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu
115 120 125GAG ATG CGC ATC CAG TTC AAC GAC ATG AAC AGC GCC CTC ACC ACC GCC 432Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala
130 135 140ATC CCG CTC TTC GCC GTC CAG AAC TAC CAG GTC CCG CTC CTC TCC GTC 480Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val145 150 155 160TAC GTC CAG GCC GCC AAC CTC CAC CTC TCC GTC CTC AGG GAC GTC TCC 528Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser
165 170 175GTG TTC GGC CAG AGG TGG GGC TTC GAC GCC GCG ACC ATC AAC AGC CGC 576Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg
180 185 190TAC AAC GAC TTG ACC AGG CTC ATC GGC AAC TAC ACC GAC CAC GCC GTC 624Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val
195 200 205CGC TGG TAC AAC ACC GGC CTC GAG CGC GTC TGG GGC CCG GAC TCT AGG 672Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg
210 215 220GAC TGG GTC AGG TAC AAC CAG TTC AGG AGG GAG CTG ACC CTC ACC GTC 720Asp Trp Val Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val225 230 235 240CTC GAC ATC GTC GCC CTC TTC TCC AAC TAC GAC AGC AGG ACC TAC CCG 768Leu Asp Ile Val Ala Leu Phe Ser Asn Tyr Asp Ser Arg Thr Tyr Pro
245 250 255ATC CGC ACC GTC TCC CAG CTC ACC AGG GAG ATC TAC ACC AAC CCG GTC 816Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val
260 265 270CTC GAG AAC TTC GAC GGC AGC TTC CGC GGC TCC GCC CAG GGC ATC GAG 864Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu
275 280 285GGC AGC ATC AGG AGC CCG CAC CTC ATG GAC ATC CTC AAC AGC ATC ACC 912Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr
290 295 300ATC TAC ACC GAC GCC CAC AGG GGC GAG TAC TAC TGG TCC GGC CAC CAG 960Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln305 310 315 320ATC ATG GCC TCC CCG GTC GGC TTC TCC GGC CCG GAG TTC ACC TTC CCG 1008Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro
325 330 335CTC TAC GGC ACG ATG GGC AAC GCC GCC CCG CAG CAA CGC ATC GTC GCC 1056Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala
340 345 350CAG CTC GGC CAG GGC GTC TAC AGG ACC CTC AGC TCC ACC CTC TAC AGG 1104Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg
355 360 365AGG CCT TTC AAC ATC GGC ATC AAC AAC CAG CAG CTC TCC GTC CTC GAC 1152Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp
370 375 380GGC ACC GAG TTC GCC TAC GGC ACC TCC TCC AAC TTG CCG TCC GCC GTC 1200Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val385 390 395 400TAC AGG AAG AGC GGC ACC GTG GAC TCC CTC GAC GAG ATC CCG CCG CAG 1248Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln
405 410 415AAC AAC AAC GTC CCG CCG AGG CAG GGC TTC AGC CAC CGC CTC AGC CAC 1296Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His
420 425 430GTC TCC ATG TTC CGC TCC GGC TTC AGC AAC AGC AGC GTC AGC ATC ATC 1344Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile
435 440 445AGA GCT CCC ATG TTC TCG TGG ATT CAC CGC TCG GCG GAG TTC AAC AAC 1392Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Glu Phe Asn Asn
450 455 460ATC ATC CCC TCG TCA CAG ATC ACG CAG ATC CCC CTG ACA AAG AGT ACG 1440Ile Ile Pro Ser Ser Gln Ile Thr Gln Ile Pro Leu Thr Lys Ser Thr465 470 475 480AAC CTG GGG TCG GGA ACA TCG GTG GTG AAG GGG CCC GGA TTC ACG GGG 1488Asn Leu Gly Ser Gly Thr Ser Val Val Lys Gly Pro Gly Phe Thr Gly
485 490 495GGA GAC ATC CTG CGC CGC ACT TCG CCC GGG CAG ATT TCA ACG CTG CGC 1536Gly Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln Ile Ser Thr Leu Arg
500 505 510GTG AAC ATC ACG GCG CCC CTG TCG CAG CGC TAT CGG GTG CGC ATT CGC 1584Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr Arg Val Arg Ile Arg
515 520 525TAC GCG TCT ACG ACA AAC CTT CAG TTC CAC ACG TCA ATC GAC GGG CGC 1632Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr Ser Ile Asp Gly Arg
530 535 540CCC ATC AAC CAG GGG AAC TTC TCG GCG ACA ATG TCG TCG GGG TCG AAC 1680Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr Met Ser Ser Gly Ser Asn545 550 555 560CTT CAG TCG GGA AGC TTC AGG ACC GTC GGC TTC ACC ACC CCG TTC AAC 1728Leu Gln Ser Gly Ser Phe Arg Thr Val Gly Phe Thr Thr Pro Phe Asn
565 570 575TTC TCC AAC GGC TCC AGC GTC TTC ACC CTC AGC GCT CAT GTC TTC AAC 1776Phe Ser Asn Gly Ser Ser Val Phe Thr Leu Ser Ala His Val Phe Asn
580 585 590TCC GGC AAC GAG GTC TAC ATC GAT CGC ATC GAG TTC GTC CCG GCC GAG 1824Ser Gly Asn Glu Val Tyr Ile Asp Arg Ile Glu Phe Val Pro Ala Glu
595 600 605GTC ACC TTC GAG GCC GAG TAC GAC CTC GAG AGG GCC CAG AAG GCC GTC 1872Val Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg Ala Gln Lys Ala Val
610 615 620AAC GAG CTG TTC ACC TCC AGC AAC CAG ATC GGC CTC AAG ACC GAC GTC 1920Asn Glu Leu Phe Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val625 630 635 640ACC GAC TAC CAC ATT GAT CAA GTA TCC AAT TTA GTT GAG TGT TTA TCT 1968Thr Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Glu Cys Leu Ser
645 650 655GAT GAA TTT TGT CTG GAT GAA AAA AAA GAA TTG TCC GAG AAA GTC AAA 2016Asp Glu Phe Cys Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys
660 665 670CAT GCG AAG CGA CTT AGT GAT GAA CGG AAT TTA CTT CAA GAT CCA AAC 2064His Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn
675 680 685TTT AGA GGG ATC AAT AGA CAA CTA GAC CGT GGC TGG AGA GGA AAT ACG 2112Phe Arg Gly Ile Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Asn Thr
690 695 700GAT ATT ACC ATC CAA GGA GGC CAT GAC GTA TTC AAA GAG AAT TAC GTT 2160Asp Ile Thr Ile Gln Gly Gly His Asp Val Phe Lys Glu Asn Tyr Val705 710 715 720ACG CTA TTG GGT 2172Thr Leu Leu GlySEQ ID NO:2顺序长度:51bp顺序类型:核酸股数:单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)TTGGTCGACC CATGGACAAC AACCCCAACA TCAACGAATGCATCCCGTAC A 51SEQ ID NO:3顺序长度:71bp顺序类型:核酸股数:单拓扑结构:线性分子类型:其它核苷酸(用于PCR的合成DNA)ACGAATGCAT CCCGTACAAC TGCCTCAGCA ACCCGGAGGTCGAGGTCCTC GGCGGCGAGA 60GGATCGAGAC T 71SEQ ID NO:4顺序长度:42bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GATCGAGACT GGCTACACCC CGATCGACAT CTCCCTCTCCCT 42SEQ ID NO:5顺序长度:70bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)TCAACTAGTC CGAGGACGAA GCCGGCGCCC GGGACGAACTCGCTGAGGAG GAACTGGGTG 60AGGGAGAGGG 70SEQ ID NO:6顺序长度:70bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GAAGAATTCG CCCGCAACCA GGCCATCTCC AGGCTCGAGGGCCTCAGCAA CCTCTACCAG 60ATCTACGCCG 70SEQ ID NO:7顺序长度:79bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)CTACGCCGAG TCCTTCAGGG AGTGGGAGGC CGACCCGACCAACCCGGCCC TCAGGGAGGA 60GATGCGCATC CAGTTCAAC 79SEQ ID NO:8顺序长度:77bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)AGTTCAACGA CATGAACAGC GCCCTCACCA CCGCCATCCCGCTCTTCGCC GTCCAGAACT 60ACCAGGTCCC GCTCCTC 77SEQ ID NO:9顺序长度:77bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)CTCTGGCCGA ACACGGAGAC GTCCCTGAGG ACGGAGAGGTGGAGGTTGGC GGCCTGGACG 60TAGACGGAGA GGAGCGG 77SEQ ID NO:10顺序长度:79bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)CGGTGTAGTT GCCGATGAGC CTGGTCAAGT CGTTGTAGCGGCTGTTGATG GTCGCGGCGT 60CGAAGCCCCA CCTCTGGCC 79SEQ ID NO:11顺序长度:70bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GTCTCTAGAG TCCGGGCCCC AGACGCGCTC GAGGCCGGTGTTGTACCAGC GGACGGCGTG 60GTCGGTGTAG 70SEQ ID NO:12顺序长度:73bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GACCCTAGGG ACTGGGTCAG GTACAACCAG TTCAGGAGGGAGCTGACCCT CACCGTCCTC 60GACATCGTCG CCC 73SEQ ID NO:13顺序长度:79bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)TGTAGATCTC CCTGGTGAGC TGGGAGACGG TGCGGATCGGGTAGGTCCTG CTGTCGTAGT 60TGGAGAAGAG GGCGACGAT 79SEQ ID NO:14顺序长度:68bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GAGATCTAGA CCAACCCGGT CCTCGAGAAC TTCGACGGCAGCTTCCGCGG CTCCGCCCAG 60GGCATCGA 68SEQ ID NO:15顺序长度:29bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GGCATCGAGG GCAGCATCAG GAGCCCCGCA 29SEQ ID NO:16顺序长度:70bp顺序类型:核酸股数:单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)AGTAGTACTC GCCCCTGTGG GCGTCGGTGT AGATGGTGATGCTGTTGAGG ATGTCCATGA 60GGTGCGGGCT 70SEQ ID NO:17顺序长度:70bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GCGAGTACTA CTGGTCCGGC CACCAGATCA TGGCCTCCCCGGTCGGCTTC TCCGGCCCGG 60AGTTCACCTT 70SEQ ID NO:18顺序长度:54bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)TTCACCTTCC CGCTCTACGG CACGATGGGC AACGCCGCCCCGCAGCAACG CATC 54SEQ ID NO:19顺序长度:71bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GAAAGGCCTC CTGTAGAGGG TGGAGCTGAG GGTCCTGTAGACGCCCTGGC CGAGCTGGGC 60GACGATGCGT T 71SEQ ID NO:20顺序长度:71bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)CAGGAGGCCT TTCAACATCG GCATCAACAA CCAGCAGCTCTCCGTCCTCG ACGGCACCGA 60GTTCGCCTAC G 71SEQ ID NO:21顺序长度:69bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)CGCCTACGGC ACCTCCTCCA ACTTGCCGTC CGCCGTCTACAGGAAGAGCG GCACCGTGGA 60CTCCCTCGA 69SEQ ID NO:22顺序长度:69bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)CTGAGGCGGT GGCTGAAGCC CTGCCTCGGC