DK2520653T3 - Integration af gener i kromosomet af saccharopolyspora spinosa - Google Patents
Integration af gener i kromosomet af saccharopolyspora spinosa Download PDFInfo
- Publication number
- DK2520653T3 DK2520653T3 DK12166602.8T DK12166602T DK2520653T3 DK 2520653 T3 DK2520653 T3 DK 2520653T3 DK 12166602 T DK12166602 T DK 12166602T DK 2520653 T3 DK2520653 T3 DK 2520653T3
- Authority
- DK
- Denmark
- Prior art keywords
- gene
- spinosyn
- spinosa
- polynucleotide
- host cell
- Prior art date
Links
- 108090000623 proteins and genes Proteins 0.000 title claims description 278
- 230000010354 integration Effects 0.000 title claims description 50
- 210000000349 chromosome Anatomy 0.000 title claims description 18
- 241000868102 Saccharopolyspora spinosa Species 0.000 title description 10
- 229930185156 spinosyn Natural products 0.000 claims description 176
- 238000000034 method Methods 0.000 claims description 136
- 230000014509 gene expression Effects 0.000 claims description 125
- 210000004027 cell Anatomy 0.000 claims description 117
- 102000040430 polynucleotide Human genes 0.000 claims description 113
- 108091033319 polynucleotide Proteins 0.000 claims description 113
- 239000002157 polynucleotide Substances 0.000 claims description 113
- 244000300264 Spinacia oleracea Species 0.000 claims description 111
- 108020004414 DNA Proteins 0.000 claims description 94
- 239000012634 fragment Substances 0.000 claims description 68
- 238000004519 manufacturing process Methods 0.000 claims description 68
- 239000013612 plasmid Substances 0.000 claims description 52
- 230000007935 neutral effect Effects 0.000 claims description 49
- 102000004190 Enzymes Human genes 0.000 claims description 48
- 108090000790 Enzymes Proteins 0.000 claims description 48
- 101710194880 Obscurin Proteins 0.000 claims description 48
- 108010030975 Polyketide Synthases Proteins 0.000 claims description 44
- 230000001851 biosynthetic effect Effects 0.000 claims description 40
- 102100030127 Obscurin Human genes 0.000 claims description 35
- 102000004169 proteins and genes Human genes 0.000 claims description 35
- 108091008053 gene clusters Proteins 0.000 claims description 28
- 108091009355 oxygen binding proteins Proteins 0.000 claims description 22
- 102000028703 oxygen binding proteins Human genes 0.000 claims description 22
- 108091026890 Coding region Proteins 0.000 claims description 18
- 239000013611 chromosomal DNA Substances 0.000 claims description 18
- 239000003550 marker Substances 0.000 claims description 17
- 230000012010 growth Effects 0.000 claims description 16
- 238000002744 homologous recombination Methods 0.000 claims description 15
- 230000006801 homologous recombination Effects 0.000 claims description 15
- 238000010367 cloning Methods 0.000 claims description 11
- 230000002068 genetic effect Effects 0.000 claims description 10
- 230000002503 metabolic effect Effects 0.000 claims description 10
- 238000011144 upstream manufacturing Methods 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 6
- 230000009466 transformation Effects 0.000 claims description 5
- 239000002609 medium Substances 0.000 claims description 4
- 238000012216 screening Methods 0.000 claims description 4
- 230000010473 stable expression Effects 0.000 claims description 4
- 230000002411 adverse Effects 0.000 claims 2
- 101150104994 PSK gene Proteins 0.000 claims 1
- 239000001963 growth medium Substances 0.000 claims 1
- 108010054147 Hemoglobins Proteins 0.000 description 58
- 150000007523 nucleic acids Chemical class 0.000 description 57
- 238000009396 hybridization Methods 0.000 description 50
- 102000039446 nucleic acids Human genes 0.000 description 50
- 108020004707 nucleic acids Proteins 0.000 description 50
- 239000000523 sample Substances 0.000 description 49
- 230000000694 effects Effects 0.000 description 39
- 238000003752 polymerase chain reaction Methods 0.000 description 36
- 108700025158 Vitreoscilla hemoglobin Proteins 0.000 description 34
- 238000002703 mutagenesis Methods 0.000 description 33
- 231100000350 mutagenesis Toxicity 0.000 description 33
- 238000000855 fermentation Methods 0.000 description 32
- 230000004151 fermentation Effects 0.000 description 32
- 102000001554 Hemoglobins Human genes 0.000 description 31
- 108090000765 processed proteins & peptides Proteins 0.000 description 31
- 239000002253 acid Substances 0.000 description 28
- 150000007513 acids Chemical class 0.000 description 27
- 230000001965 increasing effect Effects 0.000 description 26
- 239000013615 primer Substances 0.000 description 26
- 239000013598 vector Substances 0.000 description 26
- 230000003321 amplification Effects 0.000 description 23
- 238000003199 nucleic acid amplification method Methods 0.000 description 23
- 230000035772 mutation Effects 0.000 description 21
- 102000004196 processed proteins & peptides Human genes 0.000 description 21
- 230000002708 enhancing effect Effects 0.000 description 20
- 239000000047 product Substances 0.000 description 20
- 238000002741 site-directed mutagenesis Methods 0.000 description 20
- 238000004458 analytical method Methods 0.000 description 19
- 108091028043 Nucleic acid sequence Proteins 0.000 description 18
- JFLRKDZMHNBDQS-UCQUSYKYSA-N CC[C@H]1CCC[C@@H]([C@H](C(=O)C2=C[C@H]3[C@@H]4C[C@@H](C[C@H]4C(=C[C@H]3[C@@H]2CC(=O)O1)C)O[C@H]5[C@@H]([C@@H]([C@H]([C@@H](O5)C)OC)OC)OC)C)O[C@H]6CC[C@@H]([C@H](O6)C)N(C)C.CC[C@H]1CCC[C@@H]([C@H](C(=O)C2=C[C@H]3[C@@H]4C[C@@H](C[C@H]4C=C[C@H]3C2CC(=O)O1)O[C@H]5[C@@H]([C@@H]([C@H]([C@@H](O5)C)OC)OC)OC)C)O[C@H]6CC[C@@H]([C@H](O6)C)N(C)C Chemical compound CC[C@H]1CCC[C@@H]([C@H](C(=O)C2=C[C@H]3[C@@H]4C[C@@H](C[C@H]4C(=C[C@H]3[C@@H]2CC(=O)O1)C)O[C@H]5[C@@H]([C@@H]([C@H]([C@@H](O5)C)OC)OC)OC)C)O[C@H]6CC[C@@H]([C@H](O6)C)N(C)C.CC[C@H]1CCC[C@@H]([C@H](C(=O)C2=C[C@H]3[C@@H]4C[C@@H](C[C@H]4C=C[C@H]3C2CC(=O)O1)O[C@H]5[C@@H]([C@@H]([C@H]([C@@H](O5)C)OC)OC)OC)C)O[C@H]6CC[C@@H]([C@H](O6)C)N(C)C JFLRKDZMHNBDQS-UCQUSYKYSA-N 0.000 description 17
- 239000005930 Spinosad Substances 0.000 description 17
- 229920001184 polypeptide Polymers 0.000 description 17
- 229940014213 spinosad Drugs 0.000 description 17
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 16
- 238000001514 detection method Methods 0.000 description 16
- 108091034117 Oligonucleotide Proteins 0.000 description 15
- 239000000203 mixture Substances 0.000 description 15
- XZNUGFQTQHRASN-XQENGBIVSA-N apramycin Chemical compound O([C@H]1O[C@@H]2[C@H](O)[C@@H]([C@H](O[C@H]2C[C@H]1N)O[C@@H]1[C@@H]([C@@H](O)[C@H](N)[C@@H](CO)O1)O)NC)[C@@H]1[C@@H](N)C[C@@H](N)[C@H](O)[C@H]1O XZNUGFQTQHRASN-XQENGBIVSA-N 0.000 description 14
- 229950006334 apramycin Drugs 0.000 description 14
- 239000002773 nucleotide Substances 0.000 description 14
- SRJQTHAZUNRMPR-UHFFFAOYSA-N spinosyn A Natural products CC1C(=O)C2=CC3C4CC(OC5C(C(OC)C(OC)C(C)O5)OC)CC4C=CC3C2CC(=O)OC(CC)CCCC1OC1CCC(N(C)C)C(C)O1 SRJQTHAZUNRMPR-UHFFFAOYSA-N 0.000 description 14
- SRJQTHAZUNRMPR-UYQKXTDMSA-N spinosyn A Chemical compound O([C@H]1CCC[C@@H](OC(=O)C[C@H]2[C@@H]3C=C[C@@H]4C[C@H](C[C@H]4[C@@H]3C=C2C(=O)[C@@H]1C)O[C@H]1[C@@H]([C@H](OC)[C@@H](OC)[C@H](C)O1)OC)CC)[C@H]1CC[C@H](N(C)C)[C@@H](C)O1 SRJQTHAZUNRMPR-UYQKXTDMSA-N 0.000 description 14
- 238000006243 chemical reaction Methods 0.000 description 13
- 125000003729 nucleotide group Chemical group 0.000 description 13
- 229910052760 oxygen Inorganic materials 0.000 description 13
- 238000011160 research Methods 0.000 description 13
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 12
- 210000004940 nucleus Anatomy 0.000 description 12
- 239000001301 oxygen Substances 0.000 description 12
- 230000002829 reductive effect Effects 0.000 description 12
- 238000013518 transcription Methods 0.000 description 12
- 230000035897 transcription Effects 0.000 description 12
- 241000588724 Escherichia coli Species 0.000 description 11
- 150000001875 compounds Chemical class 0.000 description 11
- 230000004048 modification Effects 0.000 description 11
- 238000012986 modification Methods 0.000 description 11
- 102000053602 DNA Human genes 0.000 description 10
- 238000013459 approach Methods 0.000 description 10
- 230000015572 biosynthetic process Effects 0.000 description 10
- 238000010276 construction Methods 0.000 description 10
- 230000006798 recombination Effects 0.000 description 10
- 238000005215 recombination Methods 0.000 description 10
- 108091008146 restriction endonucleases Proteins 0.000 description 10
- 230000002441 reversible effect Effects 0.000 description 10
- JTPVHPYPNZIZKG-OWOJBTEDSA-N 5-[[4-[bis(2-hydroxyethyl)amino]-6-(4-sulfoanilino)-1,3,5-triazin-2-yl]amino]-2-[(e)-2-[4-[[4-[bis(2-hydroxyethyl)amino]-6-(4-sulfoanilino)-1,3,5-triazin-2-yl]amino]-2-sulfophenyl]ethenyl]benzenesulfonic acid Chemical compound N=1C(NC=2C=C(C(\C=C\C=3C(=CC(NC=4N=C(N=C(NC=5C=CC(=CC=5)S(O)(=O)=O)N=4)N(CCO)CCO)=CC=3)S(O)(=O)=O)=CC=2)S(O)(=O)=O)=NC(N(CCO)CCO)=NC=1NC1=CC=C(S(O)(=O)=O)C=C1 JTPVHPYPNZIZKG-OWOJBTEDSA-N 0.000 description 9
- 108020004705 Codon Proteins 0.000 description 9
- 241000196324 Embryophyta Species 0.000 description 9
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 9
- 150000001413 amino acids Chemical class 0.000 description 9
- 230000006870 function Effects 0.000 description 9
- 150000002596 lactones Chemical class 0.000 description 9
- RDECBWLKMPEKPM-UHFFFAOYSA-N spinosyn D Natural products CC1C(=O)C2=CC3C4CC(OC5C(C(OC)C(OC)C(C)O5)OC)CC4C(C)=CC3C2CC(=O)OC(CC)CCCC1OC1CCC(N(C)C)C(C)O1 RDECBWLKMPEKPM-UHFFFAOYSA-N 0.000 description 9
- RDECBWLKMPEKPM-PSCJHHPTSA-N spinosyn D Chemical compound O([C@H]1CCC[C@@H](OC(=O)C[C@H]2[C@@H]3C=C(C)[C@@H]4C[C@H](C[C@H]4[C@@H]3C=C2C(=O)[C@@H]1C)O[C@H]1[C@@H]([C@H](OC)[C@@H](OC)[C@H](C)O1)OC)CC)[C@H]1CC[C@H](N(C)C)[C@@H](C)O1 RDECBWLKMPEKPM-PSCJHHPTSA-N 0.000 description 9
- ULGZDMOVFRHVEP-RWJQBGPGSA-N Erythromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ULGZDMOVFRHVEP-RWJQBGPGSA-N 0.000 description 8
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 8
- IHGXGNJRCONUAC-WNTSOVQUSA-N Spinosyn J Chemical compound O([C@H]1CCC[C@@H](OC(=O)C[C@H]2[C@@H]3C=C[C@@H]4C[C@H](C[C@H]4[C@@H]3C=C2C(=O)[C@@H]1C)O[C@H]1[C@@H]([C@H](O)[C@@H](OC)[C@H](C)O1)OC)CC)[C@H]1CC[C@H](N(C)C)[C@@H](C)O1 IHGXGNJRCONUAC-WNTSOVQUSA-N 0.000 description 8
- 238000012217 deletion Methods 0.000 description 8
- 230000037430 deletion Effects 0.000 description 8
- 108020004999 messenger RNA Proteins 0.000 description 8
- 239000002751 oligonucleotide probe Substances 0.000 description 8
- 238000006467 substitution reaction Methods 0.000 description 8
- 235000000346 sugar Nutrition 0.000 description 8
- 238000001712 DNA sequencing Methods 0.000 description 7
- SHIBSTMRCDJXLN-UHFFFAOYSA-N Digoxigenin Natural products C1CC(C2C(C3(C)CCC(O)CC3CC2)CC2O)(O)C2(C)C1C1=CC(=O)OC1 SHIBSTMRCDJXLN-UHFFFAOYSA-N 0.000 description 7
- 102100030856 Myoglobin Human genes 0.000 description 7
- 108010062374 Myoglobin Proteins 0.000 description 7
- 241000187747 Streptomyces Species 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
- 238000007792 addition Methods 0.000 description 7
- 125000003275 alpha amino acid group Chemical group 0.000 description 7
- 238000003556 assay Methods 0.000 description 7
- 230000027455 binding Effects 0.000 description 7
- QONQRTHLHBTMGP-UHFFFAOYSA-N digitoxigenin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C11OC1CC2C1=CC(=O)OC1 QONQRTHLHBTMGP-UHFFFAOYSA-N 0.000 description 7
- 230000000670 limiting effect Effects 0.000 description 7
- -1 spnP Proteins 0.000 description 7
- 238000003786 synthesis reaction Methods 0.000 description 7
- 230000000295 complement effect Effects 0.000 description 6
- 230000021615 conjugation Effects 0.000 description 6
- 230000003247 decreasing effect Effects 0.000 description 6
- 238000000338 in vitro Methods 0.000 description 6
- 238000010369 molecular cloning Methods 0.000 description 6
- 230000010076 replication Effects 0.000 description 6
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 108700028369 Alleles Proteins 0.000 description 5
- 239000003298 DNA probe Substances 0.000 description 5
- 108700019146 Transgenes Proteins 0.000 description 5
- 238000004422 calculation algorithm Methods 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 230000002759 chromosomal effect Effects 0.000 description 5
- 238000012790 confirmation Methods 0.000 description 5
- 230000029087 digestion Effects 0.000 description 5
- SHIBSTMRCDJXLN-KCZCNTNESA-N digoxigenin Chemical compound C1([C@@H]2[C@@]3([C@@](CC2)(O)[C@H]2[C@@H]([C@@]4(C)CC[C@H](O)C[C@H]4CC2)C[C@H]3O)C)=CC(=O)OC1 SHIBSTMRCDJXLN-KCZCNTNESA-N 0.000 description 5
- 102000018146 globin Human genes 0.000 description 5
- 108060003196 globin Proteins 0.000 description 5
- 230000003116 impacting effect Effects 0.000 description 5
- 230000002779 inactivation Effects 0.000 description 5
- 230000037353 metabolic pathway Effects 0.000 description 5
- 238000007899 nucleic acid hybridization Methods 0.000 description 5
- 230000008685 targeting Effects 0.000 description 5
- 101150020820 vhb gene Proteins 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 108020003215 DNA Probes Proteins 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 101710187052 Flavohemoprotein Proteins 0.000 description 4
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- 241000238631 Hexapoda Species 0.000 description 4
- 108010063653 Leghemoglobin Proteins 0.000 description 4
- 108020004711 Nucleic Acid Probes Proteins 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 4
- 238000003776 cleavage reaction Methods 0.000 description 4
- 238000006911 enzymatic reaction Methods 0.000 description 4
- 229960003276 erythromycin Drugs 0.000 description 4
- 239000013604 expression vector Substances 0.000 description 4
- 238000010353 genetic engineering Methods 0.000 description 4
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 4
- 238000003780 insertion Methods 0.000 description 4
- 230000037431 insertion Effects 0.000 description 4
- 239000000543 intermediate Substances 0.000 description 4
- 238000002372 labelling Methods 0.000 description 4
- 238000007834 ligase chain reaction Methods 0.000 description 4
- 238000011068 loading method Methods 0.000 description 4
- 239000011159 matrix material Substances 0.000 description 4
- 230000001404 mediated effect Effects 0.000 description 4
- 239000002853 nucleic acid probe Substances 0.000 description 4
- 230000002018 overexpression Effects 0.000 description 4
- 239000002243 precursor Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 230000003362 replicative effect Effects 0.000 description 4
- 230000007017 scission Effects 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 241001515965 unidentified phage Species 0.000 description 4
- INGWEZCOABYORO-UHFFFAOYSA-N 2-(furan-2-yl)-7-methyl-1h-1,8-naphthyridin-4-one Chemical compound N=1C2=NC(C)=CC=C2C(O)=CC=1C1=CC=CO1 INGWEZCOABYORO-UHFFFAOYSA-N 0.000 description 3
- 241000186046 Actinomyces Species 0.000 description 3
- 241000203809 Actinomycetales Species 0.000 description 3
- VTIKDEXOEJDMJP-UHFFFAOYSA-N Actinorhodine Natural products CC1OC(CC(=O)O)CC2=C1C(=O)c3c(O)c(cc(O)c3C2=O)c4cc(O)c5C(=O)C6=C(C(C)OC(CC(=O)O)C6)C(=O)c5c4O VTIKDEXOEJDMJP-UHFFFAOYSA-N 0.000 description 3
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 3
- 230000004544 DNA amplification Effects 0.000 description 3
- 238000007399 DNA isolation Methods 0.000 description 3
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 3
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 3
- 108091005461 Nucleic proteins Chemical group 0.000 description 3
- 108700026244 Open Reading Frames Proteins 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- VTIKDEXOEJDMJP-WYUUTHIRSA-N actinorhodin Chemical compound C([C@@H](CC(O)=O)O[C@@H]1C)C(C(C2=C(O)C=3)=O)=C1C(=O)C2=C(O)C=3C(C(=C1C2=O)O)=CC(O)=C1C(=O)C1=C2[C@@H](C)O[C@H](CC(O)=O)C1 VTIKDEXOEJDMJP-WYUUTHIRSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 230000003115 biocidal effect Effects 0.000 description 3
- 239000007795 chemical reaction product Substances 0.000 description 3
- 150000008266 deoxy sugars Chemical class 0.000 description 3
- 230000012361 double-strand break repair Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- SZGAAHDUAFVZSS-SFYZADRCSA-N forosamine Chemical compound C[C@@H](O)[C@@H](N(C)C)CCC=O SZGAAHDUAFVZSS-SFYZADRCSA-N 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 239000002917 insecticide Substances 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000003757 reverse transcription PCR Methods 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- SRJQTHAZUNRMPR-SFGMUSMWSA-N spinosyn-a Chemical compound C([C@@H](OC(=O)C[C@H]1[C@@H]2C=C[C@@H]3C[C@H](C[C@H]3[C@@H]2C=C1C(=O)[C@@H]1C)O[C@H]2[C@@H]([C@H](OC)[C@@H](OC)[C@H](C)O2)OC)CC)CCC1O[C@H]1CC[C@H](N(C)C)[C@@H](C)O1 SRJQTHAZUNRMPR-SFGMUSMWSA-N 0.000 description 3
- 238000006257 total synthesis reaction Methods 0.000 description 3
- QRXMUCSWCMTJGU-UHFFFAOYSA-L (5-bromo-4-chloro-1h-indol-3-yl) phosphate Chemical compound C1=C(Br)C(Cl)=C2C(OP([O-])(=O)[O-])=CNC2=C1 QRXMUCSWCMTJGU-UHFFFAOYSA-L 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 2
- 241000239223 Arachnida Species 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 241000252867 Cupriavidus metallidurans Species 0.000 description 2
- 102100028717 Cytosolic 5'-nucleotidase 3A Human genes 0.000 description 2
- 238000000018 DNA microarray Methods 0.000 description 2
- 239000003155 DNA primer Substances 0.000 description 2
- 230000004568 DNA-binding Effects 0.000 description 2
- 241000255925 Diptera Species 0.000 description 2
- 108060002716 Exonuclease Proteins 0.000 description 2
- 102000034286 G proteins Human genes 0.000 description 2
- 108091006027 G proteins Proteins 0.000 description 2
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- 102000004867 Hydro-Lyases Human genes 0.000 description 2
- 108090001042 Hydro-Lyases Proteins 0.000 description 2
- 101001110310 Lentilactobacillus kefiri NADP-dependent (R)-specific alcohol dehydrogenase Proteins 0.000 description 2
- 241000255777 Lepidoptera Species 0.000 description 2
- 241000219745 Lupinus Species 0.000 description 2
- 101001014220 Monascus pilosus Dehydrogenase mokE Proteins 0.000 description 2
- 241000244206 Nematoda Species 0.000 description 2
- 238000000636 Northern blotting Methods 0.000 description 2
- 101000573542 Penicillium citrinum Compactin nonaketide synthase, enoyl reductase component Proteins 0.000 description 2
- 241000283222 Physeter catodon Species 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- 102000018120 Recombinases Human genes 0.000 description 2
- 108010091086 Recombinases Proteins 0.000 description 2
- 241000187559 Saccharopolyspora erythraea Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000005929 Spinetoram Substances 0.000 description 2
- GOENIMGKWNZVDA-OAMCMWGQSA-N Spinetoram Chemical compound CO[C@@H]1[C@H](OCC)[C@@H](OC)[C@H](C)O[C@H]1OC1C[C@H]2[C@@H]3C=C4C(=O)[C@H](C)[C@@H](O[C@@H]5O[C@H](C)[C@H](CC5)N(C)C)CCC[C@H](CC)OC(=O)CC4[C@@H]3CC[C@@H]2C1 GOENIMGKWNZVDA-OAMCMWGQSA-N 0.000 description 2
- 108091081024 Start codon Proteins 0.000 description 2
- RYYWUUFWQRZTIU-UHFFFAOYSA-N Thiophosphoric acid Chemical group OP(O)(S)=O RYYWUUFWQRZTIU-UHFFFAOYSA-N 0.000 description 2
- 102000006612 Transducin Human genes 0.000 description 2
- 108010087042 Transducin Proteins 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 230000004075 alteration Effects 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 108010065885 aminoglycoside N(3')-acetyltransferase Proteins 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 239000012472 biological sample Substances 0.000 description 2
- 230000006696 biosynthetic metabolic pathway Effects 0.000 description 2
- 238000006664 bond formation reaction Methods 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 2
- 238000012219 cassette mutagenesis Methods 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000001976 enzyme digestion Methods 0.000 description 2
- 102000013165 exonuclease Human genes 0.000 description 2
- 239000013613 expression plasmid Substances 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 230000030279 gene silencing Effects 0.000 description 2
- 238000012226 gene silencing method Methods 0.000 description 2
- 238000012252 genetic analysis Methods 0.000 description 2
- 150000002337 glycosamines Chemical class 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000000749 insecticidal effect Effects 0.000 description 2
- 125000000468 ketone group Chemical group 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000003120 macrolide antibiotic agent Substances 0.000 description 2
- 238000012269 metabolic engineering Methods 0.000 description 2
- 230000033607 mismatch repair Effects 0.000 description 2
- 231100000219 mutagenic Toxicity 0.000 description 2
- 230000003505 mutagenic effect Effects 0.000 description 2
- MHWLWQUZZRMNGJ-UHFFFAOYSA-N nalidixic acid Chemical compound C1=C(C)N=C2N(CC)C=C(C(O)=O)C(=O)C2=C1 MHWLWQUZZRMNGJ-UHFFFAOYSA-N 0.000 description 2
- 229960000210 nalidixic acid Drugs 0.000 description 2
- JPXMTWWFLBLUCD-UHFFFAOYSA-N nitro blue tetrazolium(2+) Chemical compound COC1=CC(C=2C=C(OC)C(=CC=2)[N+]=2N(N=C(N=2)C=2C=CC=CC=2)C=2C=CC(=CC=2)[N+]([O-])=O)=CC=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=C([N+]([O-])=O)C=C1 JPXMTWWFLBLUCD-UHFFFAOYSA-N 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- RGCLLPNLLBQHPF-HJWRWDBZSA-N phosphamidon Chemical compound CCN(CC)C(=O)C(\Cl)=C(/C)OP(=O)(OC)OC RGCLLPNLLBQHPF-HJWRWDBZSA-N 0.000 description 2
- 229930001119 polyketide Natural products 0.000 description 2
- 230000032361 posttranscriptional gene silencing Effects 0.000 description 2
- 230000037452 priming Effects 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 235000008001 rakum palm Nutrition 0.000 description 2
- 238000002708 random mutagenesis Methods 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 108010066533 ribonuclease S Proteins 0.000 description 2
- 210000004358 rod cell outer segment Anatomy 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- 230000000087 stabilizing effect Effects 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 230000007704 transition Effects 0.000 description 2
- ZUGIGWIPEYNYJV-FZWPAWTOSA-N (3r,4s,5r,6s,7s,9r,11r,12r,13s,14r)-6-[(2s,3r,4s,6r)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy-14-ethyl-12,13-dihydroxy-4-[(2r,4r,5s,6s)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy-3,5,7,9,11,13-hexamethyl-oxacyclotetradecane-2,10-dione Chemical class O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@H](C)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ZUGIGWIPEYNYJV-FZWPAWTOSA-N 0.000 description 1
- YKAVHPRGGAUFDN-JTQLBUQXSA-N 24464-30-0 Chemical class O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]2(C)O[C@]3([C@@H]([C@H]2O)C)[C@H](C)C[C@](O3)(C)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 YKAVHPRGGAUFDN-JTQLBUQXSA-N 0.000 description 1
- 101000715489 Acinetobacter lwoffii Probable cat1 operon transcriptional activator Proteins 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 244000105975 Antidesma platyphyllum Species 0.000 description 1
- 108020005544 Antisense RNA Proteins 0.000 description 1
- 101000886924 Aplysia limacina Globin Proteins 0.000 description 1
- 108010085614 Ascaris hemoglobin Proteins 0.000 description 1
- 241000972773 Aulopiformes Species 0.000 description 1
- 101100311670 Bacillus subtilis (strain 168) cysS gene Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108010077805 Bacterial Proteins Proteins 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 239000004099 Chlortetracycline Substances 0.000 description 1
- 108091033380 Coding strand Proteins 0.000 description 1
- 108700010070 Codon Usage Proteins 0.000 description 1
- 108020004635 Complementary DNA Proteins 0.000 description 1
- 102100031673 Corneodesmosin Human genes 0.000 description 1
- 241001464430 Cyanobacterium Species 0.000 description 1
- 102000006465 DNA Restriction-Modification Enzymes Human genes 0.000 description 1
- 108010044289 DNA Restriction-Modification Enzymes Proteins 0.000 description 1
- 230000008836 DNA modification Effects 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 101100310822 Dictyostelium discoideum spnA gene Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 241000078280 Escherichia coli S17 Species 0.000 description 1
- QTANTQQOYSUMLC-UHFFFAOYSA-O Ethidium cation Chemical compound C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 QTANTQQOYSUMLC-UHFFFAOYSA-O 0.000 description 1
- CEAGUSGLAUVBEQ-UHFFFAOYSA-N Forosamine Natural products CC1CC(N(C)C)CC(O)O1 CEAGUSGLAUVBEQ-UHFFFAOYSA-N 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 102000012330 Integrases Human genes 0.000 description 1
- 108010061833 Integrases Proteins 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- 150000008575 L-amino acids Chemical class 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- 102000016397 Methyltransferase Human genes 0.000 description 1
- 108091092878 Microsatellite Proteins 0.000 description 1
- 229930191564 Monensin Natural products 0.000 description 1
- GAOZTHIDHYLHMS-UHFFFAOYSA-N Monensin A Natural products O1C(CC)(C2C(CC(O2)C2C(CC(C)C(O)(CO)O2)C)C)CCC1C(O1)(C)CCC21CC(O)C(C)C(C(C)C(OC)C(C)C(O)=O)O2 GAOZTHIDHYLHMS-UHFFFAOYSA-N 0.000 description 1
- BACYUWVYYTXETD-UHFFFAOYSA-N N-Lauroylsarcosine Chemical compound CCCCCCCCCCCC(=O)N(C)CC(O)=O BACYUWVYYTXETD-UHFFFAOYSA-N 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 101000649053 Paramecium caudatum Group 1 truncated hemoglobin trHbN Proteins 0.000 description 1
- 241000364057 Peoria Species 0.000 description 1
- 241000244041 Pseudoterranova decipiens Species 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 101100422141 Saccharopolyspora spinosa spnN gene Proteins 0.000 description 1
- 101100422142 Saccharopolyspora spinosa spnO gene Proteins 0.000 description 1
- 101100422143 Saccharopolyspora spinosa spnQ gene Proteins 0.000 description 1
- 101100422144 Saccharopolyspora spinosa spnR gene Proteins 0.000 description 1
- 101100366459 Saccharopolyspora spinosa spnS gene Proteins 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241000237506 Scapharca Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- 241000589196 Sinorhizobium meliloti Species 0.000 description 1
- 238000002105 Southern blotting Methods 0.000 description 1
- JVXKZYLRDBNKCL-KXRJSVEISA-N Spinosyn K Chemical compound O([C@H]1CCC[C@@H](OC(=O)C[C@H]2[C@@H]3C=C[C@@H]4C[C@H](C[C@H]4[C@@H]3C=C2C(=O)[C@@H]1C)O[C@H]1[C@@H]([C@H](OC)[C@@H](O)[C@H](C)O1)OC)CC)[C@H]1CC[C@H](N(C)C)[C@@H](C)O1 JVXKZYLRDBNKCL-KXRJSVEISA-N 0.000 description 1
- 241000187432 Streptomyces coelicolor Species 0.000 description 1
- 241000187438 Streptomyces fradiae Species 0.000 description 1
- 108090000787 Subtilisin Proteins 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- 241001414989 Thysanoptera Species 0.000 description 1
- 229930194936 Tylosin Natural products 0.000 description 1
- 239000004182 Tylosin Substances 0.000 description 1
- CSCPPACGZOOCGX-WFGJKAKNSA-N acetone d6 Chemical compound [2H]C([2H])([2H])C(=O)C([2H])([2H])[2H] CSCPPACGZOOCGX-WFGJKAKNSA-N 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 238000002820 assay format Methods 0.000 description 1
- 238000000376 autoradiography Methods 0.000 description 1
- QVZZPLDJERFENQ-NKTUOASPSA-N bassianolide Chemical compound CC(C)C[C@@H]1N(C)C(=O)[C@@H](C(C)C)OC(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C(C)C)OC(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C(C)C)OC(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C(C)C)OC1=O QVZZPLDJERFENQ-NKTUOASPSA-N 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- CYDMQBQPVICBEU-UHFFFAOYSA-N chlorotetracycline Natural products C1=CC(Cl)=C2C(O)(C)C3CC4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-UHFFFAOYSA-N 0.000 description 1
- 229960004475 chlortetracycline Drugs 0.000 description 1
- 235000019365 chlortetracycline Nutrition 0.000 description 1
- CYDMQBQPVICBEU-XRNKAMNCSA-N chlortetracycline Chemical compound C1=CC(Cl)=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-XRNKAMNCSA-N 0.000 description 1
- 230000004186 co-expression Effects 0.000 description 1
- 239000003184 complementary RNA Substances 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013601 cosmid vector Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000000368 destabilizing effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000009088 enzymatic function Effects 0.000 description 1
- 238000009585 enzyme analysis Methods 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 229960005542 ethidium bromide Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 238000003208 gene overexpression Methods 0.000 description 1
- 235000003869 genetically modified organism Nutrition 0.000 description 1
- 230000008826 genomic mutation Effects 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 239000011544 gradient gel Substances 0.000 description 1
- 235000009424 haa Nutrition 0.000 description 1
- 238000003505 heat denaturation Methods 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 238000005984 hydrogenation reaction Methods 0.000 description 1
- 230000001146 hypoxic effect Effects 0.000 description 1
- 230000014726 immortalization of host cell Effects 0.000 description 1
- 229940127121 immunoconjugate Drugs 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011005 laboratory method Methods 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 108010026228 mRNA guanylyltransferase Proteins 0.000 description 1
- UFUYRGNJTFAODM-HQCAVAADSA-N macrocin Chemical compound O([C@@H]1[C@@H](C)O[C@H]([C@@H]([C@H]1N(C)C)O)O[C@@H]1[C@@H](C)[C@H](O)CC(=O)O[C@@H]([C@H](/C=C(\C)/C=C/C(=O)[C@H](C)C[C@@H]1CC=O)CO[C@H]1[C@@H]([C@H](O)[C@H](O)[C@@H](C)O1)OC)CC)[C@H]1C[C@@](C)(O)[C@@H](O)[C@H](C)O1 UFUYRGNJTFAODM-HQCAVAADSA-N 0.000 description 1
- 229940041033 macrolides Drugs 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 229960005358 monensin Drugs 0.000 description 1
- GAOZTHIDHYLHMS-KEOBGNEYSA-N monensin A Chemical compound C([C@@](O1)(C)[C@H]2CC[C@@](O2)(CC)[C@H]2[C@H](C[C@@H](O2)[C@@H]2[C@H](C[C@@H](C)[C@](O)(CO)O2)C)C)C[C@@]21C[C@H](O)[C@@H](C)[C@@H]([C@@H](C)[C@@H](OC)[C@H](C)C(O)=O)O2 GAOZTHIDHYLHMS-KEOBGNEYSA-N 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 230000004108 pentose phosphate pathway Effects 0.000 description 1
- 239000000816 peptidomimetic Substances 0.000 description 1
- JTJMJGYZQZDUJJ-UHFFFAOYSA-N phencyclidine Chemical compound C1CCCCN1C1(C=2C=CC=CC=2)CCCCC1 JTJMJGYZQZDUJJ-UHFFFAOYSA-N 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000019525 primary metabolic process Effects 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 239000000700 radioactive tracer Substances 0.000 description 1
- 238000009790 rate-determining step (RDS) Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 125000006853 reporter group Chemical group 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 235000019515 salmon Nutrition 0.000 description 1
- 108700004121 sarkosyl Proteins 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000010187 selection method Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 231100000583 toxicological profile Toxicity 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000017105 transposition Effects 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- 238000013024 troubleshooting Methods 0.000 description 1
- WBPYTXDJUQJLPQ-VMXQISHHSA-N tylosin Chemical compound O([C@@H]1[C@@H](C)O[C@H]([C@@H]([C@H]1N(C)C)O)O[C@@H]1[C@@H](C)[C@H](O)CC(=O)O[C@@H]([C@H](/C=C(\C)/C=C/C(=O)[C@H](C)C[C@@H]1CC=O)CO[C@H]1[C@@H]([C@H](OC)[C@H](O)[C@@H](C)O1)OC)CC)[C@H]1C[C@@](C)(O)[C@@H](O)[C@H](C)O1 WBPYTXDJUQJLPQ-VMXQISHHSA-N 0.000 description 1
- 229960004059 tylosin Drugs 0.000 description 1
- 235000019375 tylosin Nutrition 0.000 description 1
- 230000009452 underexpressoin Effects 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
- C12P19/62—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin the hetero ring having eight or more ring members and only oxygen as ring hetero atoms, e.g. erythromycin, spiramycin, nystatin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
Landscapes
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Claims (24)
1. Fremgangsmåde til integration af et gen ved homolog rekombination i kromosomalt DNA af en S. spinosa-værtscelle, hvilken fremgangsmåde omfatter trinnene: a) at klone to genomiske DNA-fragmenter fra obsA-genet af obscurin-po-lyketidsynthase-genlocuset for at frembringe et integrationsplasmid, hvilket ene genomiske fragment klones opstrøms for et polynukleotid, der skal integreres, og det andet genomiske fragment klones nedstrøms for et polynukleotid, der skal integreres, og det resulterende ligerede DNA-fragment bliver integrationsplasmidet, hvilket obsA-gen af obscurin-poly-ketidsynthase-genlocuset ved integration afbrydes af polynukleotidet; b) at indføre plasmidet fra trin (a) ind i værtscellen; c) at integrere polynukleotidet i det native obsA-gen af obscurin-polyke-tidsynthase-genlocuset i genomet af værtscellen; og d) at screene værtscellen for tilstedeværelse af polynukleotidet.
2. Fremgangsmåden ifølge krav 1, hvor integration af polynukleotidet ikke påvirker vækst af S. spinosa eller en ønsket metabolisk karakteristik negativt.
3. Fremgangsmåde til stabil ekspression af et polynukleotid i en S. spinosa-værtscelle, der omfatter en fremgangsmåde ifølge krav 1 eller 2.
4. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 3, hvor polynukleotidet omfatter en genekspressionskassette, hvilken genekspressionskassette omfatter et gen, der koder for et spinosyn-biosyntetisk enzym og/eller et gen, der koder for et oxygen-bindende protein.
5. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 4, hvor polynukleotidet omfatter en selekterbar markør.
6. Genetisk modificeret S. spinosa-værtscelle, der omfatter et tilsat gen i det kromosomale DNA ved PSK-genet obsA for obscurin-polyketidsynthase-gen-klyngen eller et substitueret gen, som ikke er identisk med et nativt S. spinosa-gen.
7. Genetisk modificeret S. spinosa-værtscelle ifølge krav 6, hvor genet, som er blevet tilføjet, er identisk med et nativt S. spinosa-gen.
8. Fremgangsmåde til fremstilling af et proteinprodukt af et target-gen, hvilken fremgangsmåde omfatter at fremstille en S. sp/nosa-værtcelle, i hvilken et bestemt antal kopier af target-genet er integreret i genomet af S. spinosa ifølge trin (a) til (d) i krav 1, og at dyrke cellen i et egnet dyrkningsmedium i tilstrækkelig tid for target-genet til at blive udtrykt og for det nævnte protein til at blive produceret.
9. Fremgangsmåde til stabil ekspression af et target-gen, hvilken fremgangsmåde omfatter: at integrere en obscurin-polyketidsyntase-genklynge i et S. spinosa-genom som et neutralt site for integration og ekspression af et target-gen af interesse; at integrere target-genet af interesse i det neutrale site, hvorved obscu-rin-polyketidsynthase-genklyngen afbrydes; hvilket target-gen af interesse koder for et oxygen-bindende protein.
10. Fremgangsmåden ifølge krav 9, hvor endvidere obscurin-PKS-genet obsA af den integrerede klynge er stedet for integration.
11. Fremgangsmåde ifølge krav 9 eller 10, hvor target-genet omfatter en kodende sekvens, der er funktionelt forbundet til en heterolog promotor, der er heterolog til stamme af S. spinosa.
12. Fremgangsmåde ifølge ethvert af kravene 9 til 11, hvor target-genet omfatter en selekterbar markør.
13. Fremgangsmåde ifølge ethvert af kravene 9 til 12, hvor integrationen omfatter homolog rekombination.
14. Fremgangsmåde ifølge et hvilket som helst af kravene 9 til 13, hvilken fremgangsmåde yderligere omfatter: at dyrke en værtscelle under betingelser, der passer til fremstilling af spi-nosyn; og at opsamle spinosyn fra en kultur af værtscellen.
15. Ekspressionskonstrukt, der omfatter obscurin-polyketidsyntase-genet obsA, hvor obsA-genet omfatter et gen, der koder for et oxygen-bindingsprotein, og som er integreret deri.
16. Ekspressionskonstrukt ifølge krav 15, hvor ekspressionskonstruktet er egnet til integration i et værtscellegenom.
17. Anvendelse af et ekspressionskonstrukt ifølge krav 15 eller 16 i en fremgangsmåde til integration af et gen ved homolog rekombination i kromosomalt DNA af en S. spinosa-værtscelle til stabil genekspression.
18. Fremgangsmåde til fremstilling af en S. spinosa stamme, i hvilken et bestemt antal kopier af et udvalgt gen eller polynukleotid er integreret i et S. spi-nosa-genom, hvilken fremgangsmåde omfatter: a) at klone et obscurin-polyketid-syntase-genlocus til et mobilt element, i hvilket der er mindst ét markørgen, som er placeret inden i det mobile element; b) at integrere det mobile element i et obscurin-polyketidsyntase-genlo-cus i kromosomet eller episomet af den nævnte S. spinosa-stamme; c) at komplementere det nævnte mobile element med et funktionelt gen; d) at udtrykke det nævnte funktionelle gen af det mobile element; e) at udvælge stammen på et medium, der er selekterbart for det nævnte markørgen på en sådan måde, at der opnås en stamme, der indeholder et bestemt antal kopier af obscurin-polyketidsynthase-genlocuset; f) at udvinde den derved udvalgte stamme; og g) at integrere et polynukleotid i det specifikke antal kopier af obscurin-polyketid-synthase-genlocuset ved obscurin-PKS-genet obsA, hvorved obscurin-polyketidsynthase-genlocuset afbrydes.
19. Fremgangsmåden ifølge krav 18, hvor kromosomet eller episomet er indeholdt på et plasmid og indføres ved transformation.
20. Fremgangsmåden ifølge krav 18 eller 19, hvor det mobile element er et genetisk konstrukt.
21. Fremgangsmåde, celle eller konstrukt ifølge ethvert af de foregående krav, hvor integration af et polynukleotid ikke negativ påvirker produktion, vækst eller andre ønskede metaboliske egenskaber af spinosyn.
22. Anvendelse af en genetisk modificeret S. sp/nosa-værtcelle, der omfatter et tilsat, substitueret eller deleteret gen i det kromosomale DNA i obscurin-polyke-tidsyntase-genklyngen, i en fremgangsmåde til stabil genekspression.
23. Anvendelse ifølge krav 22, hvor værtscellen er som defineret i ethvert af kravene 6 eller 7.
24. Anvendelse ifølge krav 22 eller 23, hvor fremgangsmåden omfatter at dyrke værtscellen under betingelser, der passer for fremstilling af spinosyn; og for opsamling af spinosyn fra en kultur af værtscellen.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US13/100,202 US8741603B2 (en) | 2011-05-03 | 2011-05-03 | Enhancing spinosyn production with oxygen binding proteins |
| US13/100,220 US9404107B2 (en) | 2011-05-03 | 2011-05-03 | Integration of genes into the chromosome of Saccharopolyspora spinosa |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2520653T3 true DK2520653T3 (da) | 2017-07-03 |
Family
ID=46001043
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK12166602.8T DK2520653T3 (da) | 2011-05-03 | 2012-05-03 | Integration af gener i kromosomet af saccharopolyspora spinosa |
Country Status (4)
| Country | Link |
|---|---|
| EP (3) | EP2998399A1 (da) |
| DK (1) | DK2520653T3 (da) |
| ES (2) | ES2625504T3 (da) |
| HU (1) | HUE034838T2 (da) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2513859B (en) * | 2013-05-07 | 2018-01-17 | Rotam Agrochem Int Co Ltd | Agrochemical composition, method for its preparation and the use thereof |
| WO2018226893A2 (en) * | 2017-06-06 | 2018-12-13 | Zymergen Inc. | A high-throughput (htp) genomic engineering platform for improving saccharopolyspora spinosa |
| CN107418925B (zh) * | 2017-06-13 | 2020-03-31 | 国家粮食和物资储备局科学研究院 | 多杀菌素高产基因工程菌及其构建方法和应用 |
| CN110511990A (zh) * | 2019-09-12 | 2019-11-29 | 广东美立康生物科技有限公司 | 一种棘胸蛙蝌蚪变态期死亡风险预测技术 |
| CN110656093A (zh) * | 2019-10-12 | 2020-01-07 | 山东省农业科学院畜牧兽医研究所 | 一种新型沙门氏菌噬菌体库及其应用 |
| CN114773454B (zh) * | 2022-04-27 | 2022-11-29 | 广州蕊特生物科技有限公司 | 从马心中提取肌红蛋白的提取与纯化方法 |
Family Cites Families (29)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4302204A (en) | 1979-07-02 | 1981-11-24 | The Board Of Trustees Of Leland Stanford Junior University | Transfer and detection of nucleic acids |
| US4358535A (en) | 1980-12-08 | 1982-11-09 | Board Of Regents Of The University Of Washington | Specific DNA probes in diagnostic microbiology |
| FI63596C (fi) | 1981-10-16 | 1983-07-11 | Orion Yhtymae Oy | Mikrobdiagnostiskt foerfarande som grundar sig pao skiktshybridisering av nukleinsyror och vid foerfarandet anvaenda kombinationer av reagenser |
| US4994373A (en) | 1983-01-27 | 1991-02-19 | Enzo Biochem, Inc. | Method and structures employing chemically-labelled polynucleotide probes |
| US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
| US4683195A (en) | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
| US4965188A (en) | 1986-08-22 | 1990-10-23 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme |
| EP0228075B1 (en) | 1986-01-03 | 1991-04-03 | Molecular Diagnostics, Inc. | Eucaryotic genomic dna dot-blot hybridization method |
| US4800159A (en) | 1986-02-07 | 1989-01-24 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences |
| US5202231A (en) | 1987-04-01 | 1993-04-13 | Drmanac Radoje T | Method of sequencing of genomes by hybridization of oligonucleotide probes |
| IL88095A0 (en) * | 1987-10-23 | 1989-06-30 | California Inst Of Techn | Dna sequences,proteins and gene preparations |
| US5049493A (en) | 1987-10-23 | 1991-09-17 | California Institute Of Technology | Enhancement of cell growth by expression of a cloned hemoglobin gene |
| US5030557A (en) | 1987-11-24 | 1991-07-09 | Ml Technology Venture | Means and method for enhancing nucleic acid hybridization |
| OA09249A (fr) | 1988-12-19 | 1992-06-30 | Lilly Co Eli | Composés de macrolides. |
| US5362634A (en) | 1989-10-30 | 1994-11-08 | Dowelanco | Process for producing A83543 compounds |
| US5231020A (en) | 1989-03-30 | 1993-07-27 | Dna Plant Technology Corporation | Genetic engineering of novel plant phenotypes |
| US5034323A (en) | 1989-03-30 | 1991-07-23 | Dna Plant Technology Corporation | Genetic engineering of novel plant phenotypes |
| US5219726A (en) | 1989-06-02 | 1993-06-15 | The Salk Institute For Biological Studies | Physical mapping of complex genomes |
| CN1073483A (zh) | 1991-11-08 | 1993-06-23 | 道伊兰科公司 | 一种发酵杀虫剂化合物及其制备方法 |
| WO1994020518A1 (en) | 1993-03-12 | 1994-09-15 | Dowelanco | New a83543 compounds and process for production thereof |
| US5770365A (en) | 1995-08-25 | 1998-06-23 | Tm Technologies, Inc. | Nucleic acid capture moieties |
| US6001981A (en) | 1996-06-13 | 1999-12-14 | Dow Agrosciences Llc | Synthetic modification of Spinosyn compounds |
| US6506559B1 (en) | 1997-12-23 | 2003-01-14 | Carnegie Institute Of Washington | Genetic inhibition by double-stranded RNA |
| US6143526A (en) | 1998-03-09 | 2000-11-07 | Baltz; Richard H. | Biosynthetic genes for spinosyn insecticide production |
| CN1202246C (zh) | 1998-04-08 | 2005-05-18 | 联邦科学和工业研究组织 | 获得修饰表型的方法和措施 |
| US7285653B1 (en) * | 1999-08-27 | 2007-10-23 | Bayer Aktiengesellschaft | Nucleic acids which code for the enzyme activities of the spinosyn biosynthesis |
| AR033022A1 (es) * | 2001-03-30 | 2003-12-03 | Dow Agrosciences Llc | Genes biosinteticos para la produccion de insecticida de butenilespinosina |
| IL163529A0 (en) | 2002-02-19 | 2005-12-18 | Dow Agrosciences Llc | Novel spinosyn-producing polyketidesynthases |
| US7996021B2 (en) | 2002-06-28 | 2011-08-09 | Hewlett-Packard Development Company, L.P. | Location determination in a wireless communication network |
-
2012
- 2012-05-03 DK DK12166602.8T patent/DK2520653T3/da active
- 2012-05-03 EP EP15182915.7A patent/EP2998399A1/en not_active Withdrawn
- 2012-05-03 EP EP12166602.8A patent/EP2520653B1/en active Active
- 2012-05-03 ES ES12166602.8T patent/ES2625504T3/es active Active
- 2012-05-03 HU HUE12166602A patent/HUE034838T2/en unknown
- 2012-05-03 EP EP12166535.0A patent/EP2520658B1/en active Active
- 2012-05-03 ES ES12166535.0T patent/ES2553079T3/es active Active
Also Published As
| Publication number | Publication date |
|---|---|
| ES2625504T3 (es) | 2017-07-19 |
| ES2553079T3 (es) | 2015-12-04 |
| EP2520653B1 (en) | 2017-03-29 |
| HUE034838T2 (en) | 2018-03-28 |
| EP2520658A2 (en) | 2012-11-07 |
| EP2520653A1 (en) | 2012-11-07 |
| EP2520658A3 (en) | 2013-02-20 |
| EP2998399A1 (en) | 2016-03-23 |
| EP2520658B1 (en) | 2015-10-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20140248667A1 (en) | Enhancing spinosyn production with oxygen binding proteins | |
| DK2520653T3 (da) | Integration af gener i kromosomet af saccharopolyspora spinosa | |
| CA2322449C (en) | Biosynthetic genes for spinosyn insecticide production | |
| CA2197160C (en) | Platenolide synthase gene | |
| EP2271666A1 (en) | Nrps-pks gene cluster and its manipulation and utility | |
| CA2332129A1 (en) | Dna encoding methymycin and pikromycin | |
| US9404107B2 (en) | Integration of genes into the chromosome of Saccharopolyspora spinosa | |
| CN103119152B (zh) | Spnk菌株 | |
| CN101363022B (zh) | 替曲卡星a的生物合成基因簇及其应用 | |
| EP2610344B1 (en) | Identification and characterization of the spinactin biosynthesis gene cluster from spinosyn-producing Saccharopolyspora spinosa | |
| CN100465277C (zh) | 氯丝菌素的生物合成基因簇及其应用 | |
| CN116515876A (zh) | 异源合成黄酮类化合物的调控方法与应用 | |
| WO2009147984A1 (ja) | ハーボキシジエンの生合成に関与するポリペプチドをコードするｄｎａ | |
| CA2354030A1 (en) | Micromonospora echinospora genes encoding for biosynthesis of calicheamicin and self-resistance thereto | |
| CN102021187A (zh) | Fr-008聚酮抗生素生物合成相关的基因簇 | |
| WO2006053313A2 (en) | Elaiophylin biosynthetic gene cluster | |
| JP2009502187A (ja) | チオコラリンの生合成に関与する遺伝子およびその異種産生 | |
| HK1187956A (zh) | 鑒定和表徵來自生產多殺菌素的刺糖多孢菌的siponactin生物合成基因簇 |