NO301487B1 - Isolert nukleinsyremolekyl kodende for en termisk stabil cytosin deaminase, rekombinant ekspresjonsvektor, vertscelle og fremgangsmåte for fremstilling av rekombinant termisk stabil cytosin deaminase - Google Patents
Isolert nukleinsyremolekyl kodende for en termisk stabil cytosin deaminase, rekombinant ekspresjonsvektor, vertscelle og fremgangsmåte for fremstilling av rekombinant termisk stabil cytosin deaminase Download PDFInfo
- Publication number
- NO301487B1 NO301487B1 NO902551A NO902551A NO301487B1 NO 301487 B1 NO301487 B1 NO 301487B1 NO 902551 A NO902551 A NO 902551A NO 902551 A NO902551 A NO 902551A NO 301487 B1 NO301487 B1 NO 301487B1
- Authority
- NO
- Norway
- Prior art keywords
- cdase
- thermally stable
- cytosine deaminase
- sequence
- yeast
- Prior art date
Links
- 102000000311 Cytosine Deaminase Human genes 0.000 title claims abstract description 25
- 108010080611 Cytosine Deaminase Proteins 0.000 title claims abstract description 25
- 239000013604 expression vector Substances 0.000 title claims description 16
- 108020004707 nucleic acids Proteins 0.000 title claims description 11
- 102000039446 nucleic acids Human genes 0.000 title claims description 11
- 150000007523 nucleic acids Chemical class 0.000 title claims description 11
- 238000004519 manufacturing process Methods 0.000 title claims description 8
- 238000003259 recombinant expression Methods 0.000 title claims description 8
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 83
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 57
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 51
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 50
- 102000004190 Enzymes Human genes 0.000 claims abstract description 43
- 108090000790 Enzymes Proteins 0.000 claims abstract description 43
- 238000000034 method Methods 0.000 claims abstract description 31
- 238000001641 gel filtration chromatography Methods 0.000 claims abstract description 3
- 238000006243 chemical reaction Methods 0.000 claims description 40
- XRECTZIEBJDKEO-UHFFFAOYSA-N flucytosine Chemical compound NC1=NC(=O)NC=C1F XRECTZIEBJDKEO-UHFFFAOYSA-N 0.000 claims description 31
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 29
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 claims description 25
- 229960002949 fluorouracil Drugs 0.000 claims description 25
- 230000014509 gene expression Effects 0.000 claims description 24
- 229960004413 flucytosine Drugs 0.000 claims description 20
- 238000000746 purification Methods 0.000 claims description 19
- 238000002955 isolation Methods 0.000 claims description 7
- 230000004071 biological effect Effects 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 4
- 108010032220 cyclomaltodextrinase Proteins 0.000 description 152
- 210000004027 cell Anatomy 0.000 description 41
- 230000000694 effects Effects 0.000 description 34
- 238000003752 polymerase chain reaction Methods 0.000 description 34
- 239000000872 buffer Substances 0.000 description 26
- 239000012634 fragment Substances 0.000 description 25
- 108020004414 DNA Proteins 0.000 description 24
- 239000013615 primer Substances 0.000 description 23
- 108091026890 Coding region Proteins 0.000 description 22
- 239000013598 vector Substances 0.000 description 20
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 19
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 19
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 19
- 235000011130 ammonium sulphate Nutrition 0.000 description 19
- 239000002773 nucleotide Substances 0.000 description 19
- 125000003729 nucleotide group Chemical group 0.000 description 19
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 18
- 239000002953 phosphate buffered saline Substances 0.000 description 18
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 17
- 239000000499 gel Substances 0.000 description 17
- 239000000243 solution Substances 0.000 description 15
- 238000010367 cloning Methods 0.000 description 14
- 108090000765 processed proteins & peptides Proteins 0.000 description 14
- 108091028043 Nucleic acid sequence Proteins 0.000 description 13
- 102000004196 processed proteins & peptides Human genes 0.000 description 13
- 150000001413 amino acids Chemical class 0.000 description 12
- 239000013612 plasmid Substances 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 238000004458 analytical method Methods 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 238000012163 sequencing technique Methods 0.000 description 11
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 239000002609 medium Substances 0.000 description 10
- 229920001184 polypeptide Polymers 0.000 description 10
- 239000000523 sample Substances 0.000 description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 108091034117 Oligonucleotide Proteins 0.000 description 9
- ATDGTVJJHBUTRL-UHFFFAOYSA-N cyanogen bromide Chemical compound BrC#N ATDGTVJJHBUTRL-UHFFFAOYSA-N 0.000 description 9
- 230000004927 fusion Effects 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- 206010028980 Neoplasm Diseases 0.000 description 8
- 108010076504 Protein Sorting Signals Proteins 0.000 description 8
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 8
- 239000007983 Tris buffer Substances 0.000 description 8
- 230000036961 partial effect Effects 0.000 description 8
- 239000000758 substrate Substances 0.000 description 8
- 238000001890 transfection Methods 0.000 description 8
- 241000894006 Bacteria Species 0.000 description 7
- 238000012300 Sequence Analysis Methods 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
- QTBSBXVTEAMEQO-UHFFFAOYSA-N acetic acid Substances CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 7
- 238000003776 cleavage reaction Methods 0.000 description 7
- 239000002299 complementary DNA Substances 0.000 description 7
- 238000009396 hybridization Methods 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 239000002244 precipitate Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- 230000001105 regulatory effect Effects 0.000 description 7
- 108091008146 restriction endonucleases Proteins 0.000 description 7
- 230000007017 scission Effects 0.000 description 7
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 6
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 229920005654 Sephadex Polymers 0.000 description 6
- 239000012507 Sephadex™ Substances 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000007792 addition Methods 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 6
- 238000010828 elution Methods 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 238000000108 ultra-filtration Methods 0.000 description 6
- KFDVPJUYSDEJTH-UHFFFAOYSA-N 4-ethenylpyridine Chemical compound C=CC1=CC=NC=C1 KFDVPJUYSDEJTH-UHFFFAOYSA-N 0.000 description 5
- 108010051815 Glutamyl endopeptidase Proteins 0.000 description 5
- 108010030544 Peptidyl-Lys metalloendopeptidase Proteins 0.000 description 5
- 238000010276 construction Methods 0.000 description 5
- 229930182830 galactose Natural products 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 210000004962 mammalian cell Anatomy 0.000 description 5
- 230000035772 mutation Effects 0.000 description 5
- 239000008057 potassium phosphate buffer Substances 0.000 description 5
- 238000004007 reversed phase HPLC Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 108020004705 Codon Proteins 0.000 description 4
- 229920002307 Dextran Polymers 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- 241000235070 Saccharomyces Species 0.000 description 4
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 4
- 239000002246 antineoplastic agent Substances 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 108010003914 endoproteinase Asp-N Proteins 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 239000012091 fetal bovine serum Substances 0.000 description 4
- 108020001507 fusion proteins Proteins 0.000 description 4
- 102000037865 fusion proteins Human genes 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 238000003780 insertion Methods 0.000 description 4
- 230000037431 insertion Effects 0.000 description 4
- 239000006166 lysate Substances 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 108091033319 polynucleotide Proteins 0.000 description 4
- 102000040430 polynucleotide Human genes 0.000 description 4
- 239000002157 polynucleotide Substances 0.000 description 4
- -1 polypropylene Polymers 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 238000006467 substitution reaction Methods 0.000 description 4
- 238000013518 transcription Methods 0.000 description 4
- 230000035897 transcription Effects 0.000 description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 4
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 208000035404 Autolysis Diseases 0.000 description 3
- 101150041795 CDase gene Proteins 0.000 description 3
- 206010057248 Cell death Diseases 0.000 description 3
- 108700010070 Codon Usage Proteins 0.000 description 3
- 239000003155 DNA primer Substances 0.000 description 3
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 3
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 3
- 241000283973 Oryctolagus cuniculus Species 0.000 description 3
- 108010058846 Ovalbumin Proteins 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 238000005571 anion exchange chromatography Methods 0.000 description 3
- AIYUHDOJVYHVIT-UHFFFAOYSA-M caesium chloride Chemical compound [Cl-].[Cs+] AIYUHDOJVYHVIT-UHFFFAOYSA-M 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 230000009089 cytolysis Effects 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 238000012217 deletion Methods 0.000 description 3
- 230000037430 deletion Effects 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 239000002751 oligonucleotide probe Substances 0.000 description 3
- 229940092253 ovalbumin Drugs 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 238000013492 plasmid preparation Methods 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 229940002612 prodrug Drugs 0.000 description 3
- 239000000651 prodrug Substances 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 230000028043 self proteolysis Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- 238000004809 thin layer chromatography Methods 0.000 description 3
- 238000013519 translation Methods 0.000 description 3
- 210000005253 yeast cell Anatomy 0.000 description 3
- GUPXYSSGJWIURR-UHFFFAOYSA-N 3-octoxypropane-1,2-diol Chemical compound CCCCCCCCOCC(O)CO GUPXYSSGJWIURR-UHFFFAOYSA-N 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 101000977023 Azospirillum brasilense Uncharacterized 17.8 kDa protein in nodG 5'region Proteins 0.000 description 2
- 101000961984 Bacillus thuringiensis Uncharacterized 30.3 kDa protein Proteins 0.000 description 2
- 108010038061 Chymotrypsinogen Proteins 0.000 description 2
- 238000001712 DNA sequencing Methods 0.000 description 2
- 101000644901 Drosophila melanogaster Putative 115 kDa protein in type-1 retrotransposable element R1DM Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 101000747702 Enterobacteria phage N4 Uncharacterized protein Gp2 Proteins 0.000 description 2
- 101000758599 Escherichia coli Uncharacterized 14.7 kDa protein Proteins 0.000 description 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- 101000768930 Lactococcus lactis subsp. cremoris Uncharacterized protein in pepC 5'region Proteins 0.000 description 2
- 101000976302 Leptospira interrogans Uncharacterized protein in sph 3'region Proteins 0.000 description 2
- 101000778886 Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai (strain 56601) Uncharacterized protein LA_2151 Proteins 0.000 description 2
- JOCBASBOOFNAJA-UHFFFAOYSA-N N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid Chemical compound OCC(CO)(CO)NCCS(O)(=O)=O JOCBASBOOFNAJA-UHFFFAOYSA-N 0.000 description 2
- 239000000020 Nitrocellulose Substances 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 2
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 108010033276 Peptide Fragments Proteins 0.000 description 2
- 102000007079 Peptide Fragments Human genes 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 239000012614 Q-Sepharose Substances 0.000 description 2
- 108020004511 Recombinant DNA Proteins 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 101001121571 Rice tungro bacilliform virus (isolate Philippines) Protein P2 Proteins 0.000 description 2
- 101000818098 Spirochaeta aurantia Uncharacterized protein in trpE 3'region Proteins 0.000 description 2
- 241000187747 Streptomyces Species 0.000 description 2
- 101001026590 Streptomyces cinnamonensis Putative polyketide beta-ketoacyl synthase 2 Proteins 0.000 description 2
- 102000019197 Superoxide Dismutase Human genes 0.000 description 2
- 108010012715 Superoxide dismutase Proteins 0.000 description 2
- 101000750896 Synechococcus elongatus (strain PCC 7942 / FACHB-805) Uncharacterized protein Synpcc7942_2318 Proteins 0.000 description 2
- 239000007994 TES buffer Substances 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
- IXKSXJFAGXLQOQ-XISFHERQSA-N WHWLQLKPGQPMY Chemical compound C([C@@H](C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)C1=CNC=N1 IXKSXJFAGXLQOQ-XISFHERQSA-N 0.000 description 2
- 101000916321 Xenopus laevis Transposon TX1 uncharacterized 149 kDa protein Proteins 0.000 description 2
- 101000760088 Zymomonas mobilis subsp. mobilis (strain ATCC 10988 / DSM 424 / LMG 404 / NCIMB 8938 / NRRL B-806 / ZM1) 20.9 kDa protein Proteins 0.000 description 2
- FJWGYAHXMCUOOM-QHOUIDNNSA-N [(2s,3r,4s,5r,6r)-2-[(2r,3r,4s,5r,6s)-4,5-dinitrooxy-2-(nitrooxymethyl)-6-[(2r,3r,4s,5r,6s)-4,5,6-trinitrooxy-2-(nitrooxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-3,5-dinitrooxy-6-(nitrooxymethyl)oxan-4-yl] nitrate Chemical compound O([C@@H]1O[C@@H]([C@H]([C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O)O[C@H]1[C@@H]([C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@@H](CO[N+]([O-])=O)O1)O[N+]([O-])=O)CO[N+](=O)[O-])[C@@H]1[C@@H](CO[N+]([O-])=O)O[C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O FJWGYAHXMCUOOM-QHOUIDNNSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000003277 amino acid sequence analysis Methods 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 2
- 238000011319 anticancer therapy Methods 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 229940041181 antineoplastic drug Drugs 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 229940104302 cytosine Drugs 0.000 description 2
- 229940127089 cytotoxic agent Drugs 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000003623 enhancer Substances 0.000 description 2
- 238000012869 ethanol precipitation Methods 0.000 description 2
- 229960005542 ethidium bromide Drugs 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 238000002523 gelfiltration Methods 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 229920001220 nitrocellulos Polymers 0.000 description 2
- 229940079938 nitrocellulose Drugs 0.000 description 2
- 238000002515 oligonucleotide synthesis Methods 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 229920002401 polyacrylamide Polymers 0.000 description 2
- 229910000160 potassium phosphate Inorganic materials 0.000 description 2
- 235000011009 potassium phosphates Nutrition 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000003156 radioimmunoprecipitation Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 229940035893 uracil Drugs 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- XOERABXRVAVTCE-UHFFFAOYSA-N 1,3-dimethyl-1-phenylthiourea Chemical compound CNC(=S)N(C)C1=CC=CC=C1 XOERABXRVAVTCE-UHFFFAOYSA-N 0.000 description 1
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 1
- 108010039627 Aprotinin Proteins 0.000 description 1
- 241000972773 Aulopiformes Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000701822 Bovine papillomavirus Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 108091033380 Coding strand Proteins 0.000 description 1
- 229920002271 DEAE-Sepharose Polymers 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 102100037101 Deoxycytidylate deaminase Human genes 0.000 description 1
- 108010016626 Dipeptides Proteins 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 108091029865 Exogenous DNA Proteins 0.000 description 1
- 108010058643 Fungal Proteins Proteins 0.000 description 1
- 101150038242 GAL10 gene Proteins 0.000 description 1
- 102100024637 Galectin-10 Human genes 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 1
- 241000235649 Kluyveromyces Species 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 101000929494 Mus musculus Adenosine deaminase Proteins 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 102000004140 Oncostatin M Human genes 0.000 description 1
- 108090000630 Oncostatin M Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 241000589538 Pseudomonas fragi Species 0.000 description 1
- 230000004570 RNA-binding Effects 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 1
- 239000012506 Sephacryl® Substances 0.000 description 1
- 241000702208 Shigella phage SfX Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 229960004405 aprotinin Drugs 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 108010058966 bacteriophage T7 induced DNA polymerase Proteins 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 238000012742 biochemical analysis Methods 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000012496 blank sample Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 229910052792 caesium Inorganic materials 0.000 description 1
- TVFDJXOCXUVLDH-UHFFFAOYSA-N caesium atom Chemical compound [Cs] TVFDJXOCXUVLDH-UHFFFAOYSA-N 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 239000007795 chemical reaction product 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
- 229960003677 chloroquine Drugs 0.000 description 1
- 238000011210 chromatographic step Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000012761 co-transfection Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 231100000599 cytotoxic agent Toxicity 0.000 description 1
- 108010015012 dCMP deaminase Proteins 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- KAKKHKRHCKCAGH-UHFFFAOYSA-L disodium;(4-nitrophenyl) phosphate;hexahydrate Chemical compound O.O.O.O.O.O.[Na+].[Na+].[O-][N+](=O)C1=CC=C(OP([O-])([O-])=O)C=C1 KAKKHKRHCKCAGH-UHFFFAOYSA-L 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000009585 enzyme analysis Methods 0.000 description 1
- 238000001952 enzyme assay Methods 0.000 description 1
- 239000006167 equilibration buffer Substances 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
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 235000013861 fat-free Nutrition 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000008103 glucose Substances 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
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 229940127121 immunoconjugate Drugs 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 238000001738 isopycnic centrifugation Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 229960004452 methionine Drugs 0.000 description 1
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 1
- FYFFGSSZFBZTAH-UHFFFAOYSA-N methylaminomethanetriol Chemical compound CNC(O)(O)O FYFFGSSZFBZTAH-UHFFFAOYSA-N 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 description 1
- 230000012666 negative regulation of transcription by glucose Effects 0.000 description 1
- 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 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000007899 nucleic acid hybridization Methods 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 238000002205 phenol-chloroform extraction Methods 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 230000001323 posttranslational effect Effects 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 230000009465 prokaryotic expression Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 230000006337 proteolytic cleavage Effects 0.000 description 1
- 230000004147 pyrimidine metabolism Effects 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000012146 running buffer Substances 0.000 description 1
- 235000019515 salmon Nutrition 0.000 description 1
- 229920006298 saran Polymers 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 239000004017 serum-free culture medium Substances 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 239000007974 sodium acetate buffer Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 1
- 230000005026 transcription initiation Effects 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 238000003151 transfection method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Enzymes And Modification Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US36502089A | 1989-06-09 | 1989-06-09 | |
| US07/531,646 US5338678A (en) | 1989-06-09 | 1990-06-01 | Expression of DNA sequences encoding a thermally stable cytosine deaminase from saccharomyces |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| NO902551D0 NO902551D0 (no) | 1990-06-08 |
| NO902551L NO902551L (no) | 1990-12-10 |
| NO301487B1 true NO301487B1 (no) | 1997-11-03 |
Family
ID=27002759
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| NO902551A NO301487B1 (no) | 1989-06-09 | 1990-06-08 | Isolert nukleinsyremolekyl kodende for en termisk stabil cytosin deaminase, rekombinant ekspresjonsvektor, vertscelle og fremgangsmåte for fremstilling av rekombinant termisk stabil cytosin deaminase |
Country Status (21)
| Country | Link |
|---|---|
| US (1) | US5545548A (pt) |
| EP (1) | EP0402108B1 (pt) |
| JP (1) | JP3001933B2 (pt) |
| CN (1) | CN1052263C (pt) |
| AT (1) | ATE118040T1 (pt) |
| CA (1) | CA2018273C (pt) |
| CZ (1) | CZ283492B6 (pt) |
| DE (1) | DE69016502T2 (pt) |
| DK (1) | DK0402108T3 (pt) |
| ES (1) | ES2070277T3 (pt) |
| FI (1) | FI95046C (pt) |
| GR (1) | GR3015207T3 (pt) |
| IE (1) | IE66054B1 (pt) |
| IL (1) | IL94649A (pt) |
| MY (1) | MY105816A (pt) |
| NO (1) | NO301487B1 (pt) |
| NZ (1) | NZ233949A (pt) |
| OA (1) | OA09214A (pt) |
| PL (1) | PL165149B1 (pt) |
| PT (1) | PT94331B (pt) |
| YU (1) | YU112090A (pt) |
Families Citing this family (23)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6555370B1 (en) | 1990-11-13 | 2003-04-29 | Immunex Corporation | Bifunctional selectable fusion genes |
| US5358866A (en) * | 1991-07-03 | 1994-10-25 | The United States Of America As Represented By The Department Of Health And Human Services | Cytosine deaminase negative selection system for gene transfer techniques and therapies |
| JPH09500783A (ja) * | 1993-05-21 | 1997-01-28 | ターゲッティッド ジェネティクス コーポレイション | シトシンデアミナーゼ（ｃｄ）遺伝子に基づく二機能性選択融合遺伝子 |
| US6451571B1 (en) | 1994-05-02 | 2002-09-17 | University Of Washington | Thymidine kinase mutants |
| US5856153A (en) * | 1994-11-17 | 1999-01-05 | Cayla | Suicide genes and new associations of pyrimidine nucleobase and nucleoside analogs with new suicide genes for gene therapy of acquired diseases |
| CA2243985C (en) | 1997-09-11 | 2004-08-10 | F. Hoffmann-La Roche Ag | Thermostable dna polymerases incorporating nucleoside triphosphates labeled with fluorescein family dyes |
| GB9810752D0 (en) * | 1998-05-19 | 1998-07-15 | Glaxo Group Ltd | Cystosine deaminase gene |
| US6962702B2 (en) | 1998-06-22 | 2005-11-08 | Immunomedics Inc. | Production and use of novel peptide-based agents for use with bi-specific antibodies |
| US8716558B2 (en) | 1999-06-30 | 2014-05-06 | Marker Gene Technologies, Inc. | Method of altering glycosylation of proteins in response to nojirimycin glucuronide in a plant cell expressing glucuronidase |
| US6656917B1 (en) * | 1999-06-30 | 2003-12-02 | Marker Gene Technologies, Inc. | Compositions and methods for targeted enzymatic release of cell regulatory compounds |
| US20060147420A1 (en) | 2004-03-10 | 2006-07-06 | Juan Fueyo | Oncolytic adenovirus armed with therapeutic genes |
| KR101690826B1 (ko) | 2002-07-15 | 2016-12-28 | 보드 오브 리전츠 더 유니버시티 오브 텍사스 시스템 | 포스파티딜에탄올아민에 결합하는 펩타이드, 및 바이러스 감염을 치료하는데 있어서 이의 용도 |
| US7534585B2 (en) | 2003-07-21 | 2009-05-19 | Transgene S.A. | Multifunctional cytokines |
| CA2532783C (en) * | 2003-07-21 | 2013-09-17 | Transgene S.A. | Polypeptide having an improved cytosine deaminase activity |
| US7109304B2 (en) | 2003-07-31 | 2006-09-19 | Immunomedics, Inc. | Humanized anti-CD19 antibodies |
| WO2005105991A1 (ja) * | 2004-04-28 | 2005-11-10 | Amano Enzyme Inc. | 放線菌由来のａｍｐデアミナーゼ、及びその使用 |
| EA034136B1 (ru) | 2007-11-09 | 2020-01-09 | Перигрин Фармасьютикалз, Инк. | Изолированное антитело, которое связывается с vegf, и его применения |
| US20090263884A1 (en) * | 2008-04-22 | 2009-10-22 | Organica Biotech, Inc. | Multi-action drain cleaning composition and method |
| CN105624193B (zh) | 2008-09-26 | 2021-07-13 | 索元生物医药(美国)有限公司 | 重组载体 |
| CN103031296B (zh) * | 2012-12-10 | 2014-12-17 | 浙江工业大学 | 冬虫夏草dCMP脱氨酶、编码基因及其应用 |
| CN103045573B (zh) * | 2012-12-10 | 2015-03-18 | 浙江工业大学 | 冬虫夏草胞嘧啶脱氨酶、编码基因及其应用 |
| CN109207535B (zh) * | 2018-09-20 | 2021-06-25 | 新乡拓新药业股份有限公司 | 利用铜绿假单胞菌合成尿嘧啶的方法 |
| CN116143947B (zh) * | 2023-01-16 | 2024-05-24 | 中国科学院青岛生物能源与过程研究所 | 一种酵母胞嘧啶脱氨酶融合蛋白及其突变体和应用 |
-
1990
- 1990-06-05 CA CA002018273A patent/CA2018273C/en not_active Expired - Lifetime
- 1990-06-06 IL IL9464990A patent/IL94649A/en active IP Right Grant
- 1990-06-06 AT AT90306131T patent/ATE118040T1/de not_active IP Right Cessation
- 1990-06-06 DE DE69016502T patent/DE69016502T2/de not_active Expired - Lifetime
- 1990-06-06 DK DK90306131.5T patent/DK0402108T3/da active
- 1990-06-06 EP EP90306131A patent/EP0402108B1/en not_active Expired - Lifetime
- 1990-06-06 NZ NZ233949A patent/NZ233949A/en unknown
- 1990-06-06 ES ES90306131T patent/ES2070277T3/es not_active Expired - Lifetime
- 1990-06-07 FI FI902855A patent/FI95046C/fi active IP Right Grant
- 1990-06-08 YU YU01120/90A patent/YU112090A/xx unknown
- 1990-06-08 CZ CS902865A patent/CZ283492B6/cs not_active IP Right Cessation
- 1990-06-08 OA OA59803A patent/OA09214A/xx unknown
- 1990-06-08 IE IE207990A patent/IE66054B1/en not_active IP Right Cessation
- 1990-06-08 PT PT94331A patent/PT94331B/pt not_active IP Right Cessation
- 1990-06-08 NO NO902551A patent/NO301487B1/no not_active IP Right Cessation
- 1990-06-08 MY MYPI90000950A patent/MY105816A/en unknown
- 1990-06-08 PL PL90285552A patent/PL165149B1/pl unknown
- 1990-06-09 CN CN90104922A patent/CN1052263C/zh not_active Expired - Lifetime
- 1990-06-11 JP JP2153799A patent/JP3001933B2/ja not_active Expired - Lifetime
-
1994
- 1994-06-20 US US08/262,238 patent/US5545548A/en not_active Expired - Lifetime
-
1995
- 1995-02-28 GR GR940403976T patent/GR3015207T3/el unknown
Also Published As
| Publication number | Publication date |
|---|---|
| CA2018273A1 (en) | 1990-12-09 |
| JPH04131084A (ja) | 1992-05-01 |
| US5545548A (en) | 1996-08-13 |
| DK0402108T3 (da) | 1995-04-10 |
| CN1052263C (zh) | 2000-05-10 |
| CA2018273C (en) | 1999-04-06 |
| PL285552A1 (en) | 1991-02-25 |
| ES2070277T3 (es) | 1995-06-01 |
| PT94331A (pt) | 1991-03-20 |
| ATE118040T1 (de) | 1995-02-15 |
| OA09214A (en) | 1992-06-30 |
| JP3001933B2 (ja) | 2000-01-24 |
| FI95046B (fi) | 1995-08-31 |
| FI902855A0 (fi) | 1990-06-07 |
| YU112090A (en) | 1992-05-28 |
| PT94331B (pt) | 1997-04-30 |
| NO902551D0 (no) | 1990-06-08 |
| IE66054B1 (en) | 1995-12-13 |
| PL165149B1 (pl) | 1994-11-30 |
| EP0402108A1 (en) | 1990-12-12 |
| EP0402108B1 (en) | 1995-02-01 |
| CS286590A3 (en) | 1992-02-19 |
| DE69016502T2 (de) | 1995-06-01 |
| NZ233949A (en) | 1992-06-25 |
| CN1050221A (zh) | 1991-03-27 |
| GR3015207T3 (en) | 1995-05-31 |
| FI95046C (fi) | 1995-12-11 |
| IE902079L (en) | 1990-12-09 |
| IL94649A (en) | 1995-11-27 |
| DE69016502D1 (de) | 1995-03-16 |
| NO902551L (no) | 1990-12-10 |
| CZ283492B6 (cs) | 1998-04-15 |
| MY105816A (en) | 1995-01-30 |
| IL94649A0 (en) | 1991-04-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5338678A (en) | Expression of DNA sequences encoding a thermally stable cytosine deaminase from saccharomyces | |
| NO301487B1 (no) | Isolert nukleinsyremolekyl kodende for en termisk stabil cytosin deaminase, rekombinant ekspresjonsvektor, vertscelle og fremgangsmåte for fremstilling av rekombinant termisk stabil cytosin deaminase | |
| Cain et al. | Interaction between Glu-219 and His-245 within the a subunit of F1F0-ATPase in Escherichia coli. | |
| JP4450767B2 (ja) | サイクリックｇｍｐ結合性、サイクリックｇｍｐ特異的ホスホジエステラーゼの材料および方法 | |
| CN110373404B (zh) | 一种酶突变体 | |
| Van der Linden et al. | Possible role of Escherichia coli penicillin-binding protein 6 in stabilization of stationary-phase peptidoglycan | |
| GIRAUD et al. | ATP synthase of yeast mitochondria: Isolation of the F1δ subunit, sequence and disruption of the structural gene | |
| US6406899B1 (en) | Highly active alkaline phosphatase | |
| JPH02177887A (ja) | グルコース―６―リン酸デヒドロゲナーゼ遺伝子 | |
| US5728806A (en) | TIH1, protein that interacts with casein kinase I | |
| Hayden et al. | Cloning, Overexpression, and Purification of Cytosine Deaminase fromSaccharomyces cerevisiae | |
| EP0445097B1 (en) | Method for protein N-myristoylation | |
| Benaroudj et al. | Overexpression in Escherichia coli, purification and characterization of the molecular chaperone HSC70 | |
| JP3486942B2 (ja) | 耐熱性エステラーゼ | |
| Pan et al. | Sequence analysis and expression of phospholipase A2 from Taiwan cobra | |
| JPH05192163A (ja) | イノシトールデヒドロゲナーゼ遺伝子 | |
| CA2232841A1 (en) | Process for producing natriuretic peptides via streptavidine fusion proteins | |
| Keßler et al. | Ser644 is important for catalytic activity but is not involved in cAMP-dependent phosphorylation of yeast 6-phosphofructo-2-kinase | |
| Aoki et al. | Modification of human neuron‐specific enolase for application to radioimmunoassay | |
| JPH02308793A (ja) | ミコバクテリウム・カンサシ由来のα抗原 | |
| JPH05276960A (ja) | アスパラギニルエンドペプチダーゼ遺伝子 | |
| JPH07274969A (ja) | アーモンドｎ−グリコシダーゼ遺伝子 | |
| JPH11318464A (ja) | 耐熱性ｐｐｉ及びそれをコードする遺伝子 | |
| JPH06253857A (ja) | 遺伝子およびその形質転換体ならびにポリペプチドの製造方法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MK1K | Patent expired |