GGGACGTTGTTGTTCTGCGG CGGGATCTCG 60TCGAGGGAG 69SEQ ID NO:23顺序长度:70bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)TGGGAGCTCT GATGATGCTG ACGCTGCTGT TGCTGAAGCCGGAGCGGAAC ATGGAGACGT 60GGCTGAGGCG 70SEQ ID NO:24顺序长度:80bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)AGAGCTCCCA TGTTCTCGTG GATTCACCGC TCGGCGGAGTTCAACAACAT CATCCCCTCG 60TCACAGATCA CGCAGATCCC 80SEQ ID NO:25顺序长度:78bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)CGCAGATCCC CCTGACAAAG AGTACGAACC TGGGGTCGGGAACATCGGTG GTGAAGGGGC 60CCGGATTCAC GGGGGGAG 78SEQ ID NO:26顺序长度:78bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)ACGGGGGGAG ACATCCTGCG CCGCACTTCG CCCGGGCAGATTTCAACGCT GCGCGTGAAC 60ATCACGGCGC CCCTGTCG 78SEQ ID NO:27顺序长度:78bp顺序类型:核酸股数:单拓扑结构:线性分子类型:其它核酸(用于PCR的合成)CCGTCGATTG ACGTGTGGAA CTGAAGGTTT GTCGTAGACGCGTAGCGAAT GCGCACCCGA 60TAGCGCTGCG ACAGGGGC 78SEQ ID NO:28顺序长度:80bp顺序类型:核酸股数:单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GAAGCTTCCC GACTGAAGGT TCGACCCCGA CGACATTGTCGCCGAGAAGT TCCCCTGGTT 60GATGGGGCGC CCGTCGATTG 80SEQ ID NO:29顺序长度:63bp顺序类型:核酸股数:单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GGGAAGCTTC AGGACCGTCG GCTTCACCAC CCCGTTCAACTTCTCCAACG GCTCCAGCGT 60CTT 63SEQ ID NO:30顺序长度:63bp顺序类型:核酸股 数:单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)ATGCGATCGA TGTAGACCTC GTTGCCGGAG TTGAAGACATGAGCGCTGAG GGTGAAGACG 60CTG 63SEQ ID NO:31顺序长度:70bp顺序类型:核酸股数:单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)CATCGATCGC ATCGAGTTCG TCCCGGCCGA GGTCACCTTCGAGGCCGAGT ACGACCTCGA 60GAGGGCCCAG 70SEQ ID NO:32顺序长度:26bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)GGGCCCAGAA GGCCGTCAAC GAGCTG 26SEQ ID NO:33顺序长度:70bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)ACTTGATCAA TGTGGTAGTC GGTGACGTCG GTCTTGAGGCCGATCTGGTT GCTGGAGGTG 60AACAGCTCGT 70SEQ ID NO:34顺序长度:58bP顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)AATTGCATGC ATGCATGAAT TCCCTAGGAG TACTGAGCTCAAGCTTTGAT CAGGTACC 58SEQ ID NO:35顺序长度:58bp顺序类型:核酸股数: 单拓扑结构:线性分子类型:其它核酸(用于PCR的合成DNA)AGCTGGTACC TGATCAAAGC TTGAGCTCAG TACTCCTAGGGAATTCATGC ATGCATGC 58
Claims (5)
1.一种制备编码杀虫剂蛋白的合成基因的方法,其中所说的基因具有被修饰为其密码子用法与禾本科植物基因的密码子用法相一致的碱基顺序,其特征在于,所述方法是用化学和酶学方法完成的。
2.权利要求1所要求的方法,其中所说的基因被合成为使之密码子的用法与禾本科植物基因中的密码子用法相一致,其差别在约±4%以内。
3.权利要求1所要求的方法,其中所说的基因被合成为使其密码子用法与禾本科植物中编码各个氨基酸的基因的密码子用法相一致,其差别在±40%以内。
4.权利要求1所要求的方法,其中所说的基因被合成为使其编码天然存在的杀虫剂蛋白中的10种优势氨基酸的密码子用法与编码组成禾本科植物蛋白质的各个氨基酸的密码子XXC和XXG的联合密码子用法相一致,其差别为±25%,在所说的联合密码子用法中,两个X中的每一个都分别选自A、G、C和T。
5.权利要求1所要求的方法,其中所说的基因由SEQ ID No:1表示。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP34005991 | 1991-11-29 | ||
JP340059/1991 | 1991-11-29 | ||
JP340059/91 | 1991-11-29 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1073717A CN1073717A (zh) | 1993-06-30 |
CN1056881C true CN1056881C (zh) | 2000-09-27 |
Family
ID=18333331
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN92115178A Expired - Fee Related CN1056881C (zh) | 1991-11-29 | 1992-11-28 | 编码杀虫剂蛋白的基因、用该基因转化的禾本科植物及其制备方 |
Country Status (4)
Country | Link |
---|---|
US (1) | US5436391A (zh) |
KR (1) | KR100217517B1 (zh) |
CN (1) | CN1056881C (zh) |
TW (1) | TW261517B (zh) |
Families Citing this family (341)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6403865B1 (en) | 1990-08-24 | 2002-06-11 | Syngenta Investment Corp. | Method of producing transgenic maize using direct transformation of commercially important genotypes |
UA48104C2 (uk) * | 1991-10-04 | 2002-08-15 | Новартіс Аг | Фрагмент днк, який містить послідовність,що кодує інсектицидний протеїн, оптимізовану для кукурудзи,фрагмент днк, який забезпечує направлену бажану для серцевини стебла експресію зв'язаного з нею структурного гена в рослині, фрагмент днк, який забезпечує специфічну для пилку експресію зв`язаного з нею структурного гена в рослині, рекомбінантна молекула днк, спосіб одержання оптимізованої для кукурудзи кодуючої послідовності інсектицидного протеїну, спосіб захисту рослин кукурудзи щонайменше від однієї комахи-шкідника |
EP0793717B1 (en) * | 1994-10-24 | 2005-01-26 | The Texas A & M University System | Oral immunization with transgenic plants |
WO1997013843A1 (en) * | 1995-10-12 | 1997-04-17 | Cornell Research Foundation, Inc. | Production of water stress or salt stress tolerant transgenic cereal plants |
WO1999007210A1 (en) | 1997-08-12 | 1999-02-18 | North Carolina State University | Genetically engineered duckweed |
US6121521A (en) * | 1998-04-01 | 2000-09-19 | Novartis Ag | Chimeric insecticidal protein and DNA coding therefor |
US6399857B1 (en) | 1999-09-22 | 2002-06-04 | Paradigm Genetics, Inc. | Modified nucleotide sequence encoding a LexA DNA binding domain optimized for arabidopsis species |
WO2002010190A2 (en) * | 2000-07-29 | 2002-02-07 | Genaissance Pharmaceuticals, Inc. | Haplotypes of the il12b gene |
US20030033626A1 (en) | 2000-07-31 | 2003-02-13 | Hahn Frederick M. | Manipulation of genes of the mevalonate and isoprenoid pathways to create novel traits in transgenic organisms |
IL154596A0 (en) | 2000-08-25 | 2003-09-17 | Basf Plant Science Gmbh | Plant polynucleotides encoding novel prenyl proteases |
US6875907B2 (en) | 2000-09-13 | 2005-04-05 | Pioneer Hi-Bred International, Inc. | Antimicrobial peptides and methods of use |
MXPA03011890A (es) | 2001-06-22 | 2004-06-03 | Du Pont | Polinucleotidos de defensina y metodos de uso. |
WO2003020914A2 (en) | 2001-09-05 | 2003-03-13 | Basf Plant Science Gmbh | Protein phosphatase stress-related polypeptides and methods of use in plants |
US7220585B2 (en) | 2001-11-09 | 2007-05-22 | Basf Plant Science Gmbh | Transcription factor stress-related polypeptides and methods of use in plants |
EP2329708B1 (en) | 2002-07-10 | 2016-10-19 | The Department of Agriculture, Western Australia | Wheat plants having increased resistance to imidazolinone herbicides |
CA2492917C (en) | 2002-07-19 | 2011-10-18 | University Of South Carolina | Compositions and methods for the modulation of gene expression in plants |
US7460252B2 (en) * | 2003-01-13 | 2008-12-02 | Axiohm Transaction Solutions, Inc. | Graphical printing system and method using text triggers |
CN102586285B (zh) | 2003-02-20 | 2016-01-13 | 阿则耐克斯公司 | δ-内毒素基因及其使用方法 |
EP2194135A3 (en) | 2003-04-15 | 2010-10-06 | BASF Plant Science GmbH | Nucleic acid sequences from yeast encoding proteins associated with abiotic stress response and transformed plant cells and plants with increased tolerance to environmental stress |
US9382526B2 (en) | 2003-05-28 | 2016-07-05 | Basf Aktiengesellschaft | Wheat plants having increased tolerance to imidazolinone herbicides |
EP2982240B1 (en) | 2003-08-29 | 2019-07-31 | Instituto Nacional de Tecnologia Agropecuaria | Rice plants having increased tolerance to imidazolinone herbicides |
US20060260004A1 (en) * | 2004-04-01 | 2006-11-16 | Yiwen Fang | Par-related protein promoters |
BRPI0512904A (pt) | 2004-06-30 | 2008-04-15 | Pioneer Hi Bred Int | método para o aumento da resistência de plantas a patógenos de fungo |
BRPI0512920B1 (pt) | 2004-07-02 | 2017-05-16 | Du Pont | cassete de expressão, microorganismo transformado, método para indução de resistência a patógeno de planta em uma planta, composição anti-patogênica e método para proteção de uma planta contra um patógeno de planta |
EP2053057A3 (en) | 2004-09-24 | 2009-07-15 | BASF Plant Science GmbH | Nucleic acid sequences encoding proteins associated with abiotic stress response and plant cells and plants with increased tolerance to environmantal stress |
EP1831376A1 (en) | 2004-12-28 | 2007-09-12 | Pioneer Hi-Bred International, Inc. | Improved grain quality through altered expression of seed proteins |
ES2692594T1 (es) | 2005-03-02 | 2018-12-04 | Instituto Nacional De Tecnologia Agropecuaria | Plantas de arroz resistentes a herbicidas, polinucleótidos que codifican proteínas de la subunidad grande de la acetohidroxiácido sintasa resistentes a herbicidas y métodos para su uso |
EP2130839B1 (en) | 2005-04-01 | 2012-03-07 | Athenix Corporation | AXMI-036 a delta-endotoxin gene and methods for its use |
AU2006235263B2 (en) | 2005-04-08 | 2012-05-24 | BASF Agricultural Solutions Seed US LLC | Identification of a new class of EPSP synthases |
CA2612016A1 (en) | 2005-06-17 | 2006-12-21 | Basf Plant Science Gmbh | Lecitin-like protein kinase stress-related polypeptides and methods of use in plants |
US7598367B2 (en) * | 2005-06-30 | 2009-10-06 | Ceres, Inc. | Early light-induced protein promoters |
CA3006126C (en) | 2005-07-01 | 2022-04-19 | Basf Se | Herbicide-resistant sunflower plants, polynucleotides encoding herbicide-resistant acetohydroxy acid synthase large subunit proteins, and methods of use |
AU2006270158C1 (en) | 2005-07-18 | 2012-03-29 | Basf Plant Science Gmbh | Yield increase in plants overexpressing the SHSRP genes |
WO2007011625A2 (en) | 2005-07-18 | 2007-01-25 | Basf Plant Science Gmbh | Yield increase in plants overexpressing the accdp genes |
WO2007020198A2 (en) | 2005-08-12 | 2007-02-22 | Basf Plant Science Gmbh | Nucleic acid sequences encoding proteins associated with abiotic stress response and plant cells and plants with increased tolerance to environmental stress |
EP1945776A2 (en) | 2005-11-10 | 2008-07-23 | Pioneer Hi-Bred International, Inc. | Dof (dna binding with one finger) sequences and methods of use |
US20070199096A1 (en) | 2005-11-14 | 2007-08-23 | E.I. Du Pont De Nemours And Company | Compositions and Methods for Altering Alpha- and Beta-Tocotrienol Content |
AR057205A1 (es) | 2005-12-01 | 2007-11-21 | Athenix Corp | Genes grg23 y grg51 que confieren resistencia a herbicidas |
US20080313769A9 (en) | 2006-01-12 | 2008-12-18 | Athenix Corporation | EPSP synthase domains conferring glyphosate resistance |
US7622573B2 (en) | 2006-01-17 | 2009-11-24 | Biolex, Inc. | Expression control elements from the lemnaceae family |
WO2007100897A2 (en) * | 2006-02-27 | 2007-09-07 | Edenspace System Corporation | Energy crops for improved biofuel feedstocks |
AR059724A1 (es) | 2006-03-02 | 2008-04-23 | Athenix Corp | Metodos y composiciones para mejorar la actividad enzimatica en plantas transgenicas |
MX2008012252A (es) | 2006-03-24 | 2009-01-14 | Basf Plant Science Gmbh | Proteinas que se relacionan con la respuesta al estres abiotico y homologos. |
AR060523A1 (es) | 2006-04-19 | 2008-06-25 | Pioneer Hi Bred Int | Moleculas de polinucleotidos aislados que corresponden a alelos mutantes y tipo salvaje del gen de maiz d9 y metodos para usarlas |
EP2027276B1 (en) | 2006-05-16 | 2011-02-02 | Pioneer Hi-Bred International, Inc. | Antifungal polypeptides |
EP2018435B1 (en) | 2006-05-17 | 2012-07-11 | Pioneer Hi-Bred International Inc. | Artificial plant minichromosomes |
WO2007147096A2 (en) | 2006-06-15 | 2007-12-21 | Athenix Corporation | A family of pesticidal proteins and methods for their use |
US7994390B2 (en) | 2006-09-29 | 2011-08-09 | The Chinese University Of Hong Kong | Use of GmRD22-like genes to protect against abiotic stress |
US20080104726A1 (en) * | 2006-10-05 | 2008-05-01 | Ceres, Inc. | Fruit regulatory regions |
US20100162433A1 (en) | 2006-10-27 | 2010-06-24 | Mclaren James | Plants with improved nitrogen utilization and stress tolerance |
EP2099916A2 (en) * | 2006-12-22 | 2009-09-16 | Dow AgroSciences LLC | Plant-made west nile virus (wnv) vaccines, vectors and plant codon optimized sequences |
US8153863B2 (en) | 2007-03-23 | 2012-04-10 | New York University | Transgenic plants expressing GLK1 and CCA1 having increased nitrogen assimilation capacity |
US8329991B2 (en) | 2007-03-23 | 2012-12-11 | Basf Plant Science Gmbh | Transgenic plant with increased stress tolerance and yield |
EP2543731B1 (en) | 2007-04-04 | 2018-08-22 | Basf Se | Herbicide-resistant brassica plants and methods of use |
CA2682349C (en) | 2007-04-04 | 2017-08-22 | Basf Plant Science Gmbh | Ahas mutants |
WO2008145629A2 (en) | 2007-05-25 | 2008-12-04 | Cropdesign N.V. | Yield enhancement in plants by modulation of maize alfins |
CA2687320A1 (en) | 2007-05-29 | 2008-12-04 | Basf Plant Science Gmbh | Transgenic plants with increased stress tolerance and yield |
EP2179043A2 (en) | 2007-07-13 | 2010-04-28 | BASF Plant Science GmbH | Transgenic plants with increased stress tolerance and yield |
AU2008281715A1 (en) | 2007-08-02 | 2009-02-05 | Basf Plant Science Gmbh | Transgenic plants with increased stress tolerance and yield |
BR122018000387B1 (pt) | 2007-10-10 | 2019-05-21 | Athenix Corp. | Molécula de ácido nucleico isolada, vetor, célula hospedeira bacteriana, e método para proteção de uma planta contra uma praga |
US20090144852A1 (en) | 2007-10-16 | 2009-06-04 | Athenix Corporation | Axmi-066 and axmi-076: delta-endotoxin proteins and methods for their use |
CA2706799A1 (en) | 2007-11-27 | 2009-06-04 | Basf Plant Science Gmbh | Transgenic plants with increased stress tolerance and yield |
SG191671A1 (en) * | 2007-12-13 | 2013-07-31 | Danisco Us Inc | Cultured cells that produce isoprene and methods for producing isoprene |
BRPI0909285B1 (pt) * | 2008-03-31 | 2020-01-28 | Ceres Inc | ácido nucleico, construção de vetor, método de condução de transcrição, método de expressão de uma região de codificação exógena em uma planta, método de alteração da expressão de um gene em uma planta, método de produção de uma planta transgênica |
AU2009240505B2 (en) | 2008-04-23 | 2013-09-05 | Danisco Us Inc. | Isoprene synthase variants for improved microbial production of isoprene |
CA3008307C (en) | 2008-06-25 | 2020-12-15 | Athenix Corp. | Toxin genes and methods for their use |
EP2949659A1 (en) | 2008-07-02 | 2015-12-02 | Athenix Corporation | AXMI-115, AXMI-113, AXMI-005, AXMI-163 and AXMI-184: insecticidal proteins and methods for their use |
SI2331694T1 (sl) | 2008-07-31 | 2016-09-30 | Anglo Netherlands Grain Bv | Sončnične rastline odporne na herbicide |
EP2695944A3 (en) | 2008-08-20 | 2014-04-23 | BASF Plant Science GmbH | Transgenic plants with increased yield |
US8426204B2 (en) | 2008-09-08 | 2013-04-23 | Athenix Corporation | Chlamydomonas EPSPS chloroplast transit peptide (CTP) and expression cassettes and transgenic plants utilizing the CTP |
US20110302673A1 (en) | 2008-09-23 | 2011-12-08 | Basf Plant Science Gmbh | Transgenic Plants with Increased Yield |
MX2011003266A (es) | 2008-09-26 | 2011-07-20 | Basf Agrochemical Products Bv | Mutantes de acetohidroxiacido sintasa resistentes a herbicidas y metodos de uso. |
MX2011005604A (es) | 2008-11-28 | 2011-09-01 | Merial Ltd | Vacuna recombinante para influenza aviar y sus usos. |
US8431775B2 (en) | 2008-12-04 | 2013-04-30 | Pioneer Hi Bred International Inc | Methods and compositions for enhanced yield by targeted expression of knotted1 |
BRPI0923334A2 (pt) | 2008-12-15 | 2021-09-08 | Athenix Corporation | Gene que codifica toxinas de nematoide. |
US8293533B2 (en) | 2008-12-19 | 2012-10-23 | E.I. Du Pont De Nemours And Company | Site-specific integration and stacking of transgenes in soybean via DNA recombinase mediated cassette exchange |
MX2011006693A (es) | 2008-12-22 | 2011-10-14 | Athenix Corp | Genes pesticidas de brevibacillus y metodos para su uso. |
CN102388143A (zh) | 2008-12-23 | 2012-03-21 | 阿森尼克斯公司 | AXMI-150 δ-内毒素基因及其使用方法 |
CN101768213B (zh) | 2008-12-30 | 2012-05-30 | 中国科学院遗传与发育生物学研究所 | 一种与植物分蘖数目相关的蛋白及其编码基因与应用 |
EP2206723A1 (en) | 2009-01-12 | 2010-07-14 | Bonas, Ulla | Modular DNA-binding domains |
US20110239315A1 (en) | 2009-01-12 | 2011-09-29 | Ulla Bonas | Modular dna-binding domains and methods of use |
US20120042418A1 (en) | 2009-01-28 | 2012-02-16 | Basf Plant Science Company Gmbh | Engineering NF-YB Transcription Factors for Enhanced Drought Resistance and Increased Yield in Transgenic Plants |
CA2750456A1 (en) | 2009-01-28 | 2010-08-05 | Basf Plant Science Company Gmbh | Transgenic plants having altered nitrogen metabolism |
WO2010091230A1 (en) | 2009-02-05 | 2010-08-12 | Athenix Corporation | Variant axmi-r1 delta-endotoxin genes and methods for their use |
AR075240A1 (es) | 2009-02-06 | 2011-03-16 | Syngenta Participations Ag | Modificacion de enzima multidominio para la expresion en plantas |
US20120058523A1 (en) | 2009-02-17 | 2012-03-08 | Edenspace Systems Corporation | Tempering of cellulosic biomass |
MY189620A (en) * | 2009-02-20 | 2022-02-21 | Malaysian Palm Oil Board | A constitutive promoter from oil palm |
CN101817879A (zh) | 2009-02-26 | 2010-09-01 | 中国科学院遗传与发育生物学研究所 | 金属硫蛋白及其编码基因与应用 |
US8318900B2 (en) | 2009-02-27 | 2012-11-27 | Athenix Corp. | Pesticidal proteins and methods for their use |
EP2666866A3 (en) | 2009-03-06 | 2014-03-05 | Athenix Corporation | Methods and compositions for controlling plant pests |
CN102421792B (zh) | 2009-03-11 | 2015-11-25 | 阿森尼克斯公司 | Axmi-001、axmi-002、axmi-030、axmi-035和axmi-045: 来自苏云金芽孢杆菌的杀虫蛋白及其用法 |
BRPI1006259A2 (pt) | 2009-03-23 | 2015-08-25 | Basf Plant Science Co Gmbh | ''planta transgência transformada com um cassete de expressão, semente que é uma reprodução verdadeira para um transgene e método para aumentar a produção de uma planta'' |
CA2759700A1 (en) | 2009-04-23 | 2010-10-28 | Danisco Us Inc. | Three-dimensional structure of isoprene synthase and its use thereof for generating variants |
CN102439032B (zh) | 2009-05-04 | 2015-07-22 | 先锋国际良种公司 | 通过调节ap2转录因子增加植物中的产量 |
MY186558A (en) * | 2009-05-13 | 2021-07-27 | Malaysian Palm Oil Board | A constitutive promoter from oil palm type 2 |
US8471100B2 (en) * | 2009-05-15 | 2013-06-25 | University Of Tennessee Research Foundation | Environmental stress-inducible promoter and its application in crops |
US20120220015A1 (en) | 2009-05-26 | 2012-08-30 | Biolex Therapeutics | Compositions and methods for production of aglycosylated plasminogen |
WO2010138971A1 (en) | 2009-05-29 | 2010-12-02 | Edenspace Systems Corporation | Plant gene regulatory elements |
WO2010147825A1 (en) | 2009-06-09 | 2010-12-23 | Pioneer Hi-Bred International, Inc. | Early endosperm promoter and methods of use |
AU2010259295B2 (en) | 2009-06-10 | 2015-05-07 | Temasek Life Sciences Laboratory Limited | Virus induced gene silencing (VIGS) for functional analysis of genes in cotton. |
ES2609332T3 (es) | 2009-07-02 | 2017-04-19 | Athenix Corporation | Gen pesticida AXMI-205 y métodos para su uso |
MX354219B (es) | 2009-07-31 | 2018-02-19 | Athenix Corp | Familia de genes plaguicidas axmi-192 y metodos para su uso. |
MX2012004819A (es) | 2009-10-26 | 2012-06-25 | Pioneer Hi Bred Int | Promotor especifico de ovulo somatico y metodos de uso. |
CN101818157B (zh) * | 2009-12-02 | 2013-01-02 | 安徽省农业科学院水稻研究所 | 一种人工设计的Bt抗虫基因及其应用 |
CA2785653C (en) | 2009-12-28 | 2018-05-01 | Merial Limited | Recombinant ndv antigen and uses thereof |
US8704041B2 (en) | 2009-12-30 | 2014-04-22 | Pioneer Hi Bred International Inc | Methods and compositions for targeted polynucleotide modification |
EP2529018B1 (en) | 2009-12-30 | 2016-06-22 | Pioneer Hi-Bred International, Inc. | Methods and compositions for the introduction and regulated expression of genes in plants |
BR112012018618A2 (pt) | 2010-01-26 | 2017-01-10 | Pioneer Hi Bred Int | método para seleção de uma planta ou germoplasma apresentando tolerãncia, tolerãncia aumentada, susceptibilidade, ou susceptibilidade aumentada a um ou mais herbicidas, planta, kit para seleção de pelo menos uma planta de soja, método de triagem de uma soja, método para controlar de modo seletivo plantas daninhas, método para triar de modo seletivo plantas de soja para tolerância a um ou mais herbicidas |
CN102892886A (zh) | 2010-02-18 | 2013-01-23 | 阿森尼克斯公司 | AXMI218、AXMI219、AXMI220、AXMI226、AXMI227、AXMI228、AXMI229、AXMI230、和AXMI231δ-内毒素基因以及它们的使用方法 |
EP2536267B1 (en) | 2010-02-18 | 2015-07-22 | Athenix Corp. | AXMI221z, AXMI222z, AXMI223z, AXMI224z, AND AXMI225z DELTA-ENDOTOXIN GENES AND METHODS FOR THEIR USE |
US20110236416A1 (en) | 2010-03-12 | 2011-09-29 | Jean-Christophe Audonnet | Foot and mouth disease virus recombinant vaccines and uses thereof |
BR112012028132A2 (pt) | 2010-05-04 | 2015-09-15 | Basf Se | plantas que tem tolerância aumentada a herbicidas |
WO2011139431A1 (en) | 2010-05-06 | 2011-11-10 | Pioneer Hi-Bred International, Inc. | Maize acc synthase 3 gene and protein and uses thereof |
US9234208B1 (en) * | 2010-05-10 | 2016-01-12 | Dow Agrosciences Llc | DIG-13 insecticidal cry toxins |
FR2961375B1 (fr) | 2010-06-16 | 2016-05-13 | Inst De Rech Pour Le Dev (Ird) | Surproduction d'acide jasmonique dans des plantes transgeniques |
UA112969C2 (uk) | 2010-08-03 | 2016-11-25 | Сібас Юс Ллс | Рослина, стійка до одного або більше ррх-інгібуючих гербіцидів, яка містить мутантний ген протопорфіриноген ix оксидази (ррх) |
US20120122223A1 (en) | 2010-08-03 | 2012-05-17 | Cibus Us Llc | Mutated protoporphyrinogen ix oxidase (ppx) genes |
US8993837B2 (en) | 2010-08-13 | 2015-03-31 | Pioneer Hi-Bred International, Inc | Chimeric promoters and methods of use |
CA2807785A1 (en) | 2010-08-23 | 2012-03-01 | Pioneer Hi-Bred International, Inc. | Novel defensin variants and methods of use |
EP2633043A2 (en) | 2010-10-27 | 2013-09-04 | Danisco US Inc. | Isoprene synthase variants for improved production of isoprene |
US9018451B2 (en) | 2010-12-03 | 2015-04-28 | M S Technologies, LLC | Optimized expression of glyphosate resistance encoding nucleic acid molecules in plant cells |
US11274313B2 (en) | 2010-12-16 | 2022-03-15 | BASF Agro B.V. | Plants having increased tolerance to herbicides |
BR112013014698A2 (pt) | 2010-12-22 | 2017-03-07 | E I Du Point De Nemours & Company | constructo de dna, vetor, célula vegetal, planta, semente transgênica proveniente da planta, método para expressão de uma sequência de nucleotídeos em uma planta, método para expressão de uma sequência de nucleotídeos em uma célula vegetal, método para expressar seletivamente uma sequência de nucleotídeos em tecidos de raiz, caule, grão e pendão de milho |
US8895716B2 (en) | 2010-12-22 | 2014-11-25 | Pioneer Hi-Bred International, Inc. | Viral promoter, truncations thereof, and methods of use |
BR112013015515A2 (pt) | 2010-12-28 | 2018-04-24 | Pioneer Hi Bred Int | molécula de ácido nucleico isolada, construto de dna, célula hospedeira, planta transgênica, semente transformada da planta, polipeptídeo isolado com atividade pesticida, composição, método para controlar uma população de praga de lepidóptero, método para matar uma praga de lepidóptero, método para produzir um polipeptídeo com atividade pesticida, planta que tem incorporado de maneira estável em seu genoma um construto de dna, método para proteger uma planta contra uma praga |
US20140137292A1 (en) | 2011-01-14 | 2014-05-15 | University Of Florida Research Foundation Inc. | Citrus trees with resistance to citrus canker |
CN103796507B (zh) | 2011-02-01 | 2017-09-01 | 科罗拉多小麦研究基金会公司 | 乙酰辅酶a羧化酶除草剂抗性植物 |
CN103459601A (zh) | 2011-02-11 | 2013-12-18 | 先锋国际良种公司 | 具有对抗玉米根虫的活性的合成杀昆虫蛋白 |
EP2675900B1 (en) | 2011-02-15 | 2017-09-06 | Pioneer Hi-Bred International, Inc. | Root-preferred promoter and methods of use |
US8878007B2 (en) | 2011-03-10 | 2014-11-04 | Pioneer Hi Bred International Inc | Bacillus thuringiensis gene with lepidopteran activity |
US8648230B2 (en) | 2011-03-18 | 2014-02-11 | Ms Technologies, Llc | Regulatory regions preferentially expressing in non-pollen plant tissue |
US10030245B2 (en) | 2011-03-23 | 2018-07-24 | E I Du Pont De Nemours And Company | Methods for producing a complex transgenic trait locus |
US9695440B2 (en) | 2011-03-30 | 2017-07-04 | Athenix Corp. | AXMI232, AXMI233, and AXMI249 toxin genes and methods for their use |
US9090906B2 (en) | 2011-03-30 | 2015-07-28 | Universidad Nacional Autonoma De Mexico | Mutant Bacillus thuringiensis cry genes and methods of use |
US9321814B2 (en) | 2011-03-30 | 2016-04-26 | Athenix Corp. | AXMI238 toxin gene and methods for its use |
AR085939A1 (es) | 2011-04-05 | 2013-11-06 | Athenix Corp | Gen insecticida variante axmi115 y metodos para utilizarlo |
US9150625B2 (en) | 2011-05-23 | 2015-10-06 | E I Du Pont De Nemours And Company | Chloroplast transit peptides and methods of their use |
EP2721160A2 (en) | 2011-06-14 | 2014-04-23 | Synthon Biopharmaceuticals B.V. | Compositions and methods for making and biocontaining auxotrophic transgenic plants |
US20130097734A1 (en) | 2011-07-12 | 2013-04-18 | Two Blades Foundation | Late blight resistance genes |
RU2014107672A (ru) | 2011-07-28 | 2015-09-10 | Атеникс Корп. | Ген токсина axmi270 и способы его применения |
PL2737069T3 (pl) | 2011-07-28 | 2018-01-31 | Athenix Corp | Warianty białka axmi205 i sposoby ich zastosowania |
CA2844355C (en) | 2011-07-29 | 2022-05-10 | Athenix Corp. | Pesticidal gene with pesticidal activity against western corn rootworm and method for its use |
WO2013019411A1 (en) | 2011-08-03 | 2013-02-07 | E. I. Du Pont De Nemours And Company | Methods and compositions for targeted integration in a plant |
US11377663B1 (en) | 2011-08-30 | 2022-07-05 | Monsanto Technology Llc | Genetic regulatory elements |
AU2012301912A1 (en) | 2011-08-31 | 2014-03-06 | E. I. Dupont De Nemours & Company | Methods for tissue culture and transformation of sugarcane |
CN104053770A (zh) | 2011-10-19 | 2014-09-17 | 凯金公司 | 用于产生肉桂酸内酯和/或补身二醇的方法 |
WO2013063344A1 (en) | 2011-10-28 | 2013-05-02 | Pioneer Hi-Bred International, Inc. | Engineered pep carboxylase variants for improved plant productivity |
FR2984076A1 (fr) | 2011-12-15 | 2013-06-21 | Inst Rech Developpement Ird | Surproduction de jasmonates dans des plantes transgeniques |
CA2860611A1 (en) | 2012-01-06 | 2013-07-11 | Pioneer Hi-Bred International, Inc. | Compositions and methods for the expression of a sequence in a reproductive tissue of a plant |
EP2800816A1 (en) | 2012-01-06 | 2014-11-12 | Pioneer Hi-Bred International Inc. | Ovule specific promoter and methods of use |
US9006515B2 (en) | 2012-01-06 | 2015-04-14 | Pioneer Hi Bred International Inc | Pollen preferred promoters and methods of use |
MX2014008243A (es) | 2012-01-06 | 2015-02-20 | Pioner Hi Bred International Inc | Un metodo para seleccionar plantas por elementos geneticos que inducen la partenogenesis en plantas. |
BR112014018294B1 (pt) | 2012-01-26 | 2022-01-11 | Norfolk Plant Sciences, Ltd | Método para produzir uma planta, cassete de expressão, e, célula bacteriana |
AR089793A1 (es) | 2012-01-27 | 2014-09-17 | Du Pont | Metodos y composiciones para generar locus de rasgos transgenicos complejos |
CA2866239C (en) | 2012-03-08 | 2020-07-28 | Athenix Corp. | Bacillus thuringiensis toxin gene axmi335 and methods for its use |
UY34663A (es) | 2012-03-08 | 2013-10-31 | Athenix Corp | Acido nucleico y proteina recombinante del gen de la endotoxina axmi345, vectores, células, composiciones y métodos para proteger plantas |
WO2013134651A1 (en) | 2012-03-09 | 2013-09-12 | Board Of Trustees Of Michigan State University | Method of enhancing plant drought tolerance by expression of ndr1 |
US9163263B2 (en) | 2012-05-02 | 2015-10-20 | The Goodyear Tire & Rubber Company | Identification of isoprene synthase variants with improved properties for the production of isoprene |
BR112014027468A2 (pt) | 2012-05-04 | 2017-06-27 | Du Pont | polinucleotídeo isolado ou recombinante, construção de dna recombinante, célula, planta, explante vegetal, semente transgênica, polipeptídeo isolado, composição, métodos de produção de meganuclease, de introdução de rompimento e de integração de um polinucleotídeo. |
US20150119325A1 (en) | 2012-05-29 | 2015-04-30 | North Carolina Central University | Methods for the production of cytoprotective asialo-erythropoietin in plants and its purification from plant tissues |
CN104540951B (zh) | 2012-06-07 | 2017-07-04 | 美国陶氏益农公司 | 使用芸苔属双向组成型启动子表达转基因的构建体和方法 |
US9347105B2 (en) | 2012-06-15 | 2016-05-24 | Pioneer Hi Bred International Inc | Genetic loci associated with resistance of soybean to cyst nematode and methods of use |
BR112014031260A2 (pt) | 2012-06-15 | 2019-08-20 | Du Pont | métodos e composições que envolvem variantes de als com preferência de substrato nativo |
AR091489A1 (es) | 2012-06-19 | 2015-02-11 | Basf Se | Plantas que tienen una mayor tolerancia a herbicidas inhibidores de la protoporfirinogeno oxidasa (ppo) |
US10041087B2 (en) | 2012-06-19 | 2018-08-07 | BASF Agro B.V. | Plants having increased tolerance to herbicides |
CN103998610B (zh) | 2012-06-29 | 2020-03-17 | 阿森尼克斯公司 | Axmi277线虫毒素及其使用方法 |
CA2882143A1 (en) | 2012-07-26 | 2014-01-30 | Dow Agrosciences Llc | High-throughput dna fragment assembly |
JP2015524672A (ja) | 2012-08-17 | 2015-08-27 | ダウ アグロサイエンシィズ エルエルシー | 植物における導入遺伝子発現のためのトウモロコシ非翻訳領域の使用 |
US9758793B2 (en) | 2012-08-30 | 2017-09-12 | Athenix Corp. | AXMI-234 and AXMI-235 delta-endotoxin genes and methods for their use |
WO2014042923A1 (en) | 2012-09-13 | 2014-03-20 | Indiana University Research & Technology Corporation | Compositions and systems for conferring disease resistance in plants and methods of use thereof |
US10793872B2 (en) | 2012-09-14 | 2020-10-06 | BASF Agricultural Solutions Seed US LLC | HPPD variants and methods of use |
CA2887571A1 (en) | 2012-10-11 | 2014-04-17 | Pioneer Hi-Bred International, Inc. | Guard cell promoters and uses thereof |
US9783820B2 (en) | 2012-10-15 | 2017-10-10 | Pioneer Hi-Bred International, Inc. | Methods and compositions to enhance activity of Cry endotoxins |
CN102994514B (zh) * | 2012-11-07 | 2015-06-10 | 武汉禾元生物科技股份有限公司 | 一种从水稻种子生产和分离纯化重组人抗胰蛋白酶(OsrAAT)的方法 |
EP2920319B1 (en) | 2012-11-16 | 2020-02-19 | Poseida Therapeutics, Inc. | Site-specific enzymes and methods of use |
US20140173775A1 (en) | 2012-12-13 | 2014-06-19 | Pioneer Hi-Bred International, Inc. | Methods and compositions for producing and selecting transgenic plants |
US10253325B2 (en) | 2012-12-19 | 2019-04-09 | Boston Medical Center Corporation | Methods for elevating fat/oil content in plants |
WO2014100525A2 (en) | 2012-12-21 | 2014-06-26 | Pioneer Hi-Bred International, Inc. | Compositions and methods for auxin-analog conjugation |
US20140203176A1 (en) | 2013-01-23 | 2014-07-24 | Dow Agrosciences Llc | Systems and methods for real-time sampling and analysis of biomolecules beneath the surface of biological tissue |
CA2901160C (en) | 2013-02-13 | 2023-08-22 | Athenix Corp. | Use of axmi184 for the control of rootworm insects |
US9783817B2 (en) | 2013-03-04 | 2017-10-10 | Arkansas State University | Methods of expressing and detecting activity of expansin in plant cells |
AU2014225732B2 (en) | 2013-03-07 | 2020-03-19 | BASF Agricultural Solutions Seed US LLC | Toxin genes and methods for their use |
CA2905399A1 (en) | 2013-03-11 | 2014-10-09 | Pioneer Hi-Bred International, Inc. | Methods and compositions employing a sulfonylurea-dependent stabilization domain |
WO2014164775A1 (en) | 2013-03-11 | 2014-10-09 | Pioneer Hi-Bred International, Inc. | Methods and compositions to improve the spread of chemical signals in plants |
US9803214B2 (en) | 2013-03-12 | 2017-10-31 | Pioneer Hi-Bred International, Inc. | Breeding pair of wheat plants comprising an MS45 promoter inverted repeat that confers male sterility and a construct that restores fertility |
RU2694686C2 (ru) | 2013-03-12 | 2019-07-16 | Е.И.Дюпон Де Немур Энд Компани | Способы идентификации вариантных сайтов распознавания для редкощепящих сконструированных средств для индукции двунитевого разрыва, и композиции с ними, и их применения |
US9273322B2 (en) | 2013-03-12 | 2016-03-01 | Pioneer Hi Bred International Inc | Root-preferred promoter and methods of use |
US9243258B2 (en) | 2013-03-12 | 2016-01-26 | Pioneer Hi Bred International Inc | Root-preferred promoter and methods of use |
US9416368B2 (en) | 2013-03-13 | 2016-08-16 | E I Du Pont De Nemours And Company | Identification of P. pachyrhizi protein effectors and their use in producing Asian soybean rust (ASR) resistant plants |
US20140289906A1 (en) | 2013-03-14 | 2014-09-25 | Pioneer Hi-Bred International, Inc. | Compositions Having Dicamba Decarboxylase Activity and Methods of Use |
CA2905128C (en) | 2013-03-14 | 2023-01-03 | Cibus Us Llc | Mutated allene oxide synthase 2 (aos2) genes |
BR112015023286A2 (pt) | 2013-03-14 | 2018-03-06 | Arzeda Corp | polipeptídeo recombinante com atividade da dicamba descarboxilase, construto de polinucleotídeo, célula, método de produção de uma célula hospedeira compreendendo um polinucleotídeo heterólogo que codifica um polipeptídeo tendo atividade da dicamba descarboxilase, método para descarboxilar dicamba, um derivado de dicamba ou um metabolito de dicamba, método para a detecção de um polipeptideo e método para a detecção da presença de um polinucleotideo que codifica um polipeptideo tendo atividade da dicamba descarboxilase |
US9957515B2 (en) | 2013-03-15 | 2018-05-01 | Cibus Us Llc | Methods and compositions for targeted gene modification |
KR102339732B1 (ko) | 2013-03-15 | 2021-12-15 | 시버스 유에스 엘엘씨 | 올리고뉴클레오타이드 매개 유전자 보수를 사용한 표적화된 유전자 변형의 효율을 증가시키기 위한 방법 및 조성물 |
CA2901316A1 (en) | 2013-03-15 | 2014-09-25 | Pioneer Hi-Bred International, Inc. | Phi-4 polypeptides and methods for their use |
EP2970405B1 (en) | 2013-03-15 | 2018-09-12 | Bayer Cropscience LP | Constitutive soybean promoters |
WO2014204988A2 (en) | 2013-06-17 | 2014-12-24 | Asana Biosciences, Llc | 5t4-targeted immunofusion molecule and methods |
WO2015006105A1 (en) | 2013-07-09 | 2015-01-15 | Board Of Trustees Of Michigan State University | Transgenic plants produced with a k-domain, and methods and expression cassettes related thereto |
US11459579B2 (en) | 2013-07-09 | 2022-10-04 | Board Of Trustees Of Michigan State University | Transgenic plants produced with a K-domain, and methods and expression cassettes related thereto |
EP3030072B1 (en) | 2013-08-08 | 2020-03-04 | Pioneer Hi-Bred International, Inc. | Insecticidal polypeptides having broad spectrum activity and uses thereof |
US9862967B2 (en) | 2013-08-09 | 2018-01-09 | Athenix Corp. | AXMI281 toxin gene and methods for its use |
UY35696A (es) | 2013-08-09 | 2015-03-27 | Athenix Corp | ?molécula de adn recombinante que comprende al gen de toxina axmi440, vector, célula huésped, plantas, composiciones y métodos relacionados?. |
US10968462B2 (en) | 2013-08-12 | 2021-04-06 | BASF Agro B.V. | Plants having increased tolerance to herbicides |
WO2015022636A2 (en) | 2013-08-12 | 2015-02-19 | Basf Se | Plants having increased tolerance to herbicides |
WO2015023846A2 (en) | 2013-08-16 | 2015-02-19 | Pioneer Hi-Bred International, Inc. | Insecticidal proteins and methods for their use |
WO2015026887A1 (en) | 2013-08-22 | 2015-02-26 | E. I. Du Pont De Nemours And Company | A soybean u6 polymerase iii promoter and methods of use |
US20160201073A1 (en) | 2013-09-11 | 2016-07-14 | Pioneer Hi-Bred International, Inc. | Plant regulatory elements and methods of use thereof |
ES2937045T3 (es) | 2013-09-13 | 2023-03-23 | Pioneer Hi Bred Int | Proteínas insecticidas y métodos para su uso |
CN105764343B (zh) | 2013-11-25 | 2020-04-17 | 拜耳作物科学有限合伙公司 | 使用axmi-011控制半翅目昆虫 |
CN111560385B (zh) | 2013-12-09 | 2023-11-28 | 巴斯夫农业解决方案种子美国有限责任公司 | Axmi477、axmi482、axmi486和axmi525毒素基因及使用方法 |
WO2015116680A1 (en) | 2014-01-30 | 2015-08-06 | Two Blades Foundation | Plants with enhanced resistance to phytophthora |
WO2015120270A1 (en) | 2014-02-07 | 2015-08-13 | Pioneer Hi Bred International, Inc. | Insecticidal proteins and methods for their use |
MX2016010187A (es) | 2014-02-07 | 2017-07-11 | Pioneer Hi Bred Int | Proteinas insecticidas y metodos para su uso. |
MX2016011745A (es) | 2014-03-11 | 2017-09-01 | Bayer Cropscience Lp | Variantes de hppd y metodos de uso. |
CA2942407C (en) | 2014-03-14 | 2023-09-26 | Cibus Us Llc | Methods and compositions for increasing efficiency of targeted gene modification using oligonucleotide-mediated gene repair |
WO2015150465A2 (en) | 2014-04-03 | 2015-10-08 | Basf Se | Plants having increased tolerance to herbicides |
WO2015171603A1 (en) | 2014-05-06 | 2015-11-12 | Two Blades Foundation | Methods for producing plants with enhanced resistance to oomycete pathogens |
US11584936B2 (en) | 2014-06-12 | 2023-02-21 | King Abdullah University Of Science And Technology | Targeted viral-mediated plant genome editing using CRISPR /Cas9 |
AR100874A1 (es) | 2014-06-16 | 2016-11-09 | Consejo Nac De Investig Científicas Y Técnicas (Conicet) | Genes quiméricos y proteínas de resistencia oxidativa, y plantas transgénicas que incluyen los mismos |
US10676754B2 (en) | 2014-07-11 | 2020-06-09 | E I Du Pont De Nemours And Company | Compositions and methods for producing plants resistant to glyphosate herbicide |
US20170218384A1 (en) | 2014-08-08 | 2017-08-03 | Pioneer Hi-Bred International, Inc. | Ubiquitin promoters and introns and methods of use |
CN108064129A (zh) | 2014-09-12 | 2018-05-22 | 纳幕尔杜邦公司 | 玉米和大豆中复合性状基因座的位点特异性整合位点的产生和使用方法 |
BR102014023930A2 (pt) | 2014-09-26 | 2016-05-17 | Ctc Ct De Tecnologia Canavieira S A | método para transformar uma célula de planta ou tecido de planta usando agrobacterium, planta transgênica, célula transgênica ou tecido transgênico, meio de cultura, e, uso de um método para transformar uma célula de planta ou tecido de planta |
BR112017007932A2 (pt) | 2014-10-16 | 2018-01-23 | Du Pont | proteínas inseticidas e métodos para uso das mesmas |
EP3207126A4 (en) | 2014-10-16 | 2018-04-04 | Pioneer Hi-Bred International, Inc. | Insecticidal polypeptides having broad spectrum activity and uses thereof |
CN107074917B (zh) | 2014-10-16 | 2022-05-24 | 先锋国际良种公司 | 具有改进的活性谱的杀昆虫多肽及其用途 |
CA2971425A1 (en) | 2014-12-16 | 2016-06-23 | Pioneer Hi-Bred International, Inc. | Restoration of male fertility in wheat |
WO2016106184A1 (en) | 2014-12-22 | 2016-06-30 | AgBiome, Inc. | Methods for making a synthetic gene |
CA2972881A1 (en) | 2015-01-21 | 2016-07-28 | Basf Se | Plants having increased tolerance to herbicides |
CN107406858A (zh) | 2015-02-25 | 2017-11-28 | 先锋国际良种公司 | 用于指导rna/cas内切核酸酶复合物的调节型表达的组合物和方法 |
CA2977026A1 (en) | 2015-03-11 | 2016-09-15 | E.I. Du Pont De Nemours And Company | Insecticidal combinations of pip-72 and methods of use |
EP3270685B1 (en) | 2015-03-19 | 2023-07-19 | Pioneer Hi-Bred International, Inc. | Methods and compositions for accelerated trait introgression |
US20160304898A1 (en) | 2015-04-17 | 2016-10-20 | AgBiome, Inc. | Pesticidal Genes and Methods of Use |
BR112017022852A2 (pt) | 2015-04-22 | 2018-07-17 | Agbiome Inc | polipeptídeo recombinante com atividade pesticida, composição, molécula de ácido nucleico recombinante, ácido nucleico recombinante, célula hospedeira, construto de dna, vetor, método para controlar uma população de pragas, método para produzir um polipeptídeo, planta, semente transgênica, método para proteger uma planta e método para aumentar o rendimento em uma planta |
WO2016182881A1 (en) | 2015-05-09 | 2016-11-17 | Two Blades Foundation | Late blight resistance gene from solanum americanum and methods of use |
WO2016186946A1 (en) | 2015-05-15 | 2016-11-24 | Pioneer Hi-Bred International, Inc. | Rapid characterization of cas endonuclease systems, pam sequences and guide rna elements |
CN116333064A (zh) | 2015-05-19 | 2023-06-27 | 先锋国际良种公司 | 杀昆虫蛋白及其使用方法 |
MA46313A (fr) | 2015-06-03 | 2021-04-28 | Agbiome Inc | Gènes pesticides et leurs procédés d'utilisation |
EP3929286A1 (en) | 2015-06-17 | 2021-12-29 | Poseida Therapeutics, Inc. | Compositions and methods for directing proteins to specific loci in the genome |
MX2017017146A (es) | 2015-06-22 | 2018-02-23 | Agbiome Inc | Genes pesticidas y metodos de uso. |
EP3943602A1 (en) | 2015-08-06 | 2022-01-26 | Pioneer Hi-Bred International, Inc. | Plant derived insecticidal proteins and methods for their use |
WO2017062790A1 (en) | 2015-10-09 | 2017-04-13 | Two Blades Foundation | Cold shock protein receptors and methods of use |
MX2018004808A (es) | 2015-10-20 | 2018-08-01 | Pioneer Hi Bred Int | Funcion restauradora para un producto genico no funcional mediante sistemas cas guiados y metodos de uso. |
AR106931A1 (es) | 2015-10-22 | 2018-03-07 | Basf Se | Plantas que tienen una mayor tolerancia a herbicidas |
BR112018007796A2 (pt) | 2015-11-06 | 2018-10-30 | Du Pont | plantas de soja, partes de plantas de soja ou sementes de soja, método para selecionar uma célula de soja, métodos de seleção de uma célula de soja e de produção de um locus e molécula de ácido nucleico |
WO2017083092A1 (en) | 2015-11-10 | 2017-05-18 | Dow Agrosciences Llc | Methods and systems for predicting the risk of transgene silencing |
CN115786368A (zh) | 2015-11-30 | 2023-03-14 | 先锋国际良种公司 | 植物调节元件及其使用方法 |
EP3387134B1 (en) | 2015-12-11 | 2020-10-14 | Danisco US Inc. | Methods and compositions for enhanced nuclease-mediated genome modification and reduced off-target site effects |
US20180325119A1 (en) | 2015-12-18 | 2018-11-15 | Pioneer Hi-Bred International, Inc. | Insecticidal proteins and methods for their use |
DK3390631T3 (da) | 2015-12-18 | 2020-07-13 | Danisco Us Inc | Fremgangsmåder og sammensætninger til t-rna-baseret guide-rna-ekspression |
WO2017112538A2 (en) | 2015-12-22 | 2017-06-29 | AgBiome, Inc. | Pesticidal genes and methods of use |
US11104911B2 (en) | 2015-12-22 | 2021-08-31 | Pioneer Hi-Bred International, Inc. | Embryo-preferred Zea mays promoters and methods of use |
WO2017138986A1 (en) | 2016-02-09 | 2017-08-17 | Cibus Us Llc | Methods and compositions for increasing efficiency of targeted gene modification using oligonucleotide-mediated gene repair |
WO2017155715A1 (en) | 2016-03-11 | 2017-09-14 | Pioneer Hi-Bred International, Inc. | Novel cas9 systems and methods of use |
WO2017155714A1 (en) | 2016-03-11 | 2017-09-14 | Pioneer Hi-Bred International, Inc. | Novel cas9 systems and methods of use |
WO2017155717A1 (en) | 2016-03-11 | 2017-09-14 | Pioneer Hi-Bred International, Inc. | Novel cas9 systems and methods of use |
MX2018014245A (es) | 2016-05-20 | 2019-03-28 | Basf Agro Bv | Peptidos de transito dual para el direccionamiento a polipeptidos. |
EP3469077A1 (en) | 2016-06-14 | 2019-04-17 | Pioneer Hi-Bred International, Inc. | Use of cpf1 endonuclease for plant genome modifications |
WO2017222773A1 (en) | 2016-06-20 | 2017-12-28 | Pioneer Hi-Bred International, Inc. | Novel cas systems and methods of use |
CN109688807B (zh) | 2016-07-15 | 2023-04-28 | 巴斯夫欧洲公司 | 具有增加的除草剂耐受性的植物 |
CN109790153A (zh) | 2016-07-27 | 2019-05-21 | 巴斯夫农业公司 | 具有增加的除草剂耐受性的植物 |
MX2019002608A (es) | 2016-09-06 | 2019-06-06 | Agbiome Inc | Genes pesticidas y metodos de uso. |
CA3043493A1 (en) | 2016-11-23 | 2018-05-31 | BASF Agricultural Solutions Seed US LLC | Axmi669 and axmi991 toxin genes and methods for their use |
CN110462047A (zh) | 2016-12-16 | 2019-11-15 | 双刃基金会 | 晚疫病抗性基因及使用方法 |
CA3044728A1 (en) | 2016-12-20 | 2018-06-28 | BASF Agro B.V. | Plants having increased tolerance to herbicides |
JP2020504609A (ja) | 2016-12-22 | 2020-02-13 | ビーエーエスエフ アグリカルチュラル ソリューションズ シード ユーエス エルエルシー | 有害線虫の防除のためのcry14の使用 |
UY37570A (es) | 2017-01-18 | 2018-08-31 | Bayer Cropscience Lp | Uso de bp005 para el control de patógenos de planta |
CA3049775A1 (en) | 2017-01-18 | 2018-07-26 | BASF Agricultural Solutions Seed US LLC | Bp005 toxin gene and methods for its use |
US10793610B2 (en) | 2017-01-30 | 2020-10-06 | AgBiome, Inc. | Pesticidal genes and methods of use |
US11180770B2 (en) | 2017-03-07 | 2021-11-23 | BASF Agricultural Solutions Seed US LLC | HPPD variants and methods of use |
WO2018165091A1 (en) | 2017-03-07 | 2018-09-13 | Bayer Cropscience Lp | Hppd variants and methods of use |
US11371056B2 (en) | 2017-03-07 | 2022-06-28 | BASF Agricultural Solutions Seed US LLC | HPPD variants and methods of use |
BR112019021380A2 (pt) | 2017-04-11 | 2020-05-05 | Agbiome Inc | genes pesticidas e métodos de uso |
CN110891965A (zh) | 2017-04-24 | 2020-03-17 | 杜邦营养生物科学有限公司 | 植物中使用的抗crispr蛋白的方法和组合物 |
US20200407737A1 (en) | 2017-05-03 | 2020-12-31 | KWS SAAT SE & Co. KGaA | Use of crispr-cas endonucleases for plant genome engineering |
WO2018209209A1 (en) | 2017-05-12 | 2018-11-15 | Two Blades Foundation | Methods for screening proteins for pattern recognition receptor function in plant protoplasts |
EP3658675A1 (en) | 2017-07-28 | 2020-06-03 | Two Blades Foundation | Potyvirus resistance genes and methods of use |
US20190040411A1 (en) | 2017-08-03 | 2019-02-07 | AgBiome, Inc. | Pesticidal genes and methods of use |
CN111373046A (zh) | 2017-09-25 | 2020-07-03 | 先锋国际良种公司 | 组织偏好性启动子和使用方法 |
US20210032651A1 (en) | 2017-10-24 | 2021-02-04 | Basf Se | Improvement of herbicide tolerance to hppd inhibitors by down-regulation of putative 4-hydroxyphenylpyruvate reductases in soybean |
US11279944B2 (en) | 2017-10-24 | 2022-03-22 | BASF Agricultural Solutions Seed US LLC | Of herbicide tolerance to 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors by down-regulation of HPPD expression in soybean |
WO2019108619A1 (en) | 2017-11-28 | 2019-06-06 | Two Blades Foundation | Methods and compositions for enhancing the disease resistance of plants |
EP3716765A4 (en) | 2017-11-29 | 2021-08-25 | Basf Se | PLANTS WITH INCREASED TOLERANCE TO HERBICIDES |
JP7384801B2 (ja) | 2017-12-15 | 2023-11-21 | ダニスコ・ユーエス・インク | Cas9変異体及び使用方法 |
BR112020012477A2 (pt) | 2017-12-19 | 2020-11-24 | Pioneer Hi-Bred International, Inc. | polipeptídeo recombinante; polipeptídeo inseticida recombinante; composição agrícola; construto de dna; célula hospedeira; planta transgênica; método para inibir o crescimento ou exterminar uma praga de inseto ou população de praga; método para controlar a infestação de praga; e método para melhorar o rendimento de uma cultura |
WO2019126578A1 (en) | 2017-12-20 | 2019-06-27 | Poseida Therapeutics, Inc. | Compositions and methods for directing proteins to specific loci in the genome |
US10907173B2 (en) | 2017-12-22 | 2021-02-02 | AgBiome, Inc. | Pesticidal genes and methods of use |
WO2019133371A1 (en) | 2017-12-27 | 2019-07-04 | Pioneer Hi-Bred International, Inc. | Transformation of dicot plants |
CA3087266A1 (en) | 2018-01-09 | 2019-07-18 | Cibus Us Llc | Shatterproof genes and mutations |
US11732271B2 (en) | 2018-01-12 | 2023-08-22 | The Sainsbury Laboratory | Stem rust resistance genes and methods of use |
WO2019142099A1 (en) | 2018-01-17 | 2019-07-25 | Basf Se | Plants having increased tolerance to herbicides |
US11905518B2 (en) | 2018-02-12 | 2024-02-20 | Curators Of The University Of Missouri | Small auxin upregulated (SAUR) gene for the improvement of root system architecture, waterlogging tolerance, drought resistance and yield in plants and methods of uses |
WO2019204584A1 (en) | 2018-04-20 | 2019-10-24 | AgBiome, Inc. | Pesticidal proteins and methods of use |
CN108660151A (zh) * | 2018-05-25 | 2018-10-16 | 中国林业科学研究院亚热带林业研究所 | 一种毛竹遗传转化的方法 |
AU2019282149A1 (en) | 2018-06-05 | 2021-01-21 | LifeEDIT Therapeutics, Inc. | RNA-guided nucleases and active fragments and variants thereof and methods of use |
US20210277409A1 (en) | 2018-06-28 | 2021-09-09 | Pioneer Hi-Bred International, Inc. | Methods for selecting transformed plants |
US20210324398A1 (en) | 2018-06-29 | 2021-10-21 | Pioneer Hi-Bred International, Inc. | Edited nac genes in plants |
CA3111090A1 (en) | 2018-10-31 | 2020-05-07 | Pioneer Hi-Bred International, Inc. | Compositions and methods for ochrobactrum-mediated plant transformation |
EP4339286A2 (en) | 2018-12-27 | 2024-03-20 | LifeEDIT Therapeutics, Inc. | Polypeptides useful for gene editing and methods of use |
BR112021017998A2 (pt) | 2019-03-11 | 2021-11-16 | Pioneer Hi Bred Int | Métodos para a produção de plantas clonais |
CA3128376A1 (en) | 2019-03-27 | 2020-10-01 | Pioneer Hi-Bred International, Inc. | Plant explant transformation |
WO2020198496A1 (en) | 2019-03-28 | 2020-10-01 | Pioneer Hi-Bred International, Inc. | Modified agrobacterium strains and use thereof for plant transformation |
WO2020206197A1 (en) | 2019-04-05 | 2020-10-08 | Danisco Us Inc | Methods for polynucleotide integration into the genome of bacillus using dual circular recombinant dna constructs and compositions thereof |
JP2022526982A (ja) | 2019-04-05 | 2022-05-27 | ダニスコ・ユーエス・インク | 線状組換えDNAコンストラクトを使用してバチルス(Bacillus)のゲノムにドナーDNA配列を組み込むための方法及びその組成物 |
BR112021020770A2 (pt) | 2019-04-18 | 2022-01-04 | Pioneer Hi Bred Int | Fatores de embriogênese para reprogramação celular de uma célula vegetal |
BR112021026248A2 (pt) | 2019-06-27 | 2022-03-03 | Two Blades Found | Molécula de ácido nucleico isolada que codifica uma proteína atrlp23 engenheirada, proteína atrlp23 engenheirada, cassete de expressão, vetor, célula hospedeira, planta ou célula vegetal, métodos para fazer uma proteína atrlp23 engenheirada, para fazer uma molécula de ácido nucleico que codifica uma proteína atrlp23 engenheirada e para intensificar a resistência de uma planta |
WO2021011348A1 (en) | 2019-07-12 | 2021-01-21 | The Regents Of The University Of California | Plants with enhanced resistance to bacterial pathogens |
CA3147783A1 (en) | 2019-08-12 | 2021-02-18 | Tyson D. BOWEN | Rna-guided nucleases and active fragments and variants thereof and methods of use |
CN114981428A (zh) | 2019-11-19 | 2022-08-30 | 丹尼斯科美国公司 | 用于修饰芽孢杆菌属基因组的无选择标记方法及其组合物 |
JP2023508731A (ja) | 2019-12-30 | 2023-03-03 | ライフエディット セラピューティクス,インコーポレイティド | Rna誘導ヌクレアーゼ、その活性断片および多様体、ならびに使用方法 |
TW202208626A (zh) | 2020-04-24 | 2022-03-01 | 美商生命編輯公司 | Rna引導核酸酶及其活性片段與變體,以及使用方法 |
WO2021231437A1 (en) | 2020-05-11 | 2021-11-18 | LifeEDIT Therapeutics, Inc. | Rna-guided nucleic acid binding proteins and active fragments and variants thereof and methods of use |
AU2021310363A1 (en) | 2020-07-15 | 2023-03-16 | LifeEDIT Therapeutics, Inc. | Uracil stabilizing proteins and active fragments and variants thereof and methods of use |
AU2021339805A1 (en) | 2020-09-11 | 2023-05-25 | LifeEDIT Therapeutics, Inc. | Dna modifying enzymes and active fragments and variants thereof and methods of use |
JP2023544016A (ja) | 2020-09-30 | 2023-10-19 | パイオニア ハイ-ブレッド インターナショナル, インコーポレイテッド | 単子葉外植片の迅速な形質転換 |
US20230407324A1 (en) | 2020-10-21 | 2023-12-21 | Pioneer Hi-Bred International, Inc. | Doubled haploid inducer |
WO2022087616A1 (en) | 2020-10-21 | 2022-04-28 | Pioneer Hi-Bred International, Inc. | Parthenogenesis factors and methods of using same |
WO2022115524A2 (en) | 2020-11-24 | 2022-06-02 | AgBiome, Inc. | Pesticidal genes and methods of use |
CA3173950A1 (en) | 2021-03-22 | 2022-09-22 | Tyson D. BOWEN | Dna modifyng enzymes and active fragments and variants thereof and methods of use |
BR112023023044A2 (pt) | 2021-05-06 | 2024-01-23 | Agbiome Inc | Genes pesticidas e métodos de uso |
WO2022240931A1 (en) | 2021-05-11 | 2022-11-17 | Two Blades Foundation | Methods for preparing a library of plant disease resistance genes for functional testing for disease resistance |
EP4352215A1 (en) | 2021-06-11 | 2024-04-17 | LifeEDIT Therapeutics, Inc. | Rna polymerase iii promoters and methods of use |
CA3230261A1 (en) | 2021-09-03 | 2023-03-09 | Manuel Dubald | Plants having increased tolerance to herbicides |
AU2022356378A1 (en) | 2021-09-30 | 2024-05-02 | Two Blades Foundation | Plant disease resistance genes against stem rust and methods of use |
CA3236641A1 (en) | 2021-11-01 | 2023-05-04 | The University Of Manchester | Error prone dna polymerase for organelle mutation |
GB202116307D0 (en) | 2021-11-12 | 2021-12-29 | Syngenta Crop Protection Ag | Herbicide resistance |
CA3239251A1 (en) | 2021-12-07 | 2023-06-15 | Rebekah Deter Kelly | Pesticidal genes and methods of use |
WO2023141464A1 (en) | 2022-01-18 | 2023-07-27 | AgBiome, Inc. | Method for designing synthetic nucleotide sequences |
WO2023139557A1 (en) | 2022-01-24 | 2023-07-27 | LifeEDIT Therapeutics, Inc. | Rna-guided nucleases and active fragments and variants thereof and methods of use |
WO2023183918A1 (en) | 2022-03-25 | 2023-09-28 | Pioneer Hi-Bred International, Inc. | Methods of parthenogenic haploid induction and haploid chromosome doubling |
WO2024033901A1 (en) | 2022-08-12 | 2024-02-15 | LifeEDIT Therapeutics, Inc. | Rna-guided nucleases and active fragments and variants thereof and methods of use |
WO2024044596A1 (en) | 2022-08-23 | 2024-02-29 | AgBiome, Inc. | Pesticidal genes and methods of use |
WO2024042489A1 (en) | 2022-08-25 | 2024-02-29 | LifeEDIT Therapeutics, Inc. | Chemical modification of guide rnas with locked nucleic acid for rna guided nuclease-mediated gene editing |
WO2024052856A1 (en) | 2022-09-09 | 2024-03-14 | Friedrich Alexander Universität Erlangen-Nürnberg | Plant regulatory elements and uses thereof |
WO2024095245A2 (en) | 2022-11-04 | 2024-05-10 | LifeEDIT Therapeutics, Inc. | Evolved adenine deaminases and rna-guided nuclease fusion proteins with internal insertion sites and methods of use |
WO2024118876A1 (en) | 2022-12-01 | 2024-06-06 | Genencor International Bv | Iterative multiplex genome engineering in microbial cells using a recombinant self-excisable selection marker system |
WO2024118881A1 (en) | 2022-12-01 | 2024-06-06 | Genencor International Bv | Iterative muliplex genome engineering in microbial cells using a bidirectional selection marker system |
WO2024118882A1 (en) | 2022-12-01 | 2024-06-06 | Genencor International Bv | Iterative multiplex genome engineering in microbial cells using a selection marker swapping system |
WO2024126113A1 (en) | 2022-12-12 | 2024-06-20 | BASF Agricultural Solutions Seed US LLC | Plants having increased tolerance to herbicides |
WO2024129674A1 (en) | 2022-12-13 | 2024-06-20 | AgBiome, Inc. | Pesticidal genes and methods of use |
WO2024137438A2 (en) | 2022-12-19 | 2024-06-27 | BASF Agricultural Solutions Seed US LLC | Insect toxin genes and methods for their use |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0359472A2 (en) * | 1988-09-09 | 1990-03-21 | Mycogen Plant Science, Inc. | Synthetic insecticidal crystal protein gene |
WO1990010076A1 (en) * | 1989-02-24 | 1990-09-07 | Monsanto Company | Synthetic plant genes and method for preparation |
Family Cites Families (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
BR8404834A (pt) * | 1983-09-26 | 1985-08-13 | Agrigenetics Res Ass | Metodo para modificar geneticamente uma celula vegetal |
CA1301094C (en) * | 1984-08-31 | 1992-05-19 | Helen Riaboff Whiteley | Bacillus thuringiensis crystal protein gene toxin segment |
BR8600161A (pt) * | 1985-01-18 | 1986-09-23 | Plant Genetic Systems Nv | Gene quimerico,vetores de plasmidio hibrido,intermediario,processo para controlar insetos em agricultura ou horticultura,composicao inseticida,processo para transformar celulas de plantas para expressar uma toxina de polipeptideo produzida por bacillus thuringiensis,planta,semente de planta,cultura de celulas e plasmidio |
US4771131A (en) * | 1985-08-16 | 1988-09-13 | Mycogen Corporation | Cloning and expression of Bacillus thuringiensis toxin gene encoding a protein toxic to beetles of the order Coleoptera |
KR870004140A (ko) * | 1985-10-28 | 1987-05-07 | 모리 히데오 | 살충성 단백질 유전자의 숙주 세포내 발현에 의한 바실러스 튜린지엔시스아이자와이 ipl 아종의 살충성 단백질 제조방법 |
CA1341092C (en) * | 1985-12-12 | 2000-09-05 | David L. Edwards | Process for altering the host range of bacillus thuringiensis toxins, and novel toxins produced thereby |
DE3787350T2 (de) * | 1986-11-20 | 1994-04-21 | Monsanto Co | Insektresistente Tomatenpflanzen. |
EP0367767B1 (en) * | 1987-04-16 | 1997-03-12 | Ecogen Inc | Bacillus thuringiensis p-2 toxin gene, protein and related insecticide compositions |
TR27832A (tr) * | 1987-04-29 | 1995-08-31 | Monsanto Co | Zararli ucucu hasarata mukavim bitkiler. |
US4948734A (en) * | 1987-08-12 | 1990-08-14 | Mycogen Corporation | Novel isolates of bacillus thuringiensis having activity against nematodes |
TR24186A (tr) * | 1988-04-11 | 1991-05-30 | Monsanto Co | Hasere zehirlerinin etkinligini arttirmak icin yoentem |
EP0400246A1 (en) * | 1989-05-31 | 1990-12-05 | Plant Genetic Systems, N.V. | Prevention of Bt resistance development |
US5187091A (en) * | 1990-03-20 | 1993-02-16 | Ecogen Inc. | Bacillus thuringiensis cryiiic gene encoding toxic to coleopteran insects |
TR26973A (tr) * | 1990-04-16 | 1994-09-12 | Ecogen Inc | Bacillus thuringiensis cryie geni ve lepidoptera takimindan böceklere karsi zehirli protein. |
WO1991016432A1 (en) * | 1990-04-18 | 1991-10-31 | Plant Genetic Systems N.V. | Modified bacillus thuringiensis insecticidal-crystal protein genes and their expression in plant cells |
CA2079877C (en) * | 1990-04-26 | 2001-07-31 | Marnix Peferoen | Bacillus thuringiensis strains and their genes encoding insecticidal toxins |
WO1991018094A1 (en) * | 1990-05-24 | 1991-11-28 | Agracetus, Inc. | Particle-mediated transformation of woody plant species |
CA2042868A1 (en) * | 1990-06-11 | 1991-12-12 | Kenneth E. Narva | Bacillus thuringiensis microbes active against nematodes, and genes encoding novel nematode-active toxins cloned from bacillus thuringiensis isolates |
-
1992
- 1992-11-24 TW TW081109429A patent/TW261517B/zh active
- 1992-11-27 KR KR1019920022591A patent/KR100217517B1/ko not_active IP Right Cessation
- 1992-11-27 US US07/982,712 patent/US5436391A/en not_active Expired - Fee Related
- 1992-11-28 CN CN92115178A patent/CN1056881C/zh not_active Expired - Fee Related
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0359472A2 (en) * | 1988-09-09 | 1990-03-21 | Mycogen Plant Science, Inc. | Synthetic insecticidal crystal protein gene |
WO1990010076A1 (en) * | 1989-02-24 | 1990-09-07 | Monsanto Company | Synthetic plant genes and method for preparation |
Non-Patent Citations (3)
Title |
---|
BIO TECHNOLOGY 8 1990.1.1;BIO TECHNOLOGY 9 1991.10.1 CHRESTOU ET AL * |
BIO TECHNOLOGY 8 1990.1.1;BIO TECHNOLOGY 9 1991.10.1 CHRESTOU ET AL;SCIENCL 48 1986.1.1 * |
SCIENCL 48 1986.1.1 * |
Also Published As
Publication number | Publication date |
---|---|
US5436391A (en) | 1995-07-25 |
KR100217517B1 (ko) | 1999-10-01 |
CN1073717A (zh) | 1993-06-30 |
TW261517B (zh) | 1995-11-01 |
KR930010180A (ko) | 1993-06-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1056881C (zh) | 编码杀虫剂蛋白的基因、用该基因转化的禾本科植物及其制备方 | |
CN1250723C (zh) | 植物生长和发育的遗传调控 | |
CN1199321A (zh) | 用于在植物中防治鳞翅目昆虫的、修饰的苏云金芽胞杆菌基因 | |
CN1228784A (zh) | 编码拟南芥的gai基因的核酸 | |
CN1152961C (zh) | 控制苯丙醇生物合成通路的转录因子 | |
US20230183300A1 (en) | MODIFIED Cry1Ca TOXINS USEFUL FOR CONTROL OF INSECT PESTS | |
CN104046636B (zh) | 一种密码子植物化改造的pmi基因及其应用 | |
CN1037913C (zh) | 编码杀虫蛋白质的融合基因和表达载体及其应用 | |
CN1117867C (zh) | Dna序列及其用途 | |
CN1079779A (zh) | 新的植物启动子 | |
CN1291021C (zh) | 厚叶旋蒴苣苔BcBCP1基因在培育耐旱耐盐植物中的应用 | |
CN1824779A (zh) | 一种大豆热激转录因子及其编码基因与应用 | |
CN1226925A (zh) | 磷酸盐饥饿的诱导蛋白质 | |
CN108841833A (zh) | 一种dpbf1重组片段及其应用 | |
CN1831008A (zh) | 水稻稻瘟病菌无毒基因Avr-Pii及其应用 | |
CN100529080C (zh) | 细菌杀昆虫蛋白 | |
CN1379783A (zh) | 通过功能性地抑制植物细胞周期蛋白抑制剂基因增加植物细胞增殖的方法 | |
CN1285872A (zh) | 与植物疾病抗性相关的基因 | |
CN1184231C (zh) | 小麦TaDREB,其编码基因及培育耐逆植物的方法 | |
CN1218959C (zh) | 一种水稻乙烯受体类蛋白及其编码基因与应用 | |
CN1900280A (zh) | 水稻分蘖调控基因OsTIL1及其应用 | |
CN1793371A (zh) | 罗布麻dna解旋酶基因apdh1序列及其克隆与应用 | |
CN100351269C (zh) | 类产碱假单胞菌杀虫蛋白基因 | |
CN1860233A (zh) | 用于增加植株大小及增加叶片数目和大小的核苷酸序列及其编码的多肽 | |
CN1239705C (zh) | 一种植物耐低钾基因及其编码蛋白与应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C53 | Correction of patent for invention or patent application | ||
CB02 | Change of applicant information |
Co-applicant after: Mitsubishi Kasei Corporation Co-applicant before: Mitsubishi Kasei Corp. |
|
COR | Change of bibliographic data |
Free format text: CORRECT: CO-APPLICANT; FROM: MITSUBISHI KASEI CORP. TO: MITSUBISHI CHEMICAL CO., LTD. |
|
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
C15 | Extension of patent right duration from 15 to 20 years for appl. with date before 31.12.1992 and still valid on 11.12.2001 (patent law change 1993) | ||
OR01 | Other related matters | ||
C19 | Lapse of patent right due to non-payment of the annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |