JP4245477B2 - 大環状ポリケチドを特定するための方法 - Google Patents
大環状ポリケチドを特定するための方法 Download PDFInfo
- Publication number
- JP4245477B2 JP4245477B2 JP2003519515A JP2003519515A JP4245477B2 JP 4245477 B2 JP4245477 B2 JP 4245477B2 JP 2003519515 A JP2003519515 A JP 2003519515A JP 2003519515 A JP2003519515 A JP 2003519515A JP 4245477 B2 JP4245477 B2 JP 4245477B2
- Authority
- JP
- Japan
- Prior art keywords
- gene
- coli
- host cell
- mphr
- mph
- 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
- 229930001119 polyketide Natural products 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 31
- 125000000830 polyketide group Chemical group 0.000 title claims abstract description 26
- 108090000623 proteins and genes Proteins 0.000 claims description 43
- 241000588724 Escherichia coli Species 0.000 claims description 31
- 238000003556 assay Methods 0.000 claims description 16
- 108700008625 Reporter Genes Proteins 0.000 claims description 15
- 150000003881 polyketide derivatives Chemical class 0.000 claims description 15
- 108700039691 Genetic Promoter Regions Proteins 0.000 claims description 13
- 108060001084 Luciferase Proteins 0.000 claims description 12
- 239000005089 Luciferase Substances 0.000 claims description 11
- 239000003120 macrolide antibiotic agent Substances 0.000 claims description 11
- 102000004169 proteins and genes Human genes 0.000 claims description 9
- 108010043121 Green Fluorescent Proteins Proteins 0.000 claims description 7
- 102000004144 Green Fluorescent Proteins Human genes 0.000 claims description 7
- 239000005090 green fluorescent protein Substances 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 6
- 230000035772 mutation Effects 0.000 claims description 6
- 238000013518 transcription Methods 0.000 claims description 6
- 230000035897 transcription Effects 0.000 claims description 6
- 101150071242 tolC gene Proteins 0.000 claims description 5
- 241000894006 Bacteria Species 0.000 claims description 3
- 102000005421 acetyltransferase Human genes 0.000 claims description 3
- 108020002494 acetyltransferase Proteins 0.000 claims description 3
- 108010005774 beta-Galactosidase Proteins 0.000 claims description 3
- 102000005936 beta-Galactosidase Human genes 0.000 claims description 3
- 238000003571 reporter gene assay Methods 0.000 claims description 3
- 230000002018 overexpression Effects 0.000 claims description 2
- 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 24
- 239000013612 plasmid Substances 0.000 description 19
- 229920001817 Agar Polymers 0.000 description 17
- 239000008272 agar Substances 0.000 description 17
- 238000004020 luminiscence type Methods 0.000 description 15
- 239000000126 substance Substances 0.000 description 15
- 210000004027 cell Anatomy 0.000 description 13
- 108020004414 DNA Proteins 0.000 description 12
- 229960003276 erythromycin Drugs 0.000 description 12
- 238000009792 diffusion process Methods 0.000 description 9
- 230000027455 binding Effects 0.000 description 8
- 238000001514 detection method Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 239000012634 fragment Substances 0.000 description 6
- 150000002596 lactones Chemical class 0.000 description 6
- 239000004098 Tetracycline Substances 0.000 description 5
- 230000014509 gene expression Effects 0.000 description 5
- 229940041033 macrolides Drugs 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- 235000019364 tetracycline Nutrition 0.000 description 5
- 230000009471 action Effects 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 229930014626 natural product Natural products 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 229960002180 tetracycline Drugs 0.000 description 4
- 229930101283 tetracycline Natural products 0.000 description 4
- 150000003522 tetracyclines Chemical class 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 238000011065 in-situ storage Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 3
- 229960001225 rifampicin Drugs 0.000 description 3
- HJYYPODYNSCCOU-ODRIEIDWSA-N rifamycin SV Chemical compound OC1=C(C(O)=C2C)C3=C(O)C=C1NC(=O)\C(C)=C/C=C/[C@H](C)[C@H](O)[C@@H](C)[C@@H](O)[C@@H](C)[C@H](OC(C)=O)[C@H](C)[C@@H](OC)\C=C\O[C@@]1(C)OC2=C3C1=O HJYYPODYNSCCOU-ODRIEIDWSA-N 0.000 description 3
- 230000007017 scission Effects 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 230000002103 transcriptional effect Effects 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- 241000607620 Aliivibrio fischeri Species 0.000 description 2
- 206010010144 Completed suicide Diseases 0.000 description 2
- 238000007399 DNA isolation Methods 0.000 description 2
- 206010017533 Fungal infection Diseases 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- RZPAKFUAFGMUPI-UHFFFAOYSA-N Oleandomycin Natural products O1C(C)C(O)C(OC)CC1OC1C(C)C(=O)OC(C)C(C)C(O)C(C)C(=O)C2(OC2)CC(C)C(OC2C(C(CC(C)O2)N(C)C)O)C1C RZPAKFUAFGMUPI-UHFFFAOYSA-N 0.000 description 2
- 239000004104 Oleandomycin Substances 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 229940123573 Protein synthesis inhibitor Drugs 0.000 description 2
- 241000187559 Saccharopolyspora erythraea Species 0.000 description 2
- 108010059993 Vancomycin Proteins 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 230000000843 anti-fungal effect Effects 0.000 description 2
- -1 aromatic polyketide tetracyclines Chemical class 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 239000003623 enhancer Substances 0.000 description 2
- 229930013356 epothilone Natural products 0.000 description 2
- 150000003883 epothilone derivatives Chemical class 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 229960002351 oleandomycin Drugs 0.000 description 2
- RZPAKFUAFGMUPI-KGIGTXTPSA-N oleandomycin Chemical compound O1[C@@H](C)[C@H](O)[C@@H](OC)C[C@@H]1O[C@@H]1[C@@H](C)C(=O)O[C@H](C)[C@H](C)[C@H](O)[C@@H](C)C(=O)[C@]2(OC2)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 RZPAKFUAFGMUPI-KGIGTXTPSA-N 0.000 description 2
- 235000019367 oleandomycin Nutrition 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000007 protein synthesis inhibitor Substances 0.000 description 2
- 238000007423 screening assay Methods 0.000 description 2
- 230000005945 translocation Effects 0.000 description 2
- 229960003165 vancomycin Drugs 0.000 description 2
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 2
- MYPYJXKWCTUITO-LYRMYLQWSA-O vancomycin(1+) Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C([O-])=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)[NH2+]C)[C@H]1C[C@](C)([NH3+])[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-O 0.000 description 2
- 239000005660 Abamectin Substances 0.000 description 1
- 101710201211 Acyl-[acyl-carrier-protein]-UDP-N-acetylglucosamine O-acyltransferase Proteins 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 229930188120 Carbomycin Natural products 0.000 description 1
- 229940123982 Cell wall synthesis inhibitor Drugs 0.000 description 1
- 108010069514 Cyclic Peptides Proteins 0.000 description 1
- 102000001189 Cyclic Peptides Human genes 0.000 description 1
- 230000004568 DNA-binding Effects 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000331 Firefly luciferases Proteins 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- IEMDOFXTVAPVLX-YWQHLDGFSA-N Leucomycin A1 Chemical compound CO[C@H]1[C@H](O)CC(=O)O[C@H](C)C\C=C\C=C\[C@H](O)[C@H](C)C[C@H](CC=O)[C@@H]1O[C@H]1[C@H](O)[C@@H](N(C)C)[C@H](O[C@@H]2O[C@@H](C)[C@H](OC(=O)CC(C)C)[C@](C)(O)C2)[C@@H](C)O1 IEMDOFXTVAPVLX-YWQHLDGFSA-N 0.000 description 1
- 239000006142 Luria-Bertani Agar Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000031888 Mycoses Diseases 0.000 description 1
- RMIXHJPMNBXMBU-QIIXEHPYSA-N Nonactin Chemical compound C[C@H]([C@H]1CC[C@H](O1)C[C@@H](OC(=O)[C@@H](C)[C@@H]1CC[C@@H](O1)C[C@@H](C)OC(=O)[C@H](C)[C@H]1CC[C@H](O1)C[C@H](C)OC(=O)[C@H]1C)C)C(=O)O[C@H](C)C[C@H]2CC[C@@H]1O2 RMIXHJPMNBXMBU-QIIXEHPYSA-N 0.000 description 1
- RMIXHJPMNBXMBU-UHFFFAOYSA-N Nonactin Natural products CC1C(=O)OC(C)CC(O2)CCC2C(C)C(=O)OC(C)CC(O2)CCC2C(C)C(=O)OC(C)CC(O2)CCC2C(C)C(=O)OC(C)CC2CCC1O2 RMIXHJPMNBXMBU-UHFFFAOYSA-N 0.000 description 1
- YJQPYGGHQPGBLI-UHFFFAOYSA-N Novobiocin Natural products O1C(C)(C)C(OC)C(OC(N)=O)C(O)C1OC1=CC=C(C(O)=C(NC(=O)C=2C=C(CC=C(C)C)C(O)=CC=2)C(=O)O2)C2=C1C YJQPYGGHQPGBLI-UHFFFAOYSA-N 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 101710190786 PI protein Proteins 0.000 description 1
- UZQBOFAUUTZOQE-UHFFFAOYSA-N Pikromycin Natural products CC1CC(C)C(=O)C=CC(O)(C)C(CC)OC(=O)C(C)C(=O)C(C)C1OC1C(O)C(N(C)C)CC(C)O1 UZQBOFAUUTZOQE-UHFFFAOYSA-N 0.000 description 1
- 229930189077 Rifamycin Natural products 0.000 description 1
- 241000634742 Saccharopolyspora erythraea NRRL 2338 Species 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- FQVHOULQCKDUCY-OGHXVOSASA-N [(2s,3s,4r,6s)-6-[(2r,3s,4r,5r,6s)-6-[[(1s,3r,7r,8s,9s,10r,12r,14e,16s)-7-acetyloxy-8-methoxy-3,12-dimethyl-5,13-dioxo-10-(2-oxoethyl)-4,17-dioxabicyclo[14.1.0]heptadec-14-en-9-yl]oxy]-4-(dimethylamino)-5-hydroxy-2-methyloxan-3-yl]oxy-4-hydroxy-2,4-dimeth 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](CC=O)C[C@@H](C)C(=O)/C=C/[C@@H]2O[C@H]2C[C@@H](C)OC(=O)C[C@H]([C@@H]1OC)OC(C)=O)[C@H]1C[C@@](C)(O)[C@@H](OC(=O)CC(C)C)[C@H](C)O1 FQVHOULQCKDUCY-OGHXVOSASA-N 0.000 description 1
- 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 1
- 230000004913 activation Effects 0.000 description 1
- 239000000362 adenosine triphosphatase inhibitor Substances 0.000 description 1
- 108700029371 albomycin Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 229930014544 aromatic polyketide Natural products 0.000 description 1
- RRZXIRBKKLTSOM-XPNPUAGNSA-N avermectin B1a Chemical compound C1=C[C@H](C)[C@@H]([C@@H](C)CC)O[C@]11O[C@H](C\C=C(C)\[C@@H](O[C@@H]2O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](OC)C3)[C@@H](OC)C2)[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 RRZXIRBKKLTSOM-XPNPUAGNSA-N 0.000 description 1
- 229960004099 azithromycin Drugs 0.000 description 1
- MQTOSJVFKKJCRP-BICOPXKESA-N azithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)N(C)C[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 MQTOSJVFKKJCRP-BICOPXKESA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229950005779 carbomycin Drugs 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- ZCDOYSPFYFSLEW-UHFFFAOYSA-N chromate(2-) Chemical class [O-][Cr]([O-])(=O)=O ZCDOYSPFYFSLEW-UHFFFAOYSA-N 0.000 description 1
- 229960002626 clarithromycin Drugs 0.000 description 1
- AGOYDEPGAOXOCK-KCBOHYOISA-N clarithromycin 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)([C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)OC)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 AGOYDEPGAOXOCK-KCBOHYOISA-N 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000001085 cytostatic effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000013070 direct material Substances 0.000 description 1
- 244000078703 ectoparasite Species 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 244000079386 endoparasite Species 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229960000308 fosfomycin Drugs 0.000 description 1
- YMDXZJFXQJVXBF-STHAYSLISA-N fosfomycin Chemical compound C[C@@H]1O[C@@H]1P(O)(O)=O YMDXZJFXQJVXBF-STHAYSLISA-N 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 150000002337 glycosamines Chemical class 0.000 description 1
- 239000002271 gyrase inhibitor Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 229940121354 immunomodulator Drugs 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 239000002917 insecticide Substances 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 150000003951 lactams Chemical class 0.000 description 1
- GZQKNULLWNGMCW-PWQABINMSA-N lipid A (E. coli) Chemical compound O1[C@H](CO)[C@@H](OP(O)(O)=O)[C@H](OC(=O)C[C@@H](CCCCCCCCCCC)OC(=O)CCCCCCCCCCCCC)[C@@H](NC(=O)C[C@@H](CCCCCCCCCCC)OC(=O)CCCCCCCCCCC)[C@@H]1OC[C@@H]1[C@@H](O)[C@H](OC(=O)C[C@H](O)CCCCCCCCCCC)[C@@H](NC(=O)C[C@H](O)CCCCCCCCCCC)[C@@H](OP(O)(O)=O)O1 GZQKNULLWNGMCW-PWQABINMSA-N 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- HUKYPYXOBINMND-UHFFFAOYSA-N methymycin Natural products CC1CC(C)C(=O)C=CC(O)(C)C(CC)OC(=O)C(C)C1OC1C(O)C(N(C)C)CC(C)O1 HUKYPYXOBINMND-UHFFFAOYSA-N 0.000 description 1
- 238000013048 microbiological method Methods 0.000 description 1
- FXWHFKOXMBTCMP-WMEDONTMSA-N milbemycin Natural products COC1C2OCC3=C/C=C/C(C)CC(=CCC4CC(CC5(O4)OC(C)C(C)C(OC(=O)C(C)CC(C)C)C5O)OC(=O)C(C=C1C)C23O)C FXWHFKOXMBTCMP-WMEDONTMSA-N 0.000 description 1
- 229960002950 novobiocin Drugs 0.000 description 1
- YJQPYGGHQPGBLI-KGSXXDOSSA-N novobiocin Chemical compound O1C(C)(C)[C@H](OC)[C@@H](OC(N)=O)[C@@H](O)[C@@H]1OC1=CC=C(C(O)=C(NC(=O)C=2C=C(CC=C(C)C)C(O)=CC=2)C(=O)O2)C2=C1C YJQPYGGHQPGBLI-KGSXXDOSSA-N 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 229960000988 nystatin Drugs 0.000 description 1
- VQOXZBDYSJBXMA-NQTDYLQESA-N nystatin A1 Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/CC/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 VQOXZBDYSJBXMA-NQTDYLQESA-N 0.000 description 1
- UZQBOFAUUTZOQE-VSLWXVDYSA-N pikromycin Chemical compound C[C@H]1C[C@@H](C)C(=O)\C=C\[C@@](O)(C)[C@@H](CC)OC(=O)[C@H](C)C(=O)[C@H](C)[C@H]1O[C@H]1[C@H](O)[C@@H](N(C)C)C[C@@H](C)O1 UZQBOFAUUTZOQE-VSLWXVDYSA-N 0.000 description 1
- 150000004291 polyenes Chemical class 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- ZAHRKKWIAAJSAO-UHFFFAOYSA-N rapamycin Natural products COCC(O)C(=C/C(C)C(=O)CC(OC(=O)C1CCCCN1C(=O)C(=O)C2(O)OC(CC(OC)C(=CC=CC=CC(C)CC(C)C(=O)C)C)CCC2C)C(C)CC3CCC(O)C(C3)OC)C ZAHRKKWIAAJSAO-UHFFFAOYSA-N 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 229960003292 rifamycin Drugs 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- QFJCIRLUMZQUOT-HPLJOQBZSA-N sirolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 QFJCIRLUMZQUOT-HPLJOQBZSA-N 0.000 description 1
- 229960002930 sirolimus Drugs 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 229960001967 tacrolimus Drugs 0.000 description 1
- 229940040944 tetracyclines Drugs 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 230000017105 transposition Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6897—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids involving reporter genes operably linked to promoters
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
- G01N33/9446—Antibacterials
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Urology & Nephrology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Hematology (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmacology & Pharmacy (AREA)
- Cell Biology (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
プラスミドpEBZ511の作製
最初に、PCR増幅を使用して、mph(A)オペロンオペロンのプロモーター及びMphRタンパク質が結合するDNA配列を得ることにより、プラスミドpEBZ511を作製した。使用した鋳型は、プラスミドpTZ3509(Noguchiら、2000)のDNAであった。使用したプライマーは、末端BamHI及びEcoRI認識配列を有する所望のDNAフラグメントを増幅するのに役立つオリゴヌクレオチドmphf216及びmphr217であった。PCR混合物を最初に95℃で3分間変性し、その後95℃で60秒間の変性、58℃で90秒間のアニーリング及び68℃で2分間の伸長の30サイクルに供した。このPCR混合物は、前記の2個のプライマー100pmol、pTZ3509 10ng、dNTP 0.5mM、1×Pfx緩衝液(Life Technologies)、1×PCRエンハンサー溶液(Life Technologies)及びPfxポリメラーゼ5U(Life Technologies)を含んだ。標準分子生物学法(Sambrookら、1989)を用いて、得られたDNAアンプリコンを単離し、BamHI及びEcoRIで切断して、BamHIとEcoRIで切断しておいたベクターpUCD615(Rogowskyら、1987)にクローニングした。
mph(A)オペロンを担う自殺トランスポゾンプラスミドpEBZ512の作製及び大腸菌SM101へのmph(A)オペロンの転位
PstIを使用して、プラスミドpTZ3509(Noguchiら、2000)から4.1kb DNAフラグメント上でmph(A)オペロンを切り出し、標準分子生物学法を用いて、自殺トランスポゾンベクターpBSL180(AlexeyevとShokolenko、1995)のPstI開裂部位、プラスミドの転移性部分にクローニングした。形質転換のために使用した宿主は、大腸菌CC118 Lambdapir(Herreroら、1990)であった。この菌株は、プラスミドの複製に必要なπタンパク質を有する。作製したプラスミドをpEBZ512と称した。標準形質転換法を用いてこのプラスミドを大腸菌SM101に導入し、mph(A)オペロンを担うトランスポゾンの転位後に形質転換体を選択した。この手順によってゲノムに組み込まれたDNAのmph(A)オペロンを担う転位断片をEBZ512と称した。標準的方法(Sambrookら、1989)を用いて分子生物学的作業(DNA単離、制限、連結及び形質転換)を実施した。
mphR(A)をコードするプラスミドpEBZ514の作製
プライマーMLV637及びMLV638及びpEBZ512−DNAを用いて、PCRによってmphR(A)の遺伝子領域を増幅した。リボソーム結合部位及びSphI開裂部位をプライマーMLV637に組み込み、SalI開裂部位をプライマーMLV638に組み込んだ。Mastercycler Gradient型エッペンドルフサーモサイクラーでPCR反応を実施した。PCR混合物を最初に96℃で10分間変性し、その後96℃で60秒間の変性、56℃で90秒間のアニーリング及び72℃で40秒間の伸長という30回の伸長サイクルに供した。このPCR混合物は、前記の2個のプライマー100pmol、pEBZ512−DNA 10ng、dNTP 0.5mM、1×Pfx緩衝液(Life Technologies)、1×PCRエンハンサー溶液(Life Technologies)及びPfxポリメラーゼ5U(Life Technologies)を含んだ。生成されたDNAフラグメントを単離し、制限酵素SphI及びSalIで切断して、SphIとSalIで同様に切断しておいたベクターpACYC184(ChangとCohen、1978)に連結した。mphR(A)をコードする領域は、従って、対応する耐性遺伝子プロモーターの下流の、テトラサイクリン耐性遺伝子内に位置した。標準的方法(Sambrookら、1989)を用いて分子生物学的作業(DNA単離、制限、連結及び形質転換)を実施した。
バイオセンサー、大腸菌SM101(EBZ512、pEBZ511、pEBZ514)をLB培地で対数増殖後期まで好気的に増殖させ、標準微生物学的方法を用いて軟寒天に混合して、クロラムフェニコール(25μg/ml)及びカナマイシン(30μg/ml)を含むLB−寒天平板に塗布した。軟寒天が凝固した後、検定する物質を既に含む感受性アッセイディスク又は検定する物質を適用されている非充填アッセイディスク(Oxoid GmbH,Am Lippeglacis 4−8,46483 Wesel,Germany)を寒天上に置いた。この寒天平板拡散アッセイを28℃で2時間から4時間又は15時間から24時間インキュベートし、Berthold Luminograph LB980を用いて発光を記録した(図3、4及び5)。発光が促進される場合は、配置した試料の周囲の拡散帯域内で検出できる。検定する試料が、同時に、より長時間のインキュベーションの非常に好ましい実施形態において阻害帯域として示される、センサー生物に対する抗菌活性を有する場合は、発光の促進は、致死量以下の濃度範囲内でこの阻害の輪の外側で検出されうる。
バイオセンサーを使用して、大環状ポリケチドの生産生物として知られる微生物をもう1つ別の微生物からインサイチューで識別することも可能であった。このために、微生物バイオセンサー細胞を寒天平板拡散アッセイにおいて使用した。寒天ブロックを、軟寒天に包埋したバイオセンサー懸濁液上に置いた。これらの寒天ブロックを、エリスロマイシン生産生物として記述されているサッカロポリスポラ・エリトラエア(Saccharopolyspora erythraea)DSM40517又はアビラマイシン生産生物として記述されているストレプトマイセス・ビリドクロモゲネス(Streptomyces viridochromogenes)DSM40721のいずれかで被覆した。もう1つの寒天ブロックは増殖物を含まなかった。発光の促進は、付加的に適用したエリスロマイシンアッセイディスク及びサッカロポリスポラ・エリトラエア被覆寒天ブロックに関してのみ検出可能であった(図5)。
ブタペスト条約の必要条件に従って、2001年6月6日に、下記の菌株をthe Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH(DSMZ),Mascheroder Weg 1b,D−38124 Brunswick,Germanyに寄託した:
Claims (17)
- 大環状ポリケチド又は大環状ポリケチドを含む混合物を細胞レポーター−遺伝子アッセイ系に供することを特徴とする、大環状ポリケチドを特定するための方法であって、
前記アッセイ系は、転写が大腸菌mph(A)遺伝子のプロモーター領域の制御下にあるレポーター遺伝子を含み、且つ、大腸菌MphR(A)タンパク質を含み、前記MphR(A)タンパク質は、mphR(A)遺伝子を過剰発現することによって提供される前記方法。 - 前記大環状ポリケチドがマクロライドであることを特徴とする、請求項1に記載の方法。
- 使用する前記レポーター遺伝子が、クロラムフェニルコールアセチルトランスフェラーゼ、β−ガラクトシダーゼ、ルシフェラーゼ又はグリーン蛍光タンパク質(GFP)をコードする遺伝子であることを特徴とする、請求項1または2に記載の方法。
- 使用する前記レポーター遺伝子が、ルシフェラーゼをコードする遺伝子であることを特徴とする、請求項3に記載の方法。
- 前記アッセイ系が、完全な大腸菌mph(A)オペロン又はその部分をさらに含むことを特徴とする、請求項1から4のいずれかに記載の方法。
- 前記細菌が大腸菌であることを特徴とする、請求項1から5のいずれかに記載の方法。
- 前記大腸菌が、DSM14334の下に寄託されている大腸菌SM101(EBZ512、pEBZ511、pEBZ514)であることを特徴とする、請求項6に記載の方法。
- 前記大腸菌が、lpxA2遺伝子及び/又はtolC遺伝子内に突然変異を有する菌株であることを特徴とする、請求項6に記載の方法。
- 転写が大腸菌mph(A)遺伝子のプロモーター領域の制御下にあるレポーター遺伝子、および大腸菌MphR(A)タンパク質を含む宿主細胞であって、前記MphR(A)タンパク質は、mphR(A)遺伝子を過剰発現することによって提供される前記宿主細胞。
- 前記レポーター遺伝子が、クロラムフェニルコールアセチルトランスフェラーゼ、β−ガラクトシダーゼ、ルシフェラーゼ又はグリーン蛍光タンパク質(GFP)をコードする遺伝子であることを特徴とする、請求項9に記載の宿主細胞。
- 前記レポーター遺伝子がルシフェラーゼをコードする遺伝子であることを特徴とする、請求項10に記載の宿主細胞。
- 完全な大腸菌mph(A)オペロンを含むことを特徴とする、請求項9から11のいずれかに記載の宿主細胞。
- 大腸菌であることを特徴とする、請求項9から12のいずれかに記載の宿主細胞。
- DSM14334の下に寄託されている大腸菌SM101(EBZ512、pEBZ511、pEBZ514)であることを特徴とする、請求項13に記載の宿主細胞。
- 前記大腸菌が、lpxA2遺伝子及び/又はtolC遺伝子内に突然変異を有する菌株であることを特徴とする、請求項13に記載の宿主細胞。
- 大環状ポリケチドを特定するための、請求項9から15のいずれかに記載の宿主細胞の使用。
- 請求項9から15のいずれかに記載の宿主細胞を含むキット。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE10138766A DE10138766A1 (de) | 2001-08-07 | 2001-08-07 | Verfahren zum Identifizieren von makrocyclischen Polyketiden |
PCT/EP2002/008282 WO2003014386A2 (de) | 2001-08-07 | 2002-07-25 | Verfahren zum identifizieren von makrocyclischen polyketiden |
Publications (3)
Publication Number | Publication Date |
---|---|
JP2004538007A JP2004538007A (ja) | 2004-12-24 |
JP2004538007A5 JP2004538007A5 (ja) | 2006-01-05 |
JP4245477B2 true JP4245477B2 (ja) | 2009-03-25 |
Family
ID=7694679
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2003519515A Expired - Fee Related JP4245477B2 (ja) | 2001-08-07 | 2002-07-25 | 大環状ポリケチドを特定するための方法 |
Country Status (7)
Country | Link |
---|---|
US (1) | US20040209270A1 (ja) |
EP (1) | EP1417346B1 (ja) |
JP (1) | JP4245477B2 (ja) |
AT (1) | ATE415492T1 (ja) |
DE (2) | DE10138766A1 (ja) |
ES (1) | ES2315389T3 (ja) |
WO (1) | WO2003014386A2 (ja) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6336504B1 (en) | 2000-03-03 | 2002-01-08 | Pancanadian Petroleum Limited | Downhole separation and injection of produced water in naturally flowing or gas-lifted hydrocarbon wells |
US6336503B1 (en) | 2000-03-03 | 2002-01-08 | Pancanadian Petroleum Limited | Downhole separation of produced water in hydrocarbon wells, and simultaneous downhole injection of separated water and surface water |
WO2012045451A1 (en) | 2010-10-05 | 2012-04-12 | Ludwig-Maximilians-Universitaet Muenchen | Novel therapeutic treatment of progranulin-dependent diseases |
WO2017196983A1 (en) * | 2016-05-10 | 2017-11-16 | North Carolina State University | Genetically encoded biosensors for detection of polyketides |
US20220243288A1 (en) * | 2019-05-21 | 2022-08-04 | North Carolina State University | Biosensors for polyketide extender units and uses thereof |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ATE165394T1 (de) * | 1991-03-01 | 1998-05-15 | Vito | Fusionierte gene sowie deren verwendungzur bestimmung von metall- und xenobiotikagehalten |
IL107815A (en) * | 1992-12-04 | 2003-12-10 | Du Pont | Genetic constructs comprising a stress-responsive promoter linked to a lux reporter operon and methods of use in environmental testing |
US6242211B1 (en) * | 1996-04-24 | 2001-06-05 | Terragen Discovery, Inc. | Methods for generating and screening novel metabolic pathways |
US5976813A (en) * | 1997-12-12 | 1999-11-02 | Abbott Laboratories | Continuous format high throughput screening |
-
2001
- 2001-08-07 DE DE10138766A patent/DE10138766A1/de not_active Withdrawn
-
2002
- 2002-07-25 EP EP02760269A patent/EP1417346B1/de not_active Expired - Lifetime
- 2002-07-25 WO PCT/EP2002/008282 patent/WO2003014386A2/de active Application Filing
- 2002-07-25 JP JP2003519515A patent/JP4245477B2/ja not_active Expired - Fee Related
- 2002-07-25 DE DE50213063T patent/DE50213063D1/de not_active Expired - Fee Related
- 2002-07-25 AT AT02760269T patent/ATE415492T1/de not_active IP Right Cessation
- 2002-07-25 ES ES02760269T patent/ES2315389T3/es not_active Expired - Lifetime
- 2002-07-25 US US10/486,187 patent/US20040209270A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
US20040209270A1 (en) | 2004-10-21 |
WO2003014386A2 (de) | 2003-02-20 |
DE50213063D1 (de) | 2009-01-08 |
DE10138766A1 (de) | 2003-02-20 |
JP2004538007A (ja) | 2004-12-24 |
ES2315389T3 (es) | 2009-04-01 |
ATE415492T1 (de) | 2008-12-15 |
EP1417346A2 (de) | 2004-05-12 |
EP1417346B1 (de) | 2008-11-26 |
WO2003014386A3 (de) | 2004-01-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Cao et al. | Antibiotics that inhibit cell wall biosynthesis induce expression of the Bacillus subtilisσW and σM regulons | |
Schweizer | Vectors to express foreign genes and techniques to monitor gene expression in Pseudomonads | |
EP0922106B1 (en) | Light emitting biosensor | |
Serizawa et al. | Functional analysis of the YvrGHb two-component system of Bacillus subtilis: identification of the regulated genes by DNA microarray and northern blot analyses | |
JP4245477B2 (ja) | 大環状ポリケチドを特定するための方法 | |
Lu et al. | LysR family transcriptional regulator PqsR as repressor of pyoluteorin biosynthesis and activator of phenazine-1-carboxylic acid biosynthesis in Pseudomonas sp. M18 | |
JP3329395B2 (ja) | テトラサイクリンのスクリーニング法 | |
He et al. | SCO3129, a TetR family regulator, is responsible for osmotic stress in Streptomyces coelicolor | |
US8389263B2 (en) | Spores for the stabilization and on-site application of bacterial whole-cell biosensing systems | |
Bachmann et al. | High-throughput identification and validation of in situ-expressed genes of Lactococcus lactis | |
WO2004106557A2 (en) | Thiamin production by fermentation | |
CA2452849C (en) | Method for monitoring and modulating protein folding | |
KR101292375B1 (ko) | 톡소플라빈 검출용 바이오센스 균주 및 이를 이용한 톡소플라빈 검출방법 | |
Kotova et al. | Inducible specific lux-biosensors for the detection of antibiotics: Construction and main parameters | |
Park et al. | Characterization of quorum-sensing signaling molecules produced by Burkholderia cepacia G4 | |
JP2004248596A (ja) | 抗菌剤のスクリーニング方法およびそれに用いる微生物 | |
KR100997044B1 (ko) | 시그마 인자 r 및 이를 이용한 항생제 생산 증가 방법 | |
US20060035365A1 (en) | Method for identifying cellular growth inhibitors | |
AU722016B2 (en) | Biosensors | |
US7229814B2 (en) | Nucleic acids and polypeptides involved in the production of cryptophycin | |
US20080050759A1 (en) | Screening Assay for Anti-Bacterial Compounds | |
US20040241708A1 (en) | Recombinant vector for transforming strain to detect benzoic acid and derivatives thereof, transformant containing the recombinant vector, and method for detecting benzoic acid and derivatives thereof using the transformant | |
EP1608779A1 (en) | Topoisomerase modulator assays |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20050606 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20050606 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20080812 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20081111 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20081209 |
|
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20090106 |
|
R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120116 Year of fee payment: 3 |
|
LAPS | Cancellation because of no payment of annual fees |