DK2870241T3 - Sammensætning udvisende aktiveringsinduceret cytidindeaminase (aid)-aktivitet, fremgangsmåde til tilvejebringelse af denne sammensætning i et bakterielt system samt anvendelse af sammensætningen ved analyse af dna/rna-aminering/deaminering og/eller -methylering/demethylering - Google Patents
Sammensætning udvisende aktiveringsinduceret cytidindeaminase (aid)-aktivitet, fremgangsmåde til tilvejebringelse af denne sammensætning i et bakterielt system samt anvendelse af sammensætningen ved analyse af dna/rna-aminering/deaminering og/eller -methylering/demethylering Download PDFInfo
- Publication number
- DK2870241T3 DK2870241T3 DK14703938.2T DK14703938T DK2870241T3 DK 2870241 T3 DK2870241 T3 DK 2870241T3 DK 14703938 T DK14703938 T DK 14703938T DK 2870241 T3 DK2870241 T3 DK 2870241T3
- Authority
- DK
- Denmark
- Prior art keywords
- aid
- composition
- activity
- gst
- leu
- Prior art date
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y305/00—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
- C12Y305/04—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in cyclic amidines (3.5.4)
- C12Y305/04005—Cytidine deaminase (3.5.4.5)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
- C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
- C07K2319/23—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a GST-tag
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/50—Fusion polypeptide containing protease site
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Claims (4)
1. Sammensætning, der udviser human aktiverings-induceret cytidindeaminase (AID) ved methyleret og/eller ikke-methyleret cytidin, kendetegnet ved, at den indeholder en protein, der fremstilles som følge af expression af modificeret gen af humant AID i bakterieceller, og at det modificerede gen omfatter: (i) den komplette cDNA-sekvens, som koder den humane ATP med STOP-codons, bort set fra 5' og 3' ikke-translaterede regioner, inklusive positioner 80-679 af SEQ. ID No 1; og (ii) sekvenser, som koder to tags: GST og hexahistidin; og (iii) sekvenser, som koder to af proteaseme thrombin og enterokinase erkendte motiver; og (iv) sekvenser, som koder bestemte elementer af fusionsproteiner i følgende rækkefølge (fra ende 5'- til 3'): GST-tag, et af thrombin genkendt motiv, hexahistidin-tag, et motiv genkendt af enterokinase, AID; altsammen udgørende sekvensen SEQ. ID No 3 eller sekvenserne udvisende i det mindste 85 % aminosyrelighed med SEQ. ID No 4; hvor den bakterielle cellekultur befinder sig i vækst i et medium, som indeholder en kilde med zink-ioner, og at den bakterielle cellekultur befinder sig i vækst ved en temperatur fra 15 til 30 °C, fortrinsvis 18 °C.
2. Fremgangsmåde til i et bakterielt system at tilvejebringe en sammensætning, der udviser et modificeret gen af humant ATP i bakterielle celler, hvor det modificerede gen omfatter: human aktiverings-induceret cytidindeaminase (AID) ved methyleret og/eller ikke-methyleret cytidin, og fremgangsmåden omfatter en expression af: (i) den komplette cDNA-sekvens, som koder den humane ATP med STOP-codons, bort set fra 5' og 3' ikke-translaterede regioner, inklusive positioner 80-679 af SEQ. ID No 1; og (ii) sekvenser, som koder to tags: GST og hexahistidin; og (iii) sekvenser, som koder to motiver genkendt af proteaserne: thrombin og enterokinase; og (iv) sekvenser, som koder bestemte elementer af fusionsproteiner i følgende rækkefølge (fra ende 5'- til 3'): GST-tag, et af thrombin genkendt motiv, hexahistidin-tag, et motiv genkendt af enterokinase, AID; alt sammen udgørende sekvensen SEQ. ID No 3 eller sekvenserne udvisende i det mindste 85 % aminosyrelighed med SEQ. ID No 4; og hvor den bakterielle cellekultur foreligger i vækst i et medium, som indeholder en kilde med zink-ioner, og at den bakterielle cellekultur forligger i vækst ved en temperatur fra 15 til 30 °C, fortrinsvis 18 °C.
3. Fremgangsmåde ifølge krav 2 kendetegnet ved ,at zinkion-kilden er zinkchlorid med koncentration fra 0,01 til mM, fortrinsvis 0,06 mM.
4. Anvendelse af sammensætningen ifølge krav 1 ved analyser af DNA/RNA-aminering/-deaminering og -methylering/-demethylering.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PL402440A PL225333B1 (pl) | 2013-01-14 | 2013-01-14 | Preparat wykazujący aktywność ludzkiej deaminazy cytydyny indukowanej aktywacją limfocytów B (AID) oraz sposób otrzymywania tego preparatu w systemie bakteryjnym |
| PCT/PL2014/000003 WO2014109656A1 (en) | 2013-01-14 | 2014-01-14 | Modified gene of human activation-induced cytidine deaminase (aid), a method of modification of human aid gene, a composition showing aid activity, a method of preparation of such composition in a bacterial system and use of the composition in the analyses of dna/rna amination/deamination and/or methylation/demethylation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2870241T3 true DK2870241T3 (da) | 2018-06-06 |
Family
ID=50073407
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK14703938.2T DK2870241T3 (da) | 2013-01-14 | 2014-01-14 | Sammensætning udvisende aktiveringsinduceret cytidindeaminase (aid)-aktivitet, fremgangsmåde til tilvejebringelse af denne sammensætning i et bakterielt system samt anvendelse af sammensætningen ved analyse af dna/rna-aminering/deaminering og/eller -methylering/demethylering |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US20150044675A1 (da) |
| EP (1) | EP2870241B1 (da) |
| DK (1) | DK2870241T3 (da) |
| PL (1) | PL225333B1 (da) |
| WO (1) | WO2014109656A1 (da) |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0851929A1 (en) * | 1995-07-31 | 1998-07-08 | Centre De Recherche De L'hopital Ste-Justine | CYTIDINE DEAMINASE cDNA AS A POSITIVE SELECTABLE MARKER FOR GENE TRANSFER, GENE THERAPY AND PROTEIN SYNTHESIS |
| EP1575992A4 (en) * | 2002-08-05 | 2007-02-21 | Univ Rochester | CHIMERIC PROTEINS WITH PROTEIN TRANSDUCTION FIELD / DOMAINE DESAMINASE, ASSOCIATED COMPOUNDS AND CORRESPONDING USES |
| CA2757178C (en) * | 2009-04-03 | 2020-05-19 | Medical Research Council | Mutants of activation-induced cytidine deaminase (aid) and methods of use |
-
2013
- 2013-01-14 PL PL402440A patent/PL225333B1/pl unknown
-
2014
- 2014-01-14 DK DK14703938.2T patent/DK2870241T3/da active
- 2014-01-14 WO PCT/PL2014/000003 patent/WO2014109656A1/en active Application Filing
- 2014-01-14 EP EP14703938.2A patent/EP2870241B1/en active Active
- 2014-01-14 US US14/384,665 patent/US20150044675A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| PL402440A1 (pl) | 2014-07-21 |
| WO2014109656A1 (en) | 2014-07-17 |
| US20150044675A1 (en) | 2015-02-12 |
| PL225333B1 (pl) | 2017-03-31 |
| EP2870241B1 (en) | 2018-03-14 |
| EP2870241A1 (en) | 2015-05-13 |
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