EP0847444A1 - Genetische transformation von ciliatenzellen durch microcarrier- bombardement mit dna-beladenen goldpartikeln - Google Patents
Genetische transformation von ciliatenzellen durch microcarrier- bombardement mit dna-beladenen goldpartikelnInfo
- Publication number
- EP0847444A1 EP0847444A1 EP97930486A EP97930486A EP0847444A1 EP 0847444 A1 EP0847444 A1 EP 0847444A1 EP 97930486 A EP97930486 A EP 97930486A EP 97930486 A EP97930486 A EP 97930486A EP 0847444 A1 EP0847444 A1 EP 0847444A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- dna
- ciliate
- heterologous dna
- cells
- origin
- 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.)
- Withdrawn
Links
- 241000223782 Ciliophora Species 0.000 title claims abstract description 71
- 239000002245 particle Substances 0.000 title claims abstract description 13
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 title claims abstract description 9
- 239000010931 gold Substances 0.000 title claims abstract description 9
- 229910052737 gold Inorganic materials 0.000 title claims abstract description 9
- 230000009466 transformation Effects 0.000 title claims description 25
- 230000002068 genetic effect Effects 0.000 title description 2
- 238000000034 method Methods 0.000 claims abstract description 57
- 108020004414 DNA Proteins 0.000 claims description 42
- 108090000623 proteins and genes Proteins 0.000 claims description 20
- 239000013604 expression vector Substances 0.000 claims description 12
- 239000013612 plasmid Substances 0.000 claims description 10
- 238000013518 transcription Methods 0.000 claims description 9
- 230000035897 transcription Effects 0.000 claims description 9
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 108091026890 Coding region Proteins 0.000 claims description 6
- 241000701489 Cauliflower mosaic virus Species 0.000 claims description 5
- 230000008488 polyadenylation Effects 0.000 claims description 5
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 4
- 239000013598 vector Substances 0.000 claims description 4
- 241001465754 Metazoa Species 0.000 claims description 3
- 241000248518 Stylonychia lemnae Species 0.000 claims description 3
- 241000248395 Colpidium Species 0.000 claims description 2
- 241000248332 Colpoda Species 0.000 claims description 2
- 241000248488 Euplotes Species 0.000 claims description 2
- 208000010412 Glaucoma Diseases 0.000 claims description 2
- 241001465965 Holotrichia Species 0.000 claims description 2
- 241000223785 Paramecium Species 0.000 claims description 2
- 241000243198 Peritrichia Species 0.000 claims description 2
- 241000018149 Platyophrya Species 0.000 claims description 2
- 241001491893 Pseudocohnilembus Species 0.000 claims description 2
- 241001467592 Spirotrichea Species 0.000 claims description 2
- 241000248520 Stylonychia Species 0.000 claims description 2
- 241001531212 Suctoria Species 0.000 claims description 2
- 241000223892 Tetrahymena Species 0.000 claims description 2
- 241000868220 Vorticella Species 0.000 claims description 2
- 230000001580 bacterial effect Effects 0.000 claims description 2
- 230000002538 fungal effect Effects 0.000 claims description 2
- 238000000338 in vitro Methods 0.000 claims description 2
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 230000010076 replication Effects 0.000 claims description 2
- 235000013311 vegetables Nutrition 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 description 38
- 241000206602 Eukaryota Species 0.000 description 8
- 210000002231 macronucleus Anatomy 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 108010060309 Glucuronidase Proteins 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 230000021615 conjugation Effects 0.000 description 3
- 210000004940 nucleus Anatomy 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000053187 Glucuronidase Human genes 0.000 description 2
- 108091092195 Intron Proteins 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000013011 mating Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000000392 somatic effect Effects 0.000 description 2
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000000844 transformation Methods 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 2
- DHJFFLKPAYHPHU-BYNIDDHOSA-N 5-bromo-4-chloro-3-indolyl beta-D-glucuronide Chemical compound O1[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 DHJFFLKPAYHPHU-BYNIDDHOSA-N 0.000 description 1
- 102000053642 Catalytic RNA Human genes 0.000 description 1
- 108090000994 Catalytic RNA Proteins 0.000 description 1
- 241000508314 Chlorogonium elongatum Species 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 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 1
- 241000257465 Echinoidea Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000248486 Hypotrichia Species 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 239000007984 Tris EDTA buffer Substances 0.000 description 1
- 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 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000004292 cytoskeleton Anatomy 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000028023 exocytosis Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 230000021121 meiosis Effects 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000008521 reorganization Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 108091092562 ribozyme Proteins 0.000 description 1
- 230000014639 sexual reproduction Effects 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 229940063673 spermidine Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 108091035539 telomere Proteins 0.000 description 1
- 102000055501 telomere Human genes 0.000 description 1
- 210000003411 telomere Anatomy 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- C12N15/63—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
-
- 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
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/89—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microinjection
- C12N15/895—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microinjection using biolistic methods
Definitions
- the present invention relates to a method for expressing a heterologous DNA in a new expression system or host.
- Ciliates are unicellular, animal eukaryotes. They have almost all the typical properties of eukaryotic cells and at the same time offer the advantage that they can be cultured in a similar way to prokaryotes. This means that genetically identical clones can be grown from individual cells by vegetative propagation. In some species, very high cell densities can be achieved in continuous or batch culture.
- ciliates like most eukaryotes, can reproduce sexually. Sexual reproduction, called conjugation in the ciliates, can be induced by bringing together cells of different mating types (mating types are "multiple sexes") under suitable conditions. This property offers the advantage that one can cross ciliates like higher eukaryotes and thereby e.g. Can generate strains that are homozygous for selected traits.
- ciliates In addition to the characteristics typical of most eukaryotes, ciliates have a number of structural and functional features that make them particularly suitable for basic cell biological research as well as for biotechnological applications. Almost all ciliate cells contain two different types of cell nuclei: small, transcription-inactive, mostly diploid micronuclei and mostly very large, DNA-rich macronuclei.
- the micronuclei mainly have generative functions, which means that in the course of conjugation, haploid gamete nuclei develop from them through meiosis.
- the gamete nuclei of two conjugation partners can fuse to form zygote nuclei and create a new cell generation with genetically recombined micronucleus genomes.
- the macronuclei control all somatic processes of the cells. Your genome is transcribed permanently. In the course of macronucleus development, drastic elimination and reorganization processes take place in the genome. In some species, up to 98% of the micronucleus genome is eliminated, intervening sequences are cut out of genes and coding regions can be rearranged completely ("gane scrambling"). In all ciliates examined there, the genes in the macronucleus are more or less strongly amplified. The number of copies can be up to several million, depending on the gene under consideration and the type of ciliate.
- telomeres the end structures of linear DNA molecules and the telomerase ⁇ synthesizing them was first elucidated in ciliates (Blackbum, E.H. (1991): Nature 350, 569-573). It was later shown that these basic processes and structures, which were initially discovered in ciliates due to the special genome structure, are also characteristic of almost all other eukaryotes, are only much more difficult to discover and investigate there.
- Ciliates are unicellular eukaryotes that can be grown in clonal cultures like prokaryotic microorganisms in high cell density with relatively short generation times. 2. Your cells have almost all eukaryotic properties, the prokaryotes are missing, for example in the area of DNA replication, transcription and processing, translation, cytoskeleton and membrane structures, endo- and exocytosis processes, etc.
- ciliates are highly developed eukaryotes that branch off late from the common family tree, their enzymes, structural and membrane proteins and their metabolic pathways are much more similar to the corresponding structures and processes in multicellular eukaryotes (eg humans) than in comparable structures and processes Prokaryotes (bacteria) is the case if there are homologous elements at all.
- heterologous genes or modified genes specific to the species has so far mainly failed because the usual methods available for the transformation of eukaryotes led to sufficiently high transformation rates in ciliates only in a few exceptional cases (Gaertig, J., M. Goravsky (1992 ): Proc. Natl. Acad. Sci. USA 89, 9196-9200).
- there have so far been hardly any suitable selection markers available for attempting transformations with ciliates since some marker systems which are generally used successfully for eukaryotic transformations cannot be used in ciliates (Wünning, IU, Lipps, HJ (1983): EMBO J.2, 1753-1757; Meyers, G., E. Helftenbein (1988 ): Gene 63, 31-40).
- microcarrier bombardment method can be used to transform plant cells surrounded by a rigid and rigid cell wall very effectively (Boynton, JE, NW Gillham, EH Harris, JP Hosler, AM Johnson, AR Jones, BL Randolp-Anderson, D. Robertson, TM Klein, KB Shark, JC Sanford (1988): Science 240, 1534-1537; Klein, TM, L. Kornstein, JC Sanford, ME Fromm (1989): Plant Physiol. 91, 440 -444; Klein, TM, ED Wolf, R. Wu, JC Sanford (1987): Nature 327, 70-73).
- the object of the present invention is to provide a method for expressing a heterologous DNA in a new expression system or host.
- the object is achieved by independent claims 1 and 17 and in particular by the preferred embodiments of subclaims 2 to 16
- the present invention relates to a method for expressing a heterologous DNA in an expression system, characterized in that transformed ciliate cells are used as the expression system
- heterologous DNA sequence is a gene
- ori oil of relication
- ciliate cells are selected from the group Holotrichia, Peritrichia, Spirotrichia and Suctoria.
- ciliate cells are selected from the group Tetrahymena, Paramecium, Colpidium, Colpoda, Glaucoma, Platyophrya, Vorticella, Potomacus, Pseudocohnilembus, Euplotes, Engelmaniella and Stylonychia
- ciliate according to claim 17 characterized in that the transformation was carried out by means of the method of microcarrier bombardment with DNA-loaded gold particles
- a promoter which is active in ciliates and causes the transcription of the heterologous DNA to be expanded
- a termination signal that ends the transcription and is active in ciliates
- a suitable ori origin of the
- step (b) transformation of the ciliate cells with the expression vector according to step (a).
- the expression vector additionally contains a signal sequence which leads to the removal of the gene product from the cell.
- the expression vector additionally contains a signal sequence which leads to the removal of the gene product from the cell.
- Gold particles (1.6 ⁇ m diameter) are suspended at 40 mg / ml in water. 25 ⁇ l of the particle suspension are mixed with 5 ⁇ l DNA solution (conc. 1 ⁇ g / ⁇ l in TE buffer), 25 ⁇ l 2.5 M CaCl 2 solution, 20 ⁇ l 0.1 M spermidine solution under previous tax. After incubation at room temperature for 10 min, the particles are sedimented by centrifugation in the minifuge (12000 rpm). 50 ⁇ l of the supernatant are removed and discarded, the rest is resuspended. 3 ⁇ l of this are pipetted onto the membrane (rupture disk) for the bombardment.
- the plasmid pRT103gus was used for the transformation. It carries ampicillin resistance, the 35S promoter of the Cauliflower Mosaic virus, the coding region of the ß-glucuronidase gene from E. coli and a polyadenylation signal and is successfully used to transform plants.
- BIO-RAD BIOLISTIC Particle Delivery System
- the substrate for the glucuronidase 25 mg of 5-bromo-4-chloro-3-indolyl-glucuronide are dissolved in 4 ml of DMSO and 40 ml of 10 mM EDTA, 100 mM sodium phosphate buffer pH 7.0, 0.1% Triton.
- the transformed ciliate cells were collected by filtration on a filter two days after the transformation experiment.
- the filters with the ciliate cells were incubated in the substrate solution at 32 ° C.
- the cells are partially lysed by the Triton-containing solution.
- Cells that express the transformed glucuronidase gene can be recognized by a clear blue color after a short time. Non-transformed, but otherwise treated control cells show no blue color even after incubation for several hours.
Landscapes
- Genetics & Genomics (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19626564A DE19626564A1 (de) | 1996-07-03 | 1996-07-03 | Genetische Transformation von Ciliatenzellen durch Microcarrier-Bombardement mit DNA beladenen Goldpartikeln |
| DE19626564 | 1996-07-03 | ||
| PCT/EP1997/003472 WO1998001572A1 (de) | 1996-07-03 | 1997-07-02 | Genetische transformation von ciliatenzellen durch microcarrier- bombardement mit dna-beladenen goldpartikeln |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| EP0847444A1 true EP0847444A1 (de) | 1998-06-17 |
Family
ID=7798683
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP97930486A Withdrawn EP0847444A1 (de) | 1996-07-03 | 1997-07-02 | Genetische transformation von ciliatenzellen durch microcarrier- bombardement mit dna-beladenen goldpartikeln |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US6087124A (ja) |
| EP (1) | EP0847444A1 (ja) |
| JP (1) | JP2001510327A (ja) |
| DE (1) | DE19626564A1 (ja) |
| WO (1) | WO1998001572A1 (ja) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9127285B2 (en) | 2012-02-22 | 2015-09-08 | The University Of Chicago | Genetically altered ciliates and uses thereof |
Families Citing this family (58)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000023604A1 (de) * | 1998-10-21 | 2000-04-27 | Celanese Ventures Gmbh | Expressionsvektoren enthaltend regulative sequenzen aus stylonychia lemnae zur heterologen proteinexpression in eukaryontischen protisten und ein verfahren zur identifizierung solcher regulativen sequenzen |
| US7026156B1 (en) | 1999-02-04 | 2006-04-11 | The University Of Georgia Research Foundation, Inc. | Diagnostic and protective antigen gene sequences of ichthyophthirius |
| US7326568B2 (en) * | 1999-02-04 | 2008-02-05 | University Of Georgia Research Foundation, Inc. | Recombinant expression of heterologous nucleic acids in protozoa |
| US6846481B1 (en) | 1999-02-04 | 2005-01-25 | University Of Georgia Research Foundation, Inc. | Recombinant expression of heterologous nucleic acids in protozoa |
| IL150414A0 (en) | 2000-02-09 | 2002-12-01 | Basf Ag | Novel elongase gene and method for producing multiple-unsaturated fatty acid |
| EP1728870A3 (en) | 2000-04-07 | 2007-01-10 | BASF Plant Science GmbH | Transcription factor stress-related proteins and methods of use in plants |
| KR20020028419A (ko) * | 2000-10-10 | 2002-04-17 | 서만석 | 유전자 또는 유전자 백신의 전달방법 |
| EP1395108B1 (en) | 2001-03-16 | 2012-01-11 | BASF Plant Science GmbH | Sugar and lipid metabolism regulators in plants |
| AU2002305858B2 (en) | 2001-06-04 | 2007-08-23 | Basf Plant Science Gmbh | Sugar and lipid metabolism regulators in plants II |
| DE60231717D1 (de) | 2001-08-10 | 2009-05-07 | Basf Plant Science Gmbh | Zucker- und lipidmetabolismusregulatoren bei pflanzen iii |
| CA2459961A1 (en) | 2001-09-05 | 2003-03-13 | Basf Plant Science Gmbh | Protein phosphatase stress-related polypeptides and methods of use in plants |
| ES2365912T3 (es) | 2001-11-09 | 2011-10-13 | Basf Plant Science Gmbh | Polipéptidos del factor de transcripción relacionados con el estrés y métodos de uso en plantas. |
| DE10212892A1 (de) | 2002-03-20 | 2003-10-09 | Basf Plant Science Gmbh | Konstrukte und Verfahren zur Regulation der Genexpression |
| DE10214413A1 (de) * | 2002-03-30 | 2003-10-09 | Nutrinova Gmbh | Expression von rekombinanten humanen Proteinen in Tetrahymena |
| WO2004013304A2 (en) | 2002-08-02 | 2004-02-12 | Basf Plant Science Gmbh | Sugar and lipid metabolism regulators in plants iv |
| AU2003294924A1 (en) | 2002-12-20 | 2004-07-14 | Metanomics Gmbh & Co. Kgaa | Method for producing aminoacids |
| BRPI0407514A (pt) | 2003-02-17 | 2006-02-14 | Metanomics Gmbh | método para preparar um organismo não humano, polinucleotìdeo, método para preparar um vetor, vetor, célula hospedeira, polipeptìdeo, anticorpo, ácido polinucléico de anti-sentido, método para preparar uma planta, célula vegetal, tecido vegetal transgênicos, célula de um animal útil, animal útil ou um microorganismo transgênico organismo não humano, microorganismo transgênico, rendimento ou um material de propagação de uma planta ou de um animal útil, biomassa de um microorganismo, método para preparar produtos quìmicos finos, e, uso de polipeptìdeo |
| BRPI0407138A (pt) | 2003-02-27 | 2006-01-10 | Basf Plant Science Gmbh | Sequência de ácido nucleico isolada, sequência de aminoácido, construção de gene, vetor, organismo transgênico não humano, processo para produzir ácidos graxos poliinsaturados, óleo, lipìdeo, ou um ácido graxo poliinsaturado ou uma fração dos mesmos, composições de óleo, de lipìdeos, ou de ácido graxo, e, uso do óleo, lipìdeos ou ácidos graxos ou de composições de óleo, de lipìdeos ou de ácido graxo |
| ES2421138T3 (es) | 2003-03-31 | 2013-08-29 | University Of Bristol | Nuevas aciltransferasas vegetales específicas para ácidos grasos poliinsaturados de cadena larga |
| CA2521754A1 (en) | 2003-04-15 | 2004-10-28 | Basf Plant Science Gmbh | Nucleic acid sequences encoding proteins associated with abiotic stress response and plant cells and plants with increased tolerance to environmental stress |
| WO2005014828A2 (en) | 2003-08-01 | 2005-02-17 | Basf Plant Science Gmbh | Process for the production of fine chemicals in plants |
| EP2166069B2 (de) | 2003-08-01 | 2024-01-24 | BASF Plant Science GmbH | Verfahren zur Herstellung mehrfach ungesättigter Fettsäuren in transgenen Organismen |
| CN1898389A (zh) | 2003-12-23 | 2007-01-17 | 巴斯福植物科学有限公司 | 植物中的糖及脂质代谢调节剂vi |
| WO2005083053A2 (de) | 2004-02-27 | 2005-09-09 | Basf Plant Science Gmbh | Verfahren zur herstellung von ungesättigten omega-3-fettsäuren in transgenen organismen |
| JP4567047B2 (ja) | 2004-02-27 | 2010-10-20 | ビーエーエスエフ プラント サイエンス ゲーエムベーハー | トランスジェニック植物における多価不飽和脂肪酸の製造方法 |
| AU2005262493A1 (en) | 2004-06-16 | 2006-01-19 | Basf Plant Science Gmbh | Nucleic acid molecules encoding wrinkled1-like polypeptides and methods of use in plants |
| EP2080769A3 (en) | 2004-07-02 | 2010-12-01 | Metanomics GmbH | Process for the production of fine chemicals |
| BRPI0514003A (pt) | 2004-07-31 | 2008-05-20 | Metanomics Gmbh | métodos para preparar um organismo não humano, para preparar um vetor, uma planta transgênica, célula vegetal, tecido vegetal, célula de um animal útil, animal útil ou um microorganismo transgênico, e produtos quìmicos finos, e para identificar um produto de gene, polinucleotìdeo, vetor, célula hospedeira, anticorpo, ácido nucleico antisentido, organismo não humano, microorganismo transgênico, semente, tubérculo ou material de propagação de uma planta, biomassa de um microorganismo, célula vegetal, organela de célula vegetal, tecido vegetal, planta ou parte destes, e, usos de um polipeptìdeo, e da molécula de ácido nucleico |
| EP1781784A2 (en) | 2004-08-02 | 2007-05-09 | BASF Plant Science GmbH | Method for isolation of transcription termination sequences |
| EP2339007A1 (en) | 2004-09-20 | 2011-06-29 | BASF Plant Science GmbH | Arabidopsis genes encoding proteins involved in sugar and lipid metabolism and methods of use |
| US8324455B2 (en) | 2004-09-24 | 2012-12-04 | Basf Plant Science Gmbh | Nucleic acid sequences encoding proteins associated with abiotic stress response and plant cells and plants with increased tolerance to environmental stress |
| EP2096177A3 (en) | 2004-12-17 | 2010-01-13 | Metanomics GmbH | Process for the production of lutein |
| CA2598792A1 (en) * | 2005-03-02 | 2006-09-08 | Metanomics Gmbh | Process for the production of fine chemicals |
| US20140199313A1 (en) | 2005-03-02 | 2014-07-17 | Metanomics Gmbh | Process for the Production of Fine Chemicals |
| DE102005013779A1 (de) | 2005-03-22 | 2006-09-28 | Basf Plant Science Gmbh | Verfahren zur Herstellung von mehrfach ungesättigten C20- und C22-Fettsäuren mit mindestens vier Doppelbindungen in transgenen Pflanzen |
| US7880055B2 (en) | 2005-06-17 | 2011-02-01 | Basf Plant Science Gmbh | Lecitin-like protein kinase stress-related polypeptides and methods of use in plants |
| US8097769B2 (en) | 2005-07-18 | 2012-01-17 | Basf Plant Science Gmbh | Yield increase in plants overexpressing the ACCDP genes |
| EP2333087A1 (en) | 2005-07-18 | 2011-06-15 | BASF Plant Science GmbH | Yield increase in plants overexpressing the SHSRP genes |
| DE102005038036A1 (de) | 2005-08-09 | 2007-02-15 | Basf Plant Science Gmbh | Verfahren zur Herstellung von Arachidonsäure und/oder Eicosapentaensäure in transgenen Nutzpflanzen |
| MX2008001513A (es) | 2005-08-12 | 2008-04-04 | Basf Plant Science Gmbh | Secuencias de acidos nucleicos que codifican proteinas asociadas con respuesta a estres abiotico y celulas vegetales y plantas con aumento de tolerancia a estres ambiental. |
| US7723574B2 (en) | 2005-11-24 | 2010-05-25 | Basf Plant Science Gmbh | Process for the production of Δ5-unsaturated fatty acids in transgenic organisms |
| EP2522722A3 (en) | 2005-12-09 | 2012-12-12 | BASF Plant Science GmbH | Nucleic acid molecules encoding polypeptides involved in regulation of sugar and lipid metabolism and methods of use VIII |
| GB0603160D0 (en) | 2006-02-16 | 2006-03-29 | Rothamsted Res Ltd | Nucleic acid |
| EP2010661A2 (en) | 2006-03-24 | 2009-01-07 | BASF Plant Science GmbH | Proteins associated with abiotic stress response and homologs |
| EP2090662A3 (en) | 2006-04-05 | 2012-10-31 | Metanomics GmbH | Process for the production of a fine chemical |
| EP2497778A3 (en) | 2006-05-31 | 2012-12-12 | Metanomics GmbH | Manipulation of the nitrogen metabolism |
| ES2531391T3 (es) | 2006-10-06 | 2015-03-13 | Basf Plant Science Gmbh | Delta-5 desaturasas y procedimiento para la preparación de ácidos grasos poliinsaturados en organismos transgénicos no humanos |
| US8426676B2 (en) | 2007-05-04 | 2013-04-23 | Basf Plant Science Gmbh | Seed enhancement by combinations of pyruvate kinases |
| DE112008001453T5 (de) | 2007-05-22 | 2010-04-29 | Basf Plant Science Gmbh | Pflanzenzellen und Pflanzen mit erhöhter Toleranz und/oder Resistenz gegenüber Umweltstress und erhöhter Biomasseproduktion-KO |
| US20110041215A1 (en) * | 2007-12-17 | 2011-02-17 | Basf Plant Science Gmbh | Lipid Metabolism Protein And Uses Thereof I (BZIP Transcription Factor) |
| BRPI0821231A2 (pt) | 2007-12-17 | 2014-12-23 | Basf Plant Science Gmbh | Polinucleotídeo, vetor , célula hospedeira, método para a fabricação de um polipeptídeo, polipeptídeo, anticorpo, organismo não humano transgênico, e, métodos para a fabricação de um lipídeo ou um ácido graxo e para a fabricação de uma planta. |
| AR069894A1 (es) * | 2007-12-21 | 2010-02-24 | Basf Plant Science Gmbh | Plantas con mayor rendimiento produciendo un knock out (silenciamiento) en genes relacionados con la eficiencia en el uso de nitrogeno (ko nue)" |
| AU2009326129B2 (en) | 2008-12-12 | 2016-05-12 | Basf Plant Science Gmbh | Desaturases and process for the production of polyunsaturated fatty acids in transgenic organisms |
| EP2408911B1 (en) | 2009-03-20 | 2017-01-25 | Tetragenetics, Inc. | Polypeptide expression in ciliates |
| GB201003701D0 (en) | 2010-03-05 | 2010-04-21 | Cilian Ag | System for the expression of a protein |
| WO2013024121A2 (en) | 2011-08-18 | 2013-02-21 | Basf Plant Science Company Gmbh | Increase of sucrose transporter activity in the seeds of plants |
| AU2014369232B2 (en) | 2013-12-17 | 2021-02-18 | Basf Plant Science Company Gmbh | Methods for conversion of the substrate specificity of desaturases |
| CN106070081A (zh) * | 2016-07-08 | 2016-11-09 | 扬州大学 | 一种用于室内人工饲养襀翅目昆虫的方法 |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5036006A (en) * | 1984-11-13 | 1991-07-30 | Cornell Research Foundation, Inc. | Method for transporting substances into living cells and tissues and apparatus therefor |
| US5179022A (en) * | 1988-02-29 | 1993-01-12 | E. I. Du Pont De Nemours & Co. | Biolistic apparatus for delivering substances into cells and tissues in a non-lethal manner |
| DE4238842A1 (de) * | 1992-11-17 | 1994-05-19 | Arno Prof Dr Tiedtke | Hochzelldichte Fermentation von Ciliaten zur Gewinnung von Naturstoffen |
| ATE278772T1 (de) * | 1994-10-21 | 2004-10-15 | Nutrinova Gmbh | Verfahren zur kultivierung lipidabhängiger tetrahymeniden und ein verfahren zur herstellung von biogenen wertstoffen aus lipidabhängigen tetrahymeniden |
-
1996
- 1996-07-03 DE DE19626564A patent/DE19626564A1/de not_active Withdrawn
-
1997
- 1997-07-02 US US09/029,444 patent/US6087124A/en not_active Expired - Lifetime
- 1997-07-02 WO PCT/EP1997/003472 patent/WO1998001572A1/de not_active Application Discontinuation
- 1997-07-02 EP EP97930486A patent/EP0847444A1/de not_active Withdrawn
- 1997-07-02 JP JP50474898A patent/JP2001510327A/ja active Pending
Non-Patent Citations (1)
| Title |
|---|
| See references of WO9801572A1 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9127285B2 (en) | 2012-02-22 | 2015-09-08 | The University Of Chicago | Genetically altered ciliates and uses thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2001510327A (ja) | 2001-07-31 |
| US6087124A (en) | 2000-07-11 |
| DE19626564A1 (de) | 1998-01-08 |
| WO1998001572A1 (de) | 1998-01-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO1998001572A1 (de) | Genetische transformation von ciliatenzellen durch microcarrier- bombardement mit dna-beladenen goldpartikeln | |
| DE3689802T2 (de) | "Hervorrufung somatischer Veränderungen in Pflanzen durch Verwendung von minussträngigen RNAs". | |
| Macgregor et al. | The biological significance of variation in satellite DNA and heterochromatin in newts of the genus Triturus: an evolutionary perspective | |
| DE60222857T2 (de) | Concatemere unterschiedlich exprimierter multipler gene | |
| Bonnefoy et al. | Genetic transformation of Saccharomyces cerevisiae mitochondria | |
| DE69233380T2 (de) | Transgene pflanzen mit geändertem polyolgehalt | |
| WO1987006614A1 (en) | Electric field mediated dna transformation of plant cells and organelles | |
| Zaid et al. | Glossary of biotechnology and genetic engineering | |
| CN102498220B (zh) | 甘蔗的着丝粒序列和微型染色体 | |
| CN109456994B (zh) | 一种厚壳贻贝眼点幼虫的电穿孔转染方法 | |
| Heenatigala et al. | Development of efficient protocols for stable and transient gene transformation for Wolffia globosa using Agrobacterium | |
| JPH05508547A (ja) | ライブラリースクリーニング法 | |
| DE3786771T2 (de) | Für inaktivierte IMP-Dehydrogenase kodierende DNS. | |
| DE4309203C1 (de) | Verfahren zur Produktion von transgenischen Pflanzen | |
| Berger et al. | An apparent lack of non-transcribed spacers in rDNA of a green alga | |
| CN102203238B (zh) | 鲨肌醇产生细胞和使用该细胞的鲨肌醇制造方法 | |
| Berger et al. | 80 S ribosomes in Acetabularia major redundancy of rRNA cistrons | |
| CN102190715B (zh) | 一种植物砷抗性相关的蛋白及编码基因及其应用 | |
| DE19943383A1 (de) | Regulatorische Sequenzen und Expressionskassetten für Hefen | |
| JPS59501195A (ja) | 原核生物における遺伝子工学 | |
| Guilfoyle et al. | Synthesis of 5S rRNA and putative precursor tRNAs in nuclei isolated from wheat embryos | |
| Bilinski et al. | Apomixis in Saccharomyces cerevisiae and other eukaryotic micro-organisms | |
| Drew | Induction of resistance to A-methopterin in Diplococcus pneumoniae by deoxyribonucleic acid | |
| EP1357190B1 (de) | Verfahren und Marker zur einfachen Transformation und Selektion von rekombinanten Ciliaten | |
| DE10024334A1 (de) | Verfahren zum Einbringen von Nukleinsäuren in eine Zelle (Transfektion) mittels Calciumphosphat sowie transfizierte Zelle |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
| AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LI LU NL PT SE |
|
| 17P | Request for examination filed |
Effective date: 19980715 |
|
| RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: AXIVA GMBH |
|
| RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: CELANESE VENTURES GMBH |
|
| 17Q | First examination report despatched |
Effective date: 20011218 |
|
| RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: NUTRINOVA NUTRITION SPECIALTIES & FOOD INGREDIENTS |
|
| STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
| 18D | Application deemed to be withdrawn |
Effective date: 20021202 |