WO2001007624A2 - Dna sequences which are suited for specifically detecting pseudomonas putida kt2440 - Google Patents
Dna sequences which are suited for specifically detecting pseudomonas putida kt2440 Download PDFInfo
- Publication number
- WO2001007624A2 WO2001007624A2 PCT/EP2000/007238 EP0007238W WO0107624A2 WO 2001007624 A2 WO2001007624 A2 WO 2001007624A2 EP 0007238 W EP0007238 W EP 0007238W WO 0107624 A2 WO0107624 A2 WO 0107624A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- seq
- dna sequences
- pseudomonas putida
- suited
- specifically detecting
- 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.)
- Ceased
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/21—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Pseudomonadaceae (F)
-
- 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
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
Definitions
- the invention relates to DN ⁇ sequences which are suitable for the specific detection of Pseudomonas putida KT2440.
- the Gram-negative soil bacterium Pseudomonas putida strain T2440 has been tested by the United States Department of Health (NIH [rec]) and has been declared a safety strain. As concerns about both closed and open biotechnological use of bacterial strains related to plant, animal or human palhogenic bacteria increase, the importance of this safety strain for various applications increases.
- the object of the invention is therefore to provide DNA sequences which are suitable for the specific detection of Pseudomonas putida KT2440, in particular in the presence of other bacteria of the genus Pseudomonas, for example Pseudomonas aeruginosa.
- Pseudomonas putida KT2440 is characterized by a very high catabolic diversity and by a high survivability and adaptability to different locations (rhizosphere and soil).
- this bacterium can be used for working with recombined DNA in a wide variety of areas of biotechnology (e.g. as a microbial production strain, for biological remediation processes etc.) and biomedicine (e.g. as a microbial vaccine carrier).
- biotechnology e.g. as a microbial production strain, for biological remediation processes etc.
- biomedicine e.g. as a microbial vaccine carrier.
- the use of the genes or nucleic acid sequences or modified modifications thereof disclosed according to the invention and the expression products derived therefrom for detection are the basis for the genetic safety of this strain.
- DNA sequences can be broken down into known and optimized expression vectors using customary molecular biological methods (cf., for example, BJ Sambrook, EF Fritsch, T. Maniatis, "Molecular Cloning, A Laboratory Manual", Cold Spring Harbor Laboratory Press, New York, 1989) insert, which enables the appropriate transformation, selection and cloning of cells which are then able to synthesize the expression products by fermentation. If an overproducing cell clone is chosen, the desired expression products can easily be produced and obtained in large quantities.
- the expression of the coding sequences can be modulated, in particular in bacteria of the genus Pseudomonas, for example the Pseudomonas putida or Pseudomonas fluorescence group. Modulation is generally used to describe understood the influence of expression, e.g. B. switching on and off or increasing / decreasing expression.
- the invention thus further relates to a recombined expression vector according to claim 2, thus transformed cells according to claim 3, a method for producing expression products according to claim 5, expression products or partial expression products according to claim 6, synthetic peptides or proteins according to claim 7, poly or monoclonal antibodies according to claims 8 and 9, hybridoma cells according to claim 10, transgenic plants according to claim 13 and the use of the DNA sequences as probes or PCR primers according to claim 15 and a method for the specific detection of Pseudomonas putida KT2440 according to claim 16 ,
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to DNA sequences which are suited for specifically detecting Pseudomonas putida KT2440.
Description
DNA-Sequenzen, die sich zum spezifischen Nachweis von Pseudomonas putida KT2440 eignen . DNA sequences that are suitable for the specific detection of Pseudomonas putida KT2440.
Die Erfindung betrifft DNΛ-Sequenzen, die sich zum spezifischen Nachweis von Pseudomonas putida KT2440 eignen.The invention relates to DNΛ sequences which are suitable for the specific detection of Pseudomonas putida KT2440.
Einleitung und Stand der TechnikIntroduction and state of the art
Das Gram-negative Bodenbakterium Pseudomonas putida Stamm T2440 ist durch die US-amerikanische Gesundheitsbehörde (NIH [rec]) geprüft und als Sicherheitsstamm deklariert worden. Mit der Zunahme der Bedenken im Hinblick auf sowohl die geschlossene als auch die offene biotechnologische Verwendung von Bakterienstämmen, die mit pflanzen-, tier- oder humanpalhogenen Bakterien verwandt sind, erhöht sich die Bedeutung dieses Sicherheitsstammes für verschiedene Anwendungen.The Gram-negative soil bacterium Pseudomonas putida strain T2440 has been tested by the United States Department of Health (NIH [rec]) and has been declared a safety strain. As concerns about both closed and open biotechnological use of bacterial strains related to plant, animal or human palhogenic bacteria increase, the importance of this safety strain for various applications increases.
Ein wesentlicher Sicherheitsaspekl von Pseudomonas putida KT2440 ist das Fehlen von potentiell pathogen Eigenschaften. Zur Zeit ist die Verwendung von drei Bakteriengattungen, nämlich Escherichia coli (Stämme chi-1776, MRC und Kl 2), der asporogenen, thyminabhängigen Mutante des Bacillus-subtilis-Sta mes 168 und des oben beschriebenen Pseudomonαs-putidα- KT2440-Stammes, jeweils mit den entsprechenden genetischen Vektoren , als "biologische Sicherheitsmaßnahme" anerkannt. Die Verwendung dieser Bakterien für gentechnische Arbeiten gemäß der GenTSV reduziert in beträchtlichem Maße das Gefährdungspotential und beeinflußt somit positiv die behördliche Gesamtbewertung der gentechnischen Arbeiten in gentechnischen Anlagen.An essential safety aspect of Pseudomonas putida KT2440 is the lack of potentially pathogenic properties. Currently, the use of three bacterial genera, namely Escherichia coli (strains chi-1776, MRC and Kl 2), the asporogenic, thymine-dependent mutant of the Bacillus subtilis strain 168 and the Pseudomonαs putidα KT2440 strain described above, is in each case with the appropriate genetic vectors, recognized as a "biological safety measure". The use of these bacteria for genetic engineering work in accordance with the GenTSV considerably reduces the risk potential and thus has a positive influence on the overall official assessment of genetic engineering work in genetic engineering plants.
Beschreibung der ErfindungDescription of the invention
Erfindungsgemäß wurde gefunden, daß es zahlreiche Gene gibt, die ausschließlich in diesem Organismus vorhanden sind und keine signifikanten Homologien zu Genen aus bereits bekannten Organismen aufweisen. Überraschenderweise wurde gefunden, daß diese Gene nicht in dem mit Pseudomonas putida KT2440 nahe verwandten, aber humanpa- thogen Bakterienstamm Pseudomonas aeruginosa vorhanden sind. Diese für Pseudomonas putida KT2440 charakteristischen Gene und die davon abzuleitenden Funktionen sind für die Sicherheit des Bakteriums maßgeblich.According to the invention, it was found that there are numerous genes which are only present in this organism and have no significant homologies to genes from organisms which are already known. Surprisingly, it was found that these genes are not present in the bacterial strain Pseudomonas aeruginosa, which is closely related to Pseudomonas putida KT2440, but is pathogenic to humans. These genes, which are characteristic of Pseudomonas putida KT2440, and the functions to be derived from them are essential for the safety of the bacterium.
Aufgabe der Erfindung ist somit die Bereitstellung von DNA-Sequenzen gemäß Patentanspruch 1, die sich zum spezifischen Nachweis von Pseudomonas putida KT2440 eignen, und zwar insbesondere in Gegenwart von anderen Bakterien der Gattung Pseudomonas, beispielsweise Pseudomonas aeruginosa.The object of the invention is therefore to provide DNA sequences which are suitable for the specific detection of Pseudomonas putida KT2440, in particular in the presence of other bacteria of the genus Pseudomonas, for example Pseudomonas aeruginosa.
Im folgenden bedeutet die Angabe "ORF" einen offenen Leserahmen (open reading fra- me).In the following, the indication "ORF" means an open reading frame.
Durch die Bereitstellung derartiger DNA-Sequenzen lassen sich folgende Vorteile erzielen. Im Gegensatz zu den beiden anderen biologischen Sicherheitsstämmen zeichnet sich Pseudomonas putida KT2440 durch eine sehr hohe katabolische Vielfalt und durch eine hohe Überlebens- und Anpassungsfähigkeit an verschiedene Standorte (Rhizosphäre und Boden) aus. In Kombination mit den anerkannten Vektorsystemen kann dieses Bakterium für Arbeiten mit rekombinierter DNS in den verschiedensten Bereichen der Biotechnologie (z. B. als mikrobieller Produktionsstamm, für biologische Sanierungsverfahren etc.) und der Biomedizin (z. B. als mikrobieller Impfstoffträger) eingesetzt werden. Die Verwendung der erfindungsgemäß offenbarten Gene bzw. Nucleinsäuresequenzen oder modifizierten Abwandlungen davon und der davon abgeleiteten Expressionsprodukte zum Nachweis sind die Basis für die genetische Sicherheit dieses Stammes.The following advantages can be achieved by providing such DNA sequences. In contrast to the other two biological security strains, Pseudomonas putida KT2440 is characterized by a very high catabolic diversity and by a high survivability and adaptability to different locations (rhizosphere and soil). In combination with the recognized vector systems, this bacterium can be used for working with recombined DNA in a wide variety of areas of biotechnology (e.g. as a microbial production strain, for biological remediation processes etc.) and biomedicine (e.g. as a microbial vaccine carrier). The use of the genes or nucleic acid sequences or modified modifications thereof disclosed according to the invention and the expression products derived therefrom for detection are the basis for the genetic safety of this strain.
Die DNA-Sequenzen lassen sich mit üblichen molekularbiologischen Methoden (vgl. z. B. J. Sambrook, E. F. Fritsch, T. Maniatis, "Molecular Cloning, A Laboratory Manual", Cold Spring Harbour Laboratory Press, New York, 1989) in bekannte und optimierte Expressionsvektoren insertieren, wodurch die entsprechende Transformation, Selektion und Klonierung von Zellen möglich ist, die dann zur Synthese der Expressionsprodukte durch Fermentation in der Lage sind. Wenn ein überproduzierender Zellklon gewählt wird, lassen sich die gewünschten Expressionsprodukte leicht in großen Mengen herstellen und gewinnen.The DNA sequences can be broken down into known and optimized expression vectors using customary molecular biological methods (cf., for example, BJ Sambrook, EF Fritsch, T. Maniatis, "Molecular Cloning, A Laboratory Manual", Cold Spring Harbor Laboratory Press, New York, 1989) insert, which enables the appropriate transformation, selection and cloning of cells which are then able to synthesize the expression products by fermentation. If an overproducing cell clone is chosen, the desired expression products can easily be produced and obtained in large quantities.
Die Kenntnis der DNA-Sequenzen gestattet die gezielte Mutagenese ("site-directed mutagenesis") mit üblichen gentechnischen Methoden und somit die Konstruktion von optimierten Expressionsprodukten ("protein engineering").Knowledge of the DNA sequences permits targeted mutagenesis ("site-directed mutagenesis") using conventional genetic engineering methods and thus the construction of optimized expression products ("protein engineering").
Ferner ist die Modulation der Expression der kodierenden Sequenzen, insbesondere in Bakterien der Gattung Pseudomonas, beispielsweise der Pseudomonas-putida- oder Pseudomonas-fluoreszenz-Gτυppe, möglich. Unter Modulation wird allgemein eine Be- einflußung der Expression verstanden, z. B. das An- und Ausschalten oder die Erhöhung/Erniedrigung der Expression.Furthermore, the expression of the coding sequences can be modulated, in particular in bacteria of the genus Pseudomonas, for example the Pseudomonas putida or Pseudomonas fluorescence group. Modulation is generally used to describe understood the influence of expression, e.g. B. switching on and off or increasing / decreasing expression.
Die Erfindung betrifft somit ferner einen rekombinierten Expressionsvektor nach Patentanspruch 2, damit transformierte Zellen nach Patentanspruch 3, ein Verfahren zur Herstellung von Expressionsprodukten nach Patentanspruch 5, Expressionsprodukte- bzw. Teilexpressionsprodukte nach Patentanspruch 6, synthetische Peptide bzw. Proteine nach Patentanspruch 7, poly- bzw. monoklonale Antikörper nach Patentanspruch 8 bzw. 9, Hybridomzellen nach Patentanspruch 10, transgene Pflanzen nach Patentanspruch 13 sowie die Verwendung der DNA-Sequenzen als Sonden bzw. PCR-Primer nach Patentanspruch 15 und ein Verfahren zum spezifischen Nachweis von Pseudomonas putida KT2440 nach Patentanspruch 16.The invention thus further relates to a recombined expression vector according to claim 2, thus transformed cells according to claim 3, a method for producing expression products according to claim 5, expression products or partial expression products according to claim 6, synthetic peptides or proteins according to claim 7, poly or monoclonal antibodies according to claims 8 and 9, hybridoma cells according to claim 10, transgenic plants according to claim 13 and the use of the DNA sequences as probes or PCR primers according to claim 15 and a method for the specific detection of Pseudomonas putida KT2440 according to claim 16 ,
Vorteilhafte Ausführungsformen der Erfindung sind Gegenstand der Unteransprüche.Advantageous embodiments of the invention are the subject of the dependent claims.
Verfahren zum Anzüchten beliebiger Zellen, zur Isolierung der DNA daraus sowie zu deren Amplifikation, z. B. durch die Polymerasekettenreaktion, und deren Sequenzierung sind im Stand der Technik bekannt und bedürfen keiner weiteren Erläuterung. Das gleiche gilt für die Rekombination von DNA-Sequenzen, die Konstruktion/Rekombination von geeigneten Expressionsvektoren und die Transformation/Transfektion von beliebigen Zellen mit molekularbiologischen Techniken, die Identifikation/Selektion von geeigneten Klonen und deren Anzüchtung und die Gewinnung, Reinigung und Charakterisierung der von diesen Klonen produzierten Expressionsprodukte (vgl. beispielsweise das oben zitierte Standardwerk von Maniatis et al.; D. S. T. Nicholl, "Gentechnische Methoden", Spektrum Akademischer Verlag, 1995; C. R. Newton, A. Graham, "PCR", Spektrum Akademischer Verlag, 1994; F. Lotspeich/H. Zorbas (Hrsg.), "Bioanalytik", Spektrum Akademischer Verlag, 1998). Auch die Herstellung und Markierung von poly- oder monoklonalen Antikörpern bzw. die Herstellung der die letzteren bildenden Hybridome ist seit langem bekannt (vgl. beispielsweise: E. Harlow, D. Lane, "Antibodies, A Labo- ratory Manual", Cold Spring Harbor Laboratory, 1988; E. Lidell, I. Weeks, "Antikörper- Techniken", Spektrum Akademischer Verlag, 1996).Process for culturing any cells, isolating the DNA therefrom and amplifying them, e.g. B. by the polymerase chain reaction, and their sequencing are known in the prior art and need no further explanation. The same applies to the recombination of DNA sequences, the construction / recombination of suitable expression vectors and the transformation / transfection of any cells using molecular biological techniques, the identification / selection of suitable clones and their cultivation and the extraction, purification and characterization of these Cloning produced expression products (see, for example, the standard work by Maniatis et al cited above; DST Nicholl, "Genetic Methods", Spektrum Akademischer Verlag, 1995; CR Newton, A. Graham, "PCR", Spektrum Akademischer Verlag, 1994; F. Lotspeich / H. Zorbas (ed.), "Bioanalytik", Spektrum Akademischer Verlag, 1998). The production and labeling of poly- or monoclonal antibodies and the production of the hybridomas forming the latter have also been known for a long time (cf. for example: E. Harlow, D. Lane, "Antibodies, A Labo- Ratory Manual ", Cold Spring Harbor Laboratory, 1988; E. Lidell, I. Weeks," Antibody Techniques ", Spectrum Academic Publisher, 1996).
Der spezifische Nachweis von Genen oder Nucleinsäuresequenzen oder Proteinen oder von diese enthaltenden oder sezemierenden Zellen zur Identifizierung der Zellen oder diese enthaltenden multizellulären Organismen ist ebenfalls bekannt (vgl. die oben angegebenen Literaturstellen). The specific detection of genes or nucleic acid sequences or proteins or of cells containing or secreting them for the identification of the cells or multicellular organisms containing them is also known (cf. the references cited above).
Claims
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU61603/00A AU6160300A (en) | 1999-07-27 | 2000-07-27 | Dna sequences which are suited for specifically detecting pseudomonas putida kt2440 |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19935088A DE19935088A1 (en) | 1999-07-27 | 1999-07-27 | DNA sequences that are suitable for the specific detection of Pseudomonas putida KT2440 |
| DE19935088.4 | 1999-07-27 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| WO2001007624A2 true WO2001007624A2 (en) | 2001-02-01 |
| WO2001007624A3 WO2001007624A3 (en) | 2001-08-02 |
Family
ID=7916126
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/EP2000/007238 Ceased WO2001007624A2 (en) | 1999-07-27 | 2000-07-27 | Dna sequences which are suited for specifically detecting pseudomonas putida kt2440 |
Country Status (3)
| Country | Link |
|---|---|
| AU (1) | AU6160300A (en) |
| DE (1) | DE19935088A1 (en) |
| WO (1) | WO2001007624A2 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7711849B2 (en) | 2000-04-03 | 2010-05-04 | West Notifications Group, Inc. | Individual XML message processing platform |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2026333A6 (en) * | 1990-10-09 | 1992-04-16 | Consejo Superior Investigacion | System for detection and enumeration of the bacteria Pseudomonas putida KT2440 |
| CA2175435A1 (en) * | 1996-04-30 | 1997-10-31 | Joseph Lam | Proteins involved in the synthesis and assembly of o-antigen in pseudomonas aeruginosa |
| US5750848A (en) * | 1996-08-13 | 1998-05-12 | Monsanto Company | DNA sequence useful for the production of polyhydroxyalkanoates |
-
1999
- 1999-07-27 DE DE19935088A patent/DE19935088A1/en not_active Withdrawn
-
2000
- 2000-07-27 AU AU61603/00A patent/AU6160300A/en not_active Abandoned
- 2000-07-27 WO PCT/EP2000/007238 patent/WO2001007624A2/en not_active Ceased
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7711849B2 (en) | 2000-04-03 | 2010-05-04 | West Notifications Group, Inc. | Individual XML message processing platform |
| US7809855B2 (en) | 2000-04-03 | 2010-10-05 | West Notifications Group, Inc. | Individual XML message processing platform |
| US8386569B2 (en) | 2000-04-03 | 2013-02-26 | West Corporation | Individual XML message processing platform |
| US8706904B1 (en) | 2000-04-03 | 2014-04-22 | West Notifications, Inc. | Individual XML message processing platform |
| US9300608B1 (en) | 2000-04-03 | 2016-03-29 | West Notifications, Inc. | Individual XML message processing platform |
Also Published As
| Publication number | Publication date |
|---|---|
| DE19935088A1 (en) | 2001-02-01 |
| AU6160300A (en) | 2001-02-13 |
| WO2001007624A3 (en) | 2001-08-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| DE69730514T2 (en) | amidase | |
| Marilley et al. | Phylogenetic diversity of bacterial communities differing in degree of proximity of Lolium perenne and Trifolium repens roots | |
| DE3629890A1 (en) | MICROORGANISMS AND PLASMIDES FOR THE 2,4-DICHLORPHENOXYACETIC ACID (2,4-D) MONOOXIGENASE - FORMATION AND METHOD FOR PRODUCING THIS PLASMIDE AND STEM | |
| DE69631787T2 (en) | PROCESS FOR SCREENING ENZYMATIC ACTIVITY | |
| DE4447471A1 (en) | Sequences for proteins with saccharose-isomerase activity | |
| Hahn et al. | Synthetic oligonucleotide probes for identification of Frankia strains | |
| WO1994021788A2 (en) | Testing for infestation of rapeseed and other cruciferae by the fungus leptosphaeria maculans (blackleg infestation) | |
| WO2001007624A2 (en) | Dna sequences which are suited for specifically detecting pseudomonas putida kt2440 | |
| DE69635739T2 (en) | USE OF A SEC DEPENDENT SECRETION SYSTEM FOR THE SECRETION OF PROTEINS, WHICH ARE SUITABLY SECRETED BY A SEC INDEPENDENT SECRETION SYSTEM | |
| DE69835984T2 (en) | Oligonucleotides suitable for the detection of lactic acid bacteria | |
| