FI107265B - Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster - Google Patents
Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster Download PDFInfo
- Publication number
- FI107265B FI107265B FI992546A FI19992546A FI107265B FI 107265 B FI107265 B FI 107265B FI 992546 A FI992546 A FI 992546A FI 19992546 A FI19992546 A FI 19992546A FI 107265 B FI107265 B FI 107265B
- Authority
- FI
- Finland
- Prior art keywords
- gene
- isolated
- aox1
- dna
- pichia pastoris
- Prior art date
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 136
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims description 39
- 102000004169 proteins and genes Human genes 0.000 title abstract description 29
- 208000002672 hepatitis B Diseases 0.000 title abstract description 12
- 239000013598 vector Substances 0.000 claims description 81
- 239000012634 fragment Substances 0.000 claims description 69
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 38
- 230000001105 regulatory effect Effects 0.000 claims description 26
- 241000235058 Komagataella pastoris Species 0.000 claims description 23
- 239000003550 marker Substances 0.000 claims description 19
- 101150069554 HIS4 gene Proteins 0.000 claims description 16
- 241000235648 Pichia Species 0.000 claims description 14
- 101150005709 ARG4 gene Proteins 0.000 claims description 7
- 101150061183 AOX1 gene Proteins 0.000 claims description 6
- 101150010882 S gene Proteins 0.000 claims description 3
- 101100004044 Vigna radiata var. radiata AUX22B gene Proteins 0.000 claims description 3
- 102100036826 Aldehyde oxidase Human genes 0.000 claims 9
- 101000928314 Homo sapiens Aldehyde oxidase Proteins 0.000 claims 9
- 101710093617 Dihydroxyacetone synthase Proteins 0.000 claims 2
- 101150030170 Paat gene Proteins 0.000 claims 1
- 229940037201 oris Drugs 0.000 claims 1
- UNXRWKVEANCORM-UHFFFAOYSA-I triphosphate(5-) Chemical compound [O-]P([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O UNXRWKVEANCORM-UHFFFAOYSA-I 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 51
- 239000002245 particle Substances 0.000 abstract description 25
- 102100031673 Corneodesmosin Human genes 0.000 abstract description 16
- 101710139375 Corneodesmosin Proteins 0.000 abstract description 15
- 230000000890 antigenic effect Effects 0.000 abstract description 11
- 238000004519 manufacturing process Methods 0.000 abstract description 11
- 239000013612 plasmid Substances 0.000 description 91
- 108020004414 DNA Proteins 0.000 description 71
- 210000004027 cell Anatomy 0.000 description 40
- 238000006460 hydrolysis reaction Methods 0.000 description 34
- 230000007062 hydrolysis Effects 0.000 description 33
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 31
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 30
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 30
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 28
- 235000018102 proteins Nutrition 0.000 description 27
- 241000700721 Hepatitis B virus Species 0.000 description 26
- 230000009466 transformation Effects 0.000 description 25
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 24
- 239000000523 sample Substances 0.000 description 22
- 239000000243 solution Substances 0.000 description 18
- 108010025188 Alcohol oxidase Proteins 0.000 description 16
- 235000001014 amino acid Nutrition 0.000 description 16
- 150000001413 amino acids Chemical class 0.000 description 16
- 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 16
- 229960000723 ampicillin Drugs 0.000 description 16
- 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 15
- 238000006243 chemical reaction Methods 0.000 description 15
- 239000011780 sodium chloride Substances 0.000 description 15
- 108010054576 Deoxyribonuclease EcoRI Proteins 0.000 description 14
- 241000588724 Escherichia coli Species 0.000 description 14
- 108091007433 antigens Proteins 0.000 description 14
- 239000006228 supernatant Substances 0.000 description 14
- 108091028043 Nucleic acid sequence Proteins 0.000 description 13
- 108091034117 Oligonucleotide Proteins 0.000 description 13
- 239000000427 antigen Substances 0.000 description 13
- 102000036639 antigens Human genes 0.000 description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- 241001288713 Escherichia coli MC1061 Species 0.000 description 12
- 239000008188 pellet Substances 0.000 description 12
- 238000009396 hybridization Methods 0.000 description 11
- 101100173636 Rattus norvegicus Fhl2 gene Proteins 0.000 description 10
- 239000000872 buffer Substances 0.000 description 10
- 239000000203 mixture Substances 0.000 description 10
- 238000002360 preparation method Methods 0.000 description 10
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 230000001580 bacterial effect Effects 0.000 description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 8
- 108010042407 Endonucleases Proteins 0.000 description 8
- 102000004533 Endonucleases Human genes 0.000 description 8
- 102000003960 Ligases Human genes 0.000 description 8
- 108090000364 Ligases Proteins 0.000 description 8
- 210000004899 c-terminal region Anatomy 0.000 description 8
- 238000010276 construction Methods 0.000 description 8
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 8
- 238000001502 gel electrophoresis Methods 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- 238000002703 mutagenesis Methods 0.000 description 8
- 231100000350 mutagenesis Toxicity 0.000 description 8
- 239000011543 agarose gel Substances 0.000 description 7
- 238000000605 extraction Methods 0.000 description 7
- 230000010076 replication Effects 0.000 description 7
- 108020004705 Codon Proteins 0.000 description 6
- 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 6
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 6
- 230000000295 complement effect Effects 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 6
- 230000004151 fermentation Effects 0.000 description 6
- 208000015181 infectious disease Diseases 0.000 description 6
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 241000235070 Saccharomyces Species 0.000 description 5
- 239000007983 Tris buffer Substances 0.000 description 5
- 239000011324 bead Substances 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 238000003780 insertion Methods 0.000 description 5
- 230000037431 insertion Effects 0.000 description 5
- 230000010354 integration Effects 0.000 description 5
- 238000002955 isolation Methods 0.000 description 5
- 239000002773 nucleotide Substances 0.000 description 5
- 125000003729 nucleotide group Chemical group 0.000 description 5
- 229940063673 spermidine Drugs 0.000 description 5
- 230000001131 transforming effect Effects 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- 229920000936 Agarose Polymers 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 238000000246 agarose gel electrophoresis Methods 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 238000001962 electrophoresis Methods 0.000 description 4
- 238000012869 ethanol precipitation Methods 0.000 description 4
- 230000003301 hydrolyzing effect Effects 0.000 description 4
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 229920001223 polyethylene glycol Polymers 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- 229960005486 vaccine Drugs 0.000 description 4
- 108091026890 Coding region Proteins 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 3
- 102000012410 DNA Ligases Human genes 0.000 description 3
- 108010061982 DNA Ligases Proteins 0.000 description 3
- 108090001030 Lipoproteins Proteins 0.000 description 3
- 102000004895 Lipoproteins Human genes 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 3
- 241000364057 Peoria Species 0.000 description 3
- 108091000080 Phosphotransferase Proteins 0.000 description 3
- 108020004511 Recombinant DNA Proteins 0.000 description 3
- 239000007984 Tris EDTA buffer Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000002458 infectious effect Effects 0.000 description 3
- 239000006166 lysate Substances 0.000 description 3
- 229910001629 magnesium chloride Inorganic materials 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 239000010452 phosphate Substances 0.000 description 3
- 102000020233 phosphotransferase Human genes 0.000 description 3
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 239000011541 reaction mixture Substances 0.000 description 3
- 101150098466 rpsL gene Proteins 0.000 description 3
- 229940081969 saccharomyces cerevisiae Drugs 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- KMEMIMRPZGDOMG-UHFFFAOYSA-N 2-cyanoethoxyphosphonamidous acid Chemical compound NP(O)OCCC#N KMEMIMRPZGDOMG-UHFFFAOYSA-N 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 2
- 230000006820 DNA synthesis Effects 0.000 description 2
- RXKJFZQQPQGTFL-UHFFFAOYSA-N Dihydroxyacetone Natural products OCC(=O)CO RXKJFZQQPQGTFL-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 102100024319 Intestinal-type alkaline phosphatase Human genes 0.000 description 2
- 101710184243 Intestinal-type alkaline phosphatase Proteins 0.000 description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 2
- 239000006137 Luria-Bertani broth Substances 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 239000000020 Nitrocellulose Substances 0.000 description 2
- 108010021757 Polynucleotide 5'-Hydroxyl-Kinase Proteins 0.000 description 2
- 102000008422 Polynucleotide 5'-hydroxyl-kinase Human genes 0.000 description 2
- 229940096437 Protein S Drugs 0.000 description 2
- 101150014136 SUC2 gene Proteins 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 238000002105 Southern blotting Methods 0.000 description 2
- 208000037065 Subacute sclerosing leukoencephalitis Diseases 0.000 description 2
- 206010042297 Subacute sclerosing panencephalitis Diseases 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- -1 alpha pairing factor Chemical compound 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 235000011148 calcium chloride Nutrition 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 230000009089 cytolysis Effects 0.000 description 2
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 2
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 229940120503 dihydroxyacetone Drugs 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000002523 gelfiltration Methods 0.000 description 2
- 101150101299 gene 4 gene Proteins 0.000 description 2
- 238000010362 genome editing Methods 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 208000006454 hepatitis Diseases 0.000 description 2
- 231100000283 hepatitis Toxicity 0.000 description 2
- 101150050908 hisA gene Proteins 0.000 description 2
- 235000014304 histidine Nutrition 0.000 description 2
- 101150085823 hsdR gene Proteins 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 230000002934 lysing effect Effects 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 229930182817 methionine Natural products 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 229920001220 nitrocellulos Polymers 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 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
- OJMIONKXNSYLSR-UHFFFAOYSA-N phosphorous acid Chemical compound OP(O)O OJMIONKXNSYLSR-UHFFFAOYSA-N 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- SCVFZCLFOSHCOH-UHFFFAOYSA-M potassium acetate Chemical compound [K+].CC([O-])=O SCVFZCLFOSHCOH-UHFFFAOYSA-M 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 101150077403 priA gene Proteins 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000017854 proteolysis Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 241001515965 unidentified phage Species 0.000 description 2
- 210000002845 virion Anatomy 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 210000005253 yeast cell Anatomy 0.000 description 2
- LXJXRIRHZLFYRP-VKHMYHEASA-L (R)-2-Hydroxy-3-(phosphonooxy)-propanal Natural products O=C[C@H](O)COP([O-])([O-])=O LXJXRIRHZLFYRP-VKHMYHEASA-L 0.000 description 1
- GZCWLCBFPRFLKL-UHFFFAOYSA-N 1-prop-2-ynoxypropan-2-ol Chemical compound CC(O)COCC#C GZCWLCBFPRFLKL-UHFFFAOYSA-N 0.000 description 1
- 125000001731 2-cyanoethyl group Chemical group [H]C([H])(*)C([H])([H])C#N 0.000 description 1
- OPIFSICVWOWJMJ-AEOCFKNESA-N 5-bromo-4-chloro-3-indolyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 OPIFSICVWOWJMJ-AEOCFKNESA-N 0.000 description 1
- 102000013563 Acid Phosphatase Human genes 0.000 description 1
- 108010051457 Acid Phosphatase Proteins 0.000 description 1
- 208000030090 Acute Disease Diseases 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 1
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 208000002109 Argyria Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108020000946 Bacterial DNA Proteins 0.000 description 1
- 240000002791 Brassica napus Species 0.000 description 1
- 235000004977 Brassica sinapistrum Nutrition 0.000 description 1
- 102100035893 CD151 antigen Human genes 0.000 description 1
- 101710118846 CD151 antigen Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 102100030497 Cytochrome c Human genes 0.000 description 1
- 108010075031 Cytochromes c Proteins 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- LXJXRIRHZLFYRP-VKHMYHEASA-N D-glyceraldehyde 3-phosphate Chemical compound O=C[C@H](O)COP(O)(O)=O LXJXRIRHZLFYRP-VKHMYHEASA-N 0.000 description 1
- 102000004594 DNA Polymerase I Human genes 0.000 description 1
- 108010017826 DNA Polymerase I Proteins 0.000 description 1
- 229920001917 Ficoll Polymers 0.000 description 1
- 102000048120 Galactokinases Human genes 0.000 description 1
- 108700023157 Galactokinases 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
- 239000004471 Glycine Substances 0.000 description 1
- 241001149669 Hanseniaspora Species 0.000 description 1
- 208000028523 Hereditary Complement Deficiency disease Diseases 0.000 description 1
- 101100351324 Homo sapiens PDPN gene Proteins 0.000 description 1
- 101000894590 Homo sapiens Uncharacterized protein C20orf85 Proteins 0.000 description 1
- 241001506991 Komagataella phaffii GS115 Species 0.000 description 1
- 206010067125 Liver injury Diseases 0.000 description 1
- 239000006142 Luria-Bertani Agar Substances 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 101100174763 Mus musculus Galk1 gene Proteins 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 241000320412 Ogataea angusta Species 0.000 description 1
- 240000007817 Olea europaea Species 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 1
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 1
- 235000011613 Pinus brutia Nutrition 0.000 description 1
- 241000018646 Pinus brutia Species 0.000 description 1
- 102100037265 Podoplanin Human genes 0.000 description 1
- 229920002562 Polyethylene Glycol 3350 Polymers 0.000 description 1
- 108010029485 Protein Isoforms Proteins 0.000 description 1
- 102000001708 Protein Isoforms Human genes 0.000 description 1
- 102000029301 Protein S Human genes 0.000 description 1
- 101000836384 Rattus norvegicus Serpin H1 Proteins 0.000 description 1
- 241000223252 Rhodotorula Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 101710137302 Surface antigen S Proteins 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 102100021442 Uncharacterized protein C20orf85 Human genes 0.000 description 1
- 241001105470 Valenzuela Species 0.000 description 1
- 235000009499 Vanilla fragrans Nutrition 0.000 description 1
- 244000263375 Vanilla tahitensis Species 0.000 description 1
- 235000012036 Vanilla tahitensis Nutrition 0.000 description 1
- 108010003533 Viral Envelope Proteins Proteins 0.000 description 1
- 108010031318 Vitronectin Proteins 0.000 description 1
- ZVQOOHYFBIDMTQ-UHFFFAOYSA-N [methyl(oxido){1-[6-(trifluoromethyl)pyridin-3-yl]ethyl}-lambda(6)-sulfanylidene]cyanamide Chemical compound N#CN=S(C)(=O)C(C)C1=CC=C(C(F)(F)F)N=C1 ZVQOOHYFBIDMTQ-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000005571 anion exchange chromatography Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 239000012131 assay buffer Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 108010051210 beta-Fructofuranosidase Proteins 0.000 description 1
- 108010005774 beta-Galactosidase Proteins 0.000 description 1
- 230000008238 biochemical pathway Effects 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 239000007975 buffered saline Substances 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 235000011089 carbon dioxide Nutrition 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 description 1
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 1
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 238000011026 diafiltration Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 101150012763 endA gene Proteins 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 101150041954 galU gene Proteins 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 101150096208 gtaB gene Proteins 0.000 description 1
- 231100000753 hepatic injury Toxicity 0.000 description 1
- SPSXSWRZQFPVTJ-ZQQKUFEYSA-N hepatitis b vaccine Chemical compound C([C@H](NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](CC1N=CN=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)OC(=O)CNC(=O)CNC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@@H](N)CCCNC(N)=N)C1=CC=CC=C1 SPSXSWRZQFPVTJ-ZQQKUFEYSA-N 0.000 description 1
- 229940124736 hepatitis-B vaccine Drugs 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 150000002411 histidines Chemical class 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000002198 insoluble material Substances 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
- 239000003446 ligand Substances 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000006151 minimal media Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 210000002445 nipple Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 229940050929 polyethylene glycol 3350 Drugs 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 235000011056 potassium acetate Nutrition 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 150000003138 primary alcohols Chemical class 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000009877 rendering Methods 0.000 description 1
- 238000002271 resection Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 239000012465 retentate Substances 0.000 description 1
- 101150072534 sbcB gene Proteins 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000006748 scratching Methods 0.000 description 1
- 230000002393 scratching effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011684 sodium molybdate Substances 0.000 description 1
- 235000015393 sodium molybdate Nutrition 0.000 description 1
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000002344 surface layer Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 229910052720 vanadium Inorganic materials 0.000 description 1
- 239000012808 vapor phase Substances 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 239000011686 zinc sulphate Substances 0.000 description 1
- 235000009529 zinc sulphate Nutrition 0.000 description 1
- 108010082737 zymolyase Proteins 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
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
- C12N15/815—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts for yeasts other than Saccharomyces
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
-
- 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
- C12N2730/00—Reverse transcribing DNA viruses
- C12N2730/00011—Details
- C12N2730/10011—Hepadnaviridae
- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
- C12N2730/10122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Claims (9)
1. Kombinationsvektor, kännetecknad av att den innefattar a) minst ett DNA-fragment som skall inserteras, och som är 15 ett funktionellt regleromräde av en 51-AOXl-gen. vars längd är ca en kilobas och som isolerats frän Pichia pastoris. funktionellt kopplat tili b) en strukturell gen av S, funktionellt kopplad tili c) 3'- terminal s elevens en av AOX1, som isolerats frän Pichia 20 pastoris. ligaterad tili • * • · « • · · ··· * .·. : d) en markörgen, som är ARG4, som isolerats frän Pichia pas- toris. ligaterad tili • · · • ·· *.· ; e) ett andra DNA-fragment som skall inserteras som är ca 0,65 kilobas av 3'-terminalsekvensen av AOX1: eller • * 25 a) minst ett DNA-fragment som skall inserteras, och som är ett funktionellt regleromräde av en 5'-AOXl-gen, vars längd är ca en kilobas och som isolerats frän Pichia pastoris. funktionellt kopplat tili -*·* b) en strukturell gen av preS2, funktionellt kopplad tili • · • · · • · · • · 107265 c) 31-terminalsekvensen av AOX1. som isolerats frän Pichia pastoria. ligaterad till d) en markorgen, som är HIS4, som isolerats frän Pichia pas-toris. ligaterad till 5 e) ett andra DNA-fragment som skall inserteras som är ca 0,65 kilobas av 31-terminalsekvensen av AOX1.
2. Metylotrofisk jäst, kännetecknad av att den transforme-rats med minst en expressionskassett, som innefattar ett reg-leromräde, som är funktionellt kopplat till en strukturell 10 gen av preS2, som är funktionellt kopplad till 3'-terminalsekvensen, och med minst en expressionskassett, som innefat-tar ett regleromräde, som är funktionellt kopplat till en strukturell gen av S, som är funktionellt kopplad till 3'-terminalsekvensen.
3. Metylotrofisk jäst enligt patentkrav 2, kännetecknad av att jästen är Pichia pastoris.
4. Metylotrofisk jäst enligt patentkrav 3, kännetecknad av att jästen är stam PPF1 av Pichia pastoris.
5. Metylotrofisk jäst enligt patentkrav 4, kännetecknad av 20 att nämnda Pichia pastoris PPF1 transformerats a) med minst en lineär, platsspecifik kombinationsvektor, • · :.: : som innef attar anordnade i serie I » · • «· « · i) ett första DNA-fragment som skall inserteras • · » • · · * ii) en markorgen och minst en Pichia-kompatibel expres 25 sionskassett, som innehäller en strukturell gen av preS2, som *:**: är funktionellt kopplad till iii) 31-terminalsekvensen, som väljs i gruppen bestäende av 3 1 - terminalsekvensen isolerad frän AOXl-genen, p40-gene:i, DHAS-genen och HIS4-genen, vilka isolerats frän Pichia pas :o- .1.1.30 ris. • · · « · • · · · 107265 iv) ett andra DNA-fragment som skall inserteras, varvid den inbördes ordningen mellan markorgenen och kasset-ten av komponent (ii) kan bytas; och b) med minst en platsspecifik kombinationsvektor, som inne-5 fattar foljande: i) minst ett DNA-fragment som skall inserteras ii) en markörgen och minst en Pichia-kompatibel expres-sionskassett, som innehäller en strukturell gen av S, som är funktionellt kopplad till 10 iii) 3 1-terminalsekvensen, som väljs i gruppen bestäende av 3'-terminalsekvensen isolerad frän AOXl-genen, p40-genen, DHAS-genen och HIS4-genen, vilka isolerats frän Pichia pasto-ris, varvid den inbördes ordningen mellan markörgenen och kasset-15 ten av komponent (ii) kan bytas.
6. Metylotrofisk jäst enligt patentkrav 5, kannetecknad av att kombinationsvektom under punkt (b) innefattar a) ett första DNA-fragment som skall inserteras, och som ligger ca en kilobas frän 5' -änden av AOXl-regleromrade, som |j* 20 isolerats frän Pichia pastoris. funktionellt kopplat till • · • « · • · « *1* \ b) en strukturell gen av preS^# funktionellt kopplad till • · · • · · • · : c) 31-terminalsekvensen av AOXl. som isolerats frän Pichia pastoris. ligaterad till . d) en markörgen, som är HIS4 isolerad frän Pichia pastoris. * ’ 25 ligaterad till e) ett andra DNA-fragment som skall inserteras som är ca 0,65 kilobas av 3'-änden av AOXl-terminalsekvensen.
. 7. Metylotrofisk jäst enligt patentkrav 5, kannetecknad av : att nämnda kombinationsvektor under punkt (a) innefattar • · 107265 a) ett första DNA-fragment som skall inserteras, och som ligger ca en kilobas frän 5'-änden av AOXl-regleromräde, som isolerats frän Pichia pastoria, funktionellt kopplat tili b) en strukturell gen av S, funktionellt kopplad tili 5 c) 3'-terminalsekvensen av AOX1. som isolerats frän Pichia pastoris. ligaterad tili d) en markörgen, som är ARG4 isolerad frän Saccharomyces ce-revisiae, ligaterad tili e) ett andra DNA-fragment som skall inserteras, som är ca 10 0,65 kilobas av 31-terminalsekvensen av AOX1.
8. Metylotrofisk jäst enligt patentkrav 5, kännetecknad av att nämnda Pichia pastoris PPF1 transformeras med flera än en kopia av nämnda lineära platsspecifika kombinationsvektor.
9. Pichia pastoris PPF1 (NRRL Y-18017) transformerad med :vä 15 kopior av en preS2-gen av pTB05A och en kopia av en S-gen (exempel II). « « « • · · · • · • · · • · · • ·· · • · • · » • · · • · • · · • · · • · · • · · • · · • · · • · rt · « * « I • «
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US19371488A | 1988-05-13 | 1988-05-13 | |
| US19371488 | 1988-05-13 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| FI19992546A FI19992546A (sv) | 1999-11-29 |
| FI107265B true FI107265B (sv) | 2001-06-29 |
Family
ID=22714736
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| FI892312A FI104639B (sv) | 1988-05-13 | 1989-05-12 | Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster |
| FI992546A FI107265B (sv) | 1988-05-13 | 1999-11-29 | Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| FI892312A FI104639B (sv) | 1988-05-13 | 1989-05-12 | Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster |
Country Status (25)
| Country | Link |
|---|---|
| US (2) | US5670630A (sv) |
| EP (1) | EP0341746B1 (sv) |
| JP (1) | JP2614314B2 (sv) |
| KR (1) | KR100193701B1 (sv) |
| CN (1) | CN1038668A (sv) |
| AT (1) | ATE151109T1 (sv) |
| AU (1) | AU618596B2 (sv) |
| CA (1) | CA1340617C (sv) |
| DD (1) | DD285373A5 (sv) |
| DE (1) | DE68927921T2 (sv) |
| DK (1) | DK235289A (sv) |
| ES (1) | ES2101676T3 (sv) |
| FI (2) | FI104639B (sv) |
| GR (1) | GR3023985T3 (sv) |
| HU (1) | HU207532B (sv) |
| IE (1) | IE81158B1 (sv) |
| IL (1) | IL90161A0 (sv) |
| IN (1) | IN171696B (sv) |
| MX (1) | MX26715A (sv) |
| NO (1) | NO300465B1 (sv) |
| NZ (1) | NZ228948A (sv) |
| PL (1) | PL161997B1 (sv) |
| PT (1) | PT90553A (sv) |
| YU (1) | YU98689A (sv) |
| ZA (1) | ZA893440B (sv) |
Families Citing this family (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5032516A (en) * | 1985-10-25 | 1991-07-16 | Phillips Petroleum Company | Pichia pastoris alcohol oxidase II regulatory region |
| PL168787B1 (pl) * | 1989-08-03 | 1996-04-30 | Smithkline Beecham Biolog | Sposób wytwarzania czastki kompozytowej PL |
| GB9007416D0 (en) * | 1990-04-02 | 1990-05-30 | Wellcome Found | Expression of heterologous protein in yeast |
| US6197548B1 (en) | 1990-04-02 | 2001-03-06 | Medeva Pharma Limited | Transformed Pichia expressing the pertactin antigen |
| EP0521348A1 (en) * | 1991-06-18 | 1993-01-07 | Korea Institute Of Science And Technology | Chimeric antibody with specificity for hepatitis B virus pre-S2 surface antigen and cell line producing same |
| LT3988B (en) | 1992-02-17 | 1996-06-25 | Fermentas Biotech Inst | Recombinant plasmides pfs19, pfps2-48 and pjlfds1 codingsynthesis of human hepatite b of surfice virus antigenes, methods fof producing thereof |
| CA2101610A1 (en) * | 1992-08-07 | 1994-02-08 | William D. Prevatt | Production of bacillus entomotoxins in methylotrophic yeast |
| GB9402832D0 (en) * | 1994-02-15 | 1994-04-06 | Smith Kline Beecham Biolog S A | Novel process |
| US5955349A (en) * | 1996-08-26 | 1999-09-21 | Zymogenetics, Inc. | Compositions and methods for producing heterologous polypeptides in Pichia methanolica |
| EP0889966A1 (en) * | 1995-11-09 | 1999-01-13 | ZymoGenetics, Inc. | Compositions and methods for producing heterologous polypeptides in pichia methanolica |
| US5716808A (en) * | 1995-11-09 | 1998-02-10 | Zymogenetics, Inc. | Genetic engineering of pichia methanolica |
| MX9605082A (es) | 1996-10-24 | 1998-04-30 | Univ Autonoma De Nuevo Leon | Levaduras metilotroficas modificadas geneticamente para la produccion y secrecion de hormona de crecimiento humano. |
| JP2008500364A (ja) * | 2004-05-25 | 2008-01-10 | キメラコア， インコーポレイテッド | 自己集合性ナノ粒子薬物送達システム |
| CN1704475A (zh) * | 2004-05-31 | 2005-12-07 | 成都生物制品研究所 | 羧端含前s2免疫决定簇的乙肝表面抗原融合基因及蛋白质 |
| EP1773861A4 (en) * | 2004-07-22 | 2008-04-16 | Glycofi Inc | MALATE SYNTHASE REGULATION SEQUENCES FOR THE EXPRESSION OF A HETEROLOGOUS GENE IN PICHIA |
| CN100381171C (zh) * | 2004-12-30 | 2008-04-16 | 成都生物制品研究所 | 含前s1、前s2和s抗原决定簇的乙肝表面抗原复合颗粒 |
| CA2648581A1 (en) * | 2006-04-07 | 2008-09-12 | Chimeros, Inc. | Compositions and methods for treating b- cell malignancies |
| WO2008124165A2 (en) * | 2007-04-09 | 2008-10-16 | Chimeros, Inc. | Self-assembling nanoparticle drug delivery system |
| WO2010120874A2 (en) | 2009-04-14 | 2010-10-21 | Chimeros, Inc. | Chimeric therapeutics, compositions, and methods for using same |
| AU2012305714A1 (en) | 2011-09-09 | 2014-03-27 | Biomed Realty, L.P. | Methods and compositions for controlling assembly of viral proteins |
| ES2926021T3 (es) | 2012-10-23 | 2022-10-21 | Toolgen Inc | Composición para escindir un ADN objetivo que comprende un ARN guía específico para el ADN objetivo y ácido nucleico codificador de proteína Cas o proteína Cas, y uso de la misma |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GR76274B (sv) * | 1981-08-04 | 1984-08-04 | Univ California | |
| EP0401941A3 (en) * | 1984-07-11 | 1991-04-17 | Takeda Chemical Industries, Ltd. | Hepatitis b virus surface antigen and production thereof |
| US4895800A (en) * | 1985-11-26 | 1990-01-23 | Phillips Petroleum Company | Yeast production of hepatitis B surface antigen |
| US4816564A (en) * | 1986-01-31 | 1989-03-28 | Merck & Co., Inc. | Method for producing hepatitis B virus proteins in yeast |
| JPS62236496A (ja) * | 1986-04-07 | 1987-10-16 | Green Cross Corp:The | ＨＢｓＡｇの製造方法 |
| US4963483A (en) * | 1987-10-13 | 1990-10-16 | Merck & Co., Inc. | Method for producing hepatitis B virus proteins in yeast |
| IL89991A0 (en) * | 1988-04-25 | 1989-12-15 | Phillips Petroleum Co | Expression of hepatitis b pres2 protein in methylotrophic yeasts |
-
1989
- 1989-05-02 NZ NZ228948A patent/NZ228948A/en unknown
- 1989-05-02 IL IL90161A patent/IL90161A0/xx unknown
- 1989-05-09 ZA ZA893440A patent/ZA893440B/xx unknown
- 1989-05-11 HU HU892372A patent/HU207532B/hu not_active IP Right Cessation
- 1989-05-11 KR KR1019890006430A patent/KR100193701B1/ko not_active IP Right Cessation
- 1989-05-11 NO NO891921A patent/NO300465B1/no not_active IP Right Cessation
- 1989-05-12 IE IE156389A patent/IE81158B1/en not_active IP Right Cessation
- 1989-05-12 EP EP89108632A patent/EP0341746B1/en not_active Expired - Lifetime
- 1989-05-12 DE DE68927921T patent/DE68927921T2/de not_active Expired - Lifetime
- 1989-05-12 AT AT89108632T patent/ATE151109T1/de not_active IP Right Cessation
- 1989-05-12 PT PT90553A patent/PT90553A/pt not_active Application Discontinuation
- 1989-05-12 AU AU34734/89A patent/AU618596B2/en not_active Expired
- 1989-05-12 FI FI892312A patent/FI104639B/sv active IP Right Grant
- 1989-05-12 PL PL89279426A patent/PL161997B1/pl unknown
- 1989-05-12 ES ES89108632T patent/ES2101676T3/es not_active Expired - Lifetime
- 1989-05-12 DK DK235289A patent/DK235289A/da not_active Application Discontinuation
- 1989-05-12 DD DD89328581A patent/DD285373A5/de unknown
- 1989-05-12 YU YU98689A patent/YU98689A/sh unknown
- 1989-05-12 MX MX2671589A patent/MX26715A/es unknown
- 1989-05-13 CN CN89103330A patent/CN1038668A/zh active Pending
- 1989-05-15 CA CA000599724A patent/CA1340617C/en not_active Expired - Lifetime
- 1989-05-15 JP JP1121313A patent/JP2614314B2/ja not_active Expired - Fee Related
- 1989-05-22 IN IN388/CAL/89A patent/IN171696B/en unknown
-
1995
- 1995-06-06 US US08/467,345 patent/US5670630A/en not_active Expired - Lifetime
- 1995-06-06 US US08/467,660 patent/US5650296A/en not_active Expired - Lifetime
-
1997
- 1997-07-02 GR GR970401639T patent/GR3023985T3/el unknown
-
1999
- 1999-11-29 FI FI992546A patent/FI107265B/sv not_active IP Right Cessation
Also Published As
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| FI107265B (sv) | Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster | |
| FI95929B (sv) | Framställning av hepatitis B ytantigen i jäst | |
| KR940005588B1 (ko) | 효모에서 b형 간염 바이러스 단백질을 제조하는 방법 | |
| FI94427B (sv) | Regleringsregion för heterologisk genexpression i jäst | |
| KR0181940B1 (ko) | 변형된 HBsAg 라아지 단백질 | |
| USRE35749E (en) | Constructs of glyceraldehyde-3-phosphate dehydrogenase promoter and methods for expressing genes using said constructs | |
| US5089398A (en) | Enhanced yeast transcription employing hybrid GAPDH promoter region constructs | |
| DK169152B1 (da) | Fremgangsmåde til fremstilling af et antistof mod HBsAg | |
| AU605036B2 (en) | Expression of hepatitis B2 pres protein in methylothrophic yeasts | |
| AU619753B2 (en) | Hepatitis b surface antigen vaccine | |
| EP0301083A1 (en) | Retroviral expression vectors and methods for producing hbv antigens | |
| JPH06172392A (ja) | 粒子を形成する多重ｂ型肝炎ウイルス表層タンパク質 | |
| EP0235430A1 (en) | Novel protein and production thereof | |
| PL168408B1 (pl) | Sposób wytwarzania stransformowanego mikroorganizmu PL | |
| KR900005534B1 (ko) | 복합 프로모터를 이용한 효모 발현 벡터에 의한 b형 간염 바이러스 표면항원들의 제조방법 | |
| EP0277770A1 (en) | Alpha-mating factor promoter is modified by deleting pre-pro secretory leader sequence | |
| IE903178L (en) | Synthesis of human virus antigens by yeast |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MA | Patent expired |