CN111094572A - 一种密码子优化的人胰岛素类似物前体基因和信号肽基因 - Google Patents
一种密码子优化的人胰岛素类似物前体基因和信号肽基因 Download PDFInfo
- Publication number
- CN111094572A CN111094572A CN201980004477.8A CN201980004477A CN111094572A CN 111094572 A CN111094572 A CN 111094572A CN 201980004477 A CN201980004477 A CN 201980004477A CN 111094572 A CN111094572 A CN 111094572A
- Authority
- CN
- China
- Prior art keywords
- nucleic acid
- acid molecule
- seq
- human insulin
- molecule encoding
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000002243 precursor Substances 0.000 title claims abstract description 66
- 108010076504 Protein Sorting Signals Proteins 0.000 title claims abstract description 27
- PBGKTOXHQIOBKM-FHFVDXKLSA-N insulin (human) Chemical class C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]1CSSC[C@H]2C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3C=CC(O)=CC=3)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3NC=NC=3)NC(=O)[C@H](CO)NC(=O)CNC1=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O)=O)CSSC[C@@H](C(N2)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](NC(=O)CN)[C@@H](C)CC)[C@@H](C)CC)[C@@H](C)O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C=CC=CC=1)C(C)C)C1=CN=CN1 PBGKTOXHQIOBKM-FHFVDXKLSA-N 0.000 title claims description 63
- 108090000623 proteins and genes Proteins 0.000 title description 33
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 94
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 91
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 91
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 9
- 239000013598 vector Substances 0.000 claims description 25
- 101000976075 Homo sapiens Insulin Proteins 0.000 claims description 17
- 239000013604 expression vector Substances 0.000 claims description 16
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 15
- 241000235058 Komagataella pastoris Species 0.000 claims description 14
- 150000001413 amino acids Chemical group 0.000 claims description 13
- 229920001184 polypeptide Polymers 0.000 claims description 13
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 13
- 238000001976 enzyme digestion Methods 0.000 claims description 11
- 101800001707 Spacer peptide Proteins 0.000 claims description 10
- 238000003776 cleavage reaction Methods 0.000 claims description 10
- 230000007017 scission Effects 0.000 claims description 10
- 238000012545 processing Methods 0.000 claims description 9
- 101000702488 Rattus norvegicus High affinity cationic amino acid transporter 1 Proteins 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 5
- 239000004472 Lysine Substances 0.000 claims description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 4
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 claims description 4
- 238000006467 substitution reaction Methods 0.000 claims 2
- 238000002360 preparation method Methods 0.000 claims 1
- 230000009465 prokaryotic expression Effects 0.000 claims 1
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical class N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 abstract description 76
- 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 abstract description 18
- 239000004026 insulin derivative Substances 0.000 abstract description 3
- 230000028327 secretion Effects 0.000 abstract description 2
- 102000004877 Insulin Human genes 0.000 description 36
- 108090001061 Insulin Proteins 0.000 description 36
- 229940125396 insulin Drugs 0.000 description 36
- 230000014509 gene expression Effects 0.000 description 23
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 241000894006 Bacteria Species 0.000 description 15
- 210000004027 cell Anatomy 0.000 description 15
- 108020004705 Codon Proteins 0.000 description 9
- 238000004519 manufacturing process Methods 0.000 description 8
- 238000003259 recombinant expression Methods 0.000 description 8
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 7
- 239000012634 fragment Substances 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 238000005457 optimization Methods 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 230000003248 secreting effect Effects 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 238000012795 verification Methods 0.000 description 4
- 241001506991 Komagataella phaffii GS115 Species 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 210000003527 eukaryotic cell Anatomy 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000009776 industrial production Methods 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 108010025188 Alcohol oxidase Proteins 0.000 description 2
- 102100036826 Aldehyde oxidase Human genes 0.000 description 2
- 108010075254 C-Peptide Proteins 0.000 description 2
- 108700010070 Codon Usage Proteins 0.000 description 2
- 102000004533 Endonucleases Human genes 0.000 description 2
- 108010042407 Endonucleases Proteins 0.000 description 2
- 101000928314 Homo sapiens Aldehyde oxidase Proteins 0.000 description 2
- 102000003960 Ligases Human genes 0.000 description 2
- 108090000364 Ligases Proteins 0.000 description 2
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 2
- 239000004473 Threonine Substances 0.000 description 2
- 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
- 229960000723 ampicillin Drugs 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 210000003000 inclusion body Anatomy 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 150000004668 long chain fatty acids Chemical group 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 230000006798 recombination Effects 0.000 description 2
- 238000005215 recombination Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- 239000007222 ypd medium Substances 0.000 description 2
- 101150084750 1 gene Proteins 0.000 description 1
- VOUAQYXWVJDEQY-QENPJCQMSA-N 33017-11-7 Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)NCC(=O)NCC(=O)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N1[C@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)CCC1 VOUAQYXWVJDEQY-QENPJCQMSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 208000013824 Acidemia Diseases 0.000 description 1
- 208000010444 Acidosis Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 101500025353 Homo sapiens Insulin A chain Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 208000007976 Ketosis Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- -1 antibodies Proteins 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000023852 carbohydrate metabolic process Effects 0.000 description 1
- 235000021256 carbohydrate metabolism Nutrition 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- 229940047122 interleukins Drugs 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 230000004140 ketosis Effects 0.000 description 1
- 230000004130 lipolysis Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 125000001419 myristoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 230000000153 supplemental effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000013603 viral vector Substances 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
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/62—Insulins
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/50—Fusion polypeptide containing protease site
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/84—Pichia
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Diabetes (AREA)
- Endocrinology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
提供一种密码子优化的人胰岛素类似物前体基因和信号肽基因的核酸分子。该核酸分子包含一种融合胰岛素类似物前体的编码核酸分子和一种酵母分泌信号肽α‑因子的编码核酸分子。该核酸分子使得胰岛素类似物前体在毕赤酵母中的表达量提高，降低人胰岛素类似物的生产成本。
Description
PCT国内申请，说明书已公开。
Claims (13)
- PCT国内申请，权利要求书已公开。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2018101317943 | 2018-02-09 | ||
| CN201810131794 | 2018-02-09 | ||
| PCT/CN2019/074384 WO2019154311A1 (zh) | 2018-02-09 | 2019-02-01 | 一种密码子优化的人胰岛素类似物前体基因和信号肽基因 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111094572A true CN111094572A (zh) | 2020-05-01 |
| CN111094572B CN111094572B (zh) | 2023-04-04 |
Family
ID=67548783
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201980004477.8A Active CN111094572B (zh) | 2018-02-09 | 2019-02-01 | 一种密码子优化的人胰岛素类似物前体基因和信号肽基因 |
Country Status (12)
| Country | Link |
|---|---|
| US (1) | US20210032307A1 (zh) |
| EP (1) | EP3750998A4 (zh) |
| JP (1) | JP2021513330A (zh) |
| KR (1) | KR20200119237A (zh) |
| CN (1) | CN111094572B (zh) |
| AU (1) | AU2019218315A1 (zh) |
| BR (1) | BR112020015238A2 (zh) |
| CA (1) | CA3086618A1 (zh) |
| MX (1) | MX2020007929A (zh) |
| RU (1) | RU2020128190A (zh) |
| TW (1) | TW201934752A (zh) |
| WO (1) | WO2019154311A1 (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115927360A (zh) * | 2020-05-26 | 2023-04-07 | 北京惠之衡生物科技有限公司 | 密码子优化的门冬胰岛素前体基因、重组载体、基因工程菌及其应用 |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117384275B (zh) * | 2023-12-13 | 2024-03-15 | 北京科为博生物科技有限公司 | ***突变体igf1m及其应用 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998028429A1 (en) * | 1996-12-20 | 1998-07-02 | Novo Nordisk A/S | N-terminally extended proteins expressed in yeast |
| CN1873006A (zh) * | 2005-05-30 | 2006-12-06 | 上海新生源医药研究有限公司 | 一种重组人胰岛素原的生产方法 |
| CN102439154A (zh) * | 2009-03-12 | 2012-05-02 | 比格科技私人有限公司 | 一种多核苷酸和多肽链序列及其方法 |
| CN103889444A (zh) * | 2011-08-08 | 2014-06-25 | 默沙东公司 | N-糖基化的胰岛素类似物 |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA1340772C (en) * | 1987-12-30 | 1999-09-28 | Patricia Tekamp-Olson | Expression and secretion of heterologous protiens in yeast employing truncated alpha-factor leader sequences |
| UA45321C2 (uk) | 1993-09-17 | 2002-04-15 | Ново Нордіск А/С | Похідне інсуліну, фармацевтична композиція для лікування діабету та спосіб лікування діабету |
| US5639642A (en) * | 1994-06-16 | 1997-06-17 | Novo Nordisk A/S | Synthetic leader peptide sequences |
| CN1302333A (zh) * | 1998-01-23 | 2001-07-04 | 诺沃挪第克公司 | 在酵母中制备目的多肽的方法 |
| AR025646A1 (es) * | 2000-09-13 | 2002-12-04 | Beta Lab Sa | Cepa de levaduras metilotroficas recombinantes productoras de un precursor de insulina, construcciones de adn y metodo para obtener la cepa. |
| ES2313018T3 (es) * | 2003-06-17 | 2009-03-01 | Sembiosys Genetics Inc. | Procedimiento de produccion de insulina en plantas. |
| WO2005012347A2 (en) | 2003-08-05 | 2005-02-10 | Novo Nordisk A/S | Novel insulin derivatives |
| AU2012350586B2 (en) * | 2011-12-15 | 2017-02-02 | Jiangsu Hengrui Medicine Co., Ltd. | Human insulin analogue and acylated derivative thereof |
| US10815287B2 (en) | 2016-08-02 | 2020-10-27 | Jiangsu Hengrui Medicine Co., Ltd. | Acylated derivative of human insulin or analogue thereof |
-
2019
- 2019-02-01 BR BR112020015238-2A patent/BR112020015238A2/pt not_active IP Right Cessation
- 2019-02-01 EP EP19751736.0A patent/EP3750998A4/en not_active Withdrawn
- 2019-02-01 CA CA3086618A patent/CA3086618A1/en active Pending
- 2019-02-01 WO PCT/CN2019/074384 patent/WO2019154311A1/zh unknown
- 2019-02-01 RU RU2020128190A patent/RU2020128190A/ru unknown
- 2019-02-01 AU AU2019218315A patent/AU2019218315A1/en not_active Abandoned
- 2019-02-01 MX MX2020007929A patent/MX2020007929A/es unknown
- 2019-02-01 US US16/967,117 patent/US20210032307A1/en not_active Abandoned
- 2019-02-01 KR KR1020207020915A patent/KR20200119237A/ko not_active Application Discontinuation
- 2019-02-01 TW TW108104187A patent/TW201934752A/zh unknown
- 2019-02-01 CN CN201980004477.8A patent/CN111094572B/zh active Active
- 2019-02-01 JP JP2020541883A patent/JP2021513330A/ja not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998028429A1 (en) * | 1996-12-20 | 1998-07-02 | Novo Nordisk A/S | N-terminally extended proteins expressed in yeast |
| CN1873006A (zh) * | 2005-05-30 | 2006-12-06 | 上海新生源医药研究有限公司 | 一种重组人胰岛素原的生产方法 |
| CN102439154A (zh) * | 2009-03-12 | 2012-05-02 | 比格科技私人有限公司 | 一种多核苷酸和多肽链序列及其方法 |
| CN103889444A (zh) * | 2011-08-08 | 2014-06-25 | 默沙东公司 | N-糖基化的胰岛素类似物 |
Non-Patent Citations (1)
| Title |
|---|
| 陈频等: "《全科医师糖尿病临床诊疗手册》", 30 November 2017, 福建科学技术出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115927360A (zh) * | 2020-05-26 | 2023-04-07 | 北京惠之衡生物科技有限公司 | 密码子优化的门冬胰岛素前体基因、重组载体、基因工程菌及其应用 |
| CN115927360B (zh) * | 2020-05-26 | 2023-07-14 | 北京惠之衡生物科技有限公司 | 密码子优化的门冬胰岛素前体基因、重组载体、基因工程菌及其应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CA3086618A1 (en) | 2019-08-15 |
| TW201934752A (zh) | 2019-09-01 |
| RU2020128190A (ru) | 2022-03-09 |
| AU2019218315A1 (en) | 2020-07-09 |
| MX2020007929A (es) | 2020-10-01 |
| JP2021513330A (ja) | 2021-05-27 |
| US20210032307A1 (en) | 2021-02-04 |
| RU2020128190A3 (zh) | 2022-03-09 |
| CN111094572B (zh) | 2023-04-04 |
| BR112020015238A2 (pt) | 2020-12-29 |
| WO2019154311A1 (zh) | 2019-08-15 |
| EP3750998A4 (en) | 2022-02-23 |
| EP3750998A1 (en) | 2020-12-16 |
| KR20200119237A (ko) | 2020-10-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111094572B (zh) | 一种密码子优化的人胰岛素类似物前体基因和信号肽基因 | |
| CN109295067B (zh) | 一种密码子优化的德谷胰岛素前体基因及其表达方法 | |
| CN108060195A (zh) | 一种发酵生产重组胰高血糖素样肽-1蛋白的方法 | |
| CN111944823B (zh) | 适合于酵母表达的抗菌肽Hepcidin优化基因及其表达载体和应用 | |
| EP1185675B1 (en) | Escherichia coli strain secreting human granulocyte colony stimulating factor (g-csf) | |
| CN102016043A (zh) | 用新融合配偶体生产重组蛋白的方法 | |
| EP0643133A2 (de) | Rekombinante D-Hydantoinase, Verfahren zur Herstellung und Verwendung | |
| JP6943841B2 (ja) | メタノール資化性酵母由来新規タンパク質及びそれを用いた目的タンパク質の製造方法 | |
| CN110592084A (zh) | 一种rhtA基因启动子改造的重组菌株及其构建方法与应用 | |
| CN106632654B (zh) | 一种优化的pIFN-γ肽链及在提高毕赤酵母分泌表达猪IFN-γ产量和活性中的应用 | |
| JP7012663B2 (ja) | 新規宿主細胞及びそれを用いた目的タンパク質の製造方法 | |
| JP2021101733A (ja) | 新規宿主細胞及びそれを用いた目的タンパク質の製造方法 | |
| CN113462713B (zh) | 提高胰高血糖素样肽六连体在毕赤酵母中表达水平的方法 | |
| CN101503679A (zh) | Qsox及其突变体蛋白及制备方法和应用 | |
| US11866714B2 (en) | Promoter for yeast | |
| CA1283068C (en) | Plasmid for expression of penicillin g amidase | |
| JP2020513834A (ja) | ペプチドの発現および大規模生産 | |
| AU621277B2 (en) | Shortened phosphoglycerate kinase promoter | |
| CN115960920B (zh) | 协氧蛋白FHb及其重组菌X33-pPICZαA-102C300C-FHb2和应用 | |
| US7354994B2 (en) | Recombinant IGF expression systems | |
| RU2263147C1 (ru) | РЕКОМБИНАНТНАЯ ПЛАЗМИДНАЯ ДНК pHINS05, КОДИРУЮЩАЯ ГИБРИДНЫЙ ПОЛИПЕПТИД, СОДЕРЖАЩИЙ ПРОИНСУЛИН ЧЕЛОВЕКА, КЛЕТКА ESCHERICHIA COLI, ТРАНСФОРМИРОВАННАЯ РЕКОМБИНАНТНОЙ ПЛАЗМИДНОЙ ДНК pHINS05, И ШТАММ БАКТЕРИЙ ESCHERICHIA COLI JM109/pHINS05 - ПРОДУЦЕНТ ГИБРИДНОГО ПОЛИПЕПТИДА, СОДЕРЖАЩЕГО ПРОИНСУЛИН ЧЕЛОВЕКА | |
| US10273520B2 (en) | Means and methods for hyper-production of authentic human basic fibroblast growth factor in Escherichia coli | |
| RU2515061C1 (ru) | РЕКОМБИНАТНАЯ ПЛАЗМИДНАЯ ДНК рНI03 КОДИРУЮЩАЯ, ГИБРИДНЫЙ БЕЛОК С ПРОИНСУЛИНОМ ЧЕЛОВЕКА, КЛЕТКА Escherichia coli, ТРАНСФОРМИРОВАННАЯ РЕКОМБИНАНТНОЙ ПЛАЗМИДНОЙ ДНК рНI03, И ШТАММ БАКТЕРИЙ Escherichia coli JM109/pHI03-ПРОДУЦЕНТ ГИБРИДНОГО БЕЛКА-ПРЕДШЕСТВЕННИКА ИНСУЛИНА ЧЕЛОВЕКА | |
| CN104672334B (zh) | 重组长链人***-i的制备方法 | |
| CN117417842A (zh) | 一种高效表达猪胰蛋白酶重组毕赤酵母菌株及其应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 40025349 Country of ref document: HK |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |