BRPI0706146A2 - production of bos taurus growth hormone (gh) by pichia pastoris yeast using a synthetic gene sequence - Google Patents
production of bos taurus growth hormone (gh) by pichia pastoris yeast using a synthetic gene sequenceInfo
- Publication number
- BRPI0706146A2 BRPI0706146A2 BRPI0706146A BRPI0706146A2 BR PI0706146 A2 BRPI0706146 A2 BR PI0706146A2 BR PI0706146 A BRPI0706146 A BR PI0706146A BR PI0706146 A2 BRPI0706146 A2 BR PI0706146A2
- Authority
- BR
- Brazil
- Prior art keywords
- expression
- pastoris
- growth hormone
- sequence
- yeast
- Prior art date
Links
- 241000235058 Komagataella pastoris Species 0.000 title abstract 8
- 240000004808 Saccharomyces cerevisiae Species 0.000 title abstract 5
- 238000004519 manufacturing process Methods 0.000 title abstract 4
- 108700005078 Synthetic Genes Proteins 0.000 title abstract 3
- 101000824887 Bos taurus Somatotropin Proteins 0.000 title abstract 2
- 108020004705 Codon Proteins 0.000 abstract 3
- 108010051696 Growth Hormone Proteins 0.000 abstract 3
- 102000018997 Growth Hormone Human genes 0.000 abstract 3
- 239000002773 nucleotide Substances 0.000 abstract 3
- 125000003729 nucleotide group Chemical group 0.000 abstract 3
- 239000013604 expression vector Substances 0.000 abstract 2
- 239000000122 growth hormone Substances 0.000 abstract 2
- 239000005556 hormone Substances 0.000 abstract 2
- 229940088597 hormone Drugs 0.000 abstract 2
- 108090000623 proteins and genes Proteins 0.000 abstract 2
- 241000283690 Bos taurus Species 0.000 abstract 1
- 101001075376 Bos taurus Gamma-glutamyl hydrolase Proteins 0.000 abstract 1
- 108091026890 Coding region Proteins 0.000 abstract 1
- 230000006978 adaptation Effects 0.000 abstract 1
- 125000000539 amino acid group Chemical group 0.000 abstract 1
- 230000015572 biosynthetic process Effects 0.000 abstract 1
- 108010006025 bovine growth hormone Proteins 0.000 abstract 1
- 238000004113 cell culture Methods 0.000 abstract 1
- 238000010367 cloning Methods 0.000 abstract 1
- 239000002299 complementary DNA Substances 0.000 abstract 1
- 238000000326 densiometry Methods 0.000 abstract 1
- 238000011156 evaluation Methods 0.000 abstract 1
- 238000012215 gene cloning Methods 0.000 abstract 1
- 238000000034 method Methods 0.000 abstract 1
- 238000005457 optimization Methods 0.000 abstract 1
- 238000011002 quantification Methods 0.000 abstract 1
- 239000013606 secretion vector Substances 0.000 abstract 1
- 238000003786 synthesis reaction Methods 0.000 abstract 1
- 230000009466 transformation Effects 0.000 abstract 1
- 238000011144 upstream manufacturing Methods 0.000 abstract 1
- 210000005253 yeast cell Anatomy 0.000 abstract 1
Classifications
-
- 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
- C07K14/61—Growth hormone [GH], i.e. somatotropin
-
- 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
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)
- Bioinformatics & Cheminformatics (AREA)
- Endocrinology (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Mycology (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
PRODUçãO DO HORMÈNIO DE CRESCIMENTO (GH) DE BOS TAURUS PELA LEVEDURA PICHIA PASTORIS UTILIZANDO UMA SEQüêNCIA GéNICA SINTéTICA. O objeto da presente invenção trata da produção do hormónio de crescimento bovino recombinante (rbGH) utilizando como sistema de expressão a levedura P. pastoris. A inovação está focada na utilização de uma sequência gênica sintética que utiliza códons preferenciais e conteúdo otimizado de bases nucleotídicas GC para P. pastoris. O processo de produção do bGH recombinante utilizando como sistema de expressão a levedura P. pastoris compreende: (a) síntese do cDNA codificador do GH bovino (Bos taurus taurus) utilizando como referência as sequências gênicas descritas no GenBank (gi:2168581, gi:2168481, gi:2168363) considerando as seguintes alteraçóes: (1) Adaptação da seqúência sintetizada do referido hormónio para os códons preferenciais da levedura P. pastoris e otimização do conteúdo de bases nucleotídica GO; (II> Inclusão de sítio de restrição Xhol na extremidade 5', e de Xbal, na extremidade 3', com a finalidade de facilitar o processo de clonagem gênica em vetor de expressão; (III) Adição de sequência de nucleotídeos que codifiquem para 6 resíduos de aminoácidos na extremidade 5' imediatamente upstream da seqúência codante do bGH, e adição de um stop codon downstream à sequência; (b) Clonagem do gene sintetizado em vetor de expressão/secreção para P. pastoris (pPICZ<244>A) utilizando metodologias de biologia molecular; (c) Transformação da linhagem de P. pastoris GSII5 para expressão extracelular de bGH; (d) Cultura celular de levedura em frascos aletados de 1 L para expressão do hormónio de crescimento; (e) Avaliação da expressão do hormónio de crescimento através de Western Blotting; (f) Quantificação da eficiência de expressão do hormónio de crescimento recombinante por teste de Bradford e densitometria de gel.BOS TAURUS GROWTH HORMONE (GH) PRODUCTION BY PICHIA PASTORIS Yeast USING A SYNTHETIC GENE SEQUENCE. The object of the present invention is the production of recombinant bovine growth hormone (rbGH) using as expression system P. pastoris yeast. Innovation is focused on the use of a synthetic gene sequence that utilizes preferred codons and optimized GC nucleotide base content for P. pastoris. The production process of recombinant bGH using P. pastoris yeast as an expression system comprises: (a) synthesis of bovine GH coding cDNA (Bos taurus taurus) using as reference the gene sequences described in GenBank (gi: 2168581, gi: 2168481, gi: 2168363) considering the following changes: (1) Adaptation of the synthesized sequence of said hormone to P. pastoris preferential yeast codons and optimization of GO nucleotide base content; (II> Inclusion of Xhol restriction site at the 5 'end, and Xbal restriction at the 3' end, to facilitate the expression vector gene cloning process; (III) Addition of 6-encoding nucleotide sequence amino acid residues at the 5 'end immediately upstream of the bGH coding sequence, and the addition of a downstream stop codon to the sequence (b) cloning of the expression / secretion vector gene synthesized for P. pastoris (pPICZ <244> A) using (c) Transformation of P. pastoris GSII5 strain to bGH extracellular expression (d) Yeast cell culture in 1 L finned flasks for growth hormone expression (e) Evaluation of hormone expression (f) Quantification of recombinant growth hormone expression efficiency by Bradford test and gel densitometry.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| BRPI0706146 BRPI0706146A2 (en) | 2007-12-14 | 2007-12-14 | production of bos taurus growth hormone (gh) by pichia pastoris yeast using a synthetic gene sequence |
| PCT/BR2008/000385 WO2009076737A1 (en) | 2007-12-14 | 2008-12-12 | Production of recombinant bos taurus growth hormone with pichia pastoris |
| CA2746905A CA2746905A1 (en) | 2007-12-14 | 2008-12-12 | Production of recombinant bos taurus growth hormone with pichia pastoris |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| BRPI0706146 BRPI0706146A2 (en) | 2007-12-14 | 2007-12-14 | production of bos taurus growth hormone (gh) by pichia pastoris yeast using a synthetic gene sequence |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| BRPI0706146A2 true BRPI0706146A2 (en) | 2009-08-18 |
Family
ID=40795130
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| BRPI0706146 BRPI0706146A2 (en) | 2007-12-14 | 2007-12-14 | production of bos taurus growth hormone (gh) by pichia pastoris yeast using a synthetic gene sequence |
Country Status (3)
| Country | Link |
|---|---|
| BR (1) | BRPI0706146A2 (en) |
| CA (1) | CA2746905A1 (en) |
| WO (1) | WO2009076737A1 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102899331B (en) * | 2012-09-25 | 2015-04-22 | 广东省农业科学院兽医研究所 | Complex duck interferon-alpha gene, and recombinant vector and application thereof |
| CN112175964B (en) * | 2020-10-23 | 2022-03-29 | 安徽中起生物科技有限公司 | Recombinant pig growth hormone and preparation method and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2002218893A1 (en) * | 2001-12-21 | 2003-07-09 | Thromb-X Nv | Compositions for the in vitro derivation and culture of embryonic stem (es) cell lines with germline transmission capability |
| CN100482796C (en) * | 2003-10-17 | 2009-04-29 | 任宏伟 | Expression of recombinant ox growth hormone in pichia pastoris |
-
2007
- 2007-12-14 BR BRPI0706146 patent/BRPI0706146A2/en not_active Application Discontinuation
-
2008
- 2008-12-12 CA CA2746905A patent/CA2746905A1/en not_active Abandoned
- 2008-12-12 WO PCT/BR2008/000385 patent/WO2009076737A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2009076737A1 (en) | 2009-06-25 |
| CA2746905A1 (en) | 2009-12-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | High-level expression and purification of soluble recombinant FGF21 protein by SUMO fusion in Escherichia coli | |
| Shi et al. | Latent TGF-β structure and activation | |
| HN2009001593A (en) | GLUCAGON / GLP-1 RECEIVER COAGNISTS | |
| Rosenfeld et al. | Biochemical, biophysical, and pharmacological characterization of bacterially expressed human agouti-related protein | |
| RU2012106148A (en) | OPTIONAL FORMS OF URATOXIDASE AND THEIR APPLICATION | |
| KR950000301B1 (en) | Gm-csf protein its derivatives preparation of such proteins and theiruse | |
| DK1987150T3 (en) | Selection of host cells expressing high-level protein | |
| BRPI0706146A2 (en) | production of bos taurus growth hormone (gh) by pichia pastoris yeast using a synthetic gene sequence | |
| Su et al. | High-level expression and purification of human epidermal growth factor with SUMO fusion in Escherichia coli | |
| He et al. | Nanoluciferase as a novel quantitative protein fusion tag: Application for overexpression and bioluminescent receptor-binding assays of human leukemia inhibitory factor | |
| Shabanpoor et al. | Design and development of analogues of dimers of insulin‐like peptide 3 B‐chain as high‐affinity antagonists of the RXFP2 receptor | |
| Wu et al. | Application of the novel bioluminescent ligand–receptor binding assay to relaxin-RXFP1 system for interaction studies | |
| Moritz et al. | Determination of the Disulfide Structure andN-Glycosylation Sites of the Extracellular Domain of the Human Signal Transducer gp130 | |
| CN101760470B (en) | Method for preparing human growth hormone recombinant | |
| CN105829350A (en) | Enterokinase cleavable polypeptides | |
| Rösch et al. | The topogenic contribution of uncharged amino acids on signal sequence orientation in the endoplasmic reticulum | |
| McNicoll et al. | Localization by photoaffinity labeling of natriuretic peptide receptor-A binding domain | |
| CN101281201A (en) | Method using implicit peptides trans splicing to manufacture protein molecular weight standard | |
| Hammond et al. | Molecular cloning and expression of leptin in gray and harbor seal blubber, bone marrow, and lung and its potential role in marine mammal respiratory physiology | |
| Klose et al. | Impact of N-Terminal Domains for Corticotropin-Releasing Factor (CRF) Receptor− Ligand Interactions | |
| Mullin et al. | Improved secretion of recombinant human IL-25 in HEK293 cells using a signal peptide-pro-peptide domain derived from Trypsin-1 | |
| Zou et al. | An improved recombinant mammalian cell expression system for human transforming growth factor-β2 and-β3 preparations | |
| BRPI0715331B8 (en) | polypeptide comprising an n-terminal portion and a c-terminal portion, and use thereof | |
| Speck et al. | Expression and characterization of relaxin family peptide receptor 1 variants | |
| Bals et al. | Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| B03A | Publication of an application: publication of a patent application or of a certificate of addition of invention | ||
| B11K | Dismissal: dismissal - article 17, par 2 of industrial property law |
Free format text: PRIORIDADE INTERNA DO PI0806654-0. |
|
| B03G | Publication of an application: publication cancelled |
Free format text: REFERENTE A RPI 2015 DE 18/08/2009. |
|
| B03F | Publication of an application: publication of application definitely dismissed - article 216, par 2 and article 17, par 2 of industr. prop. law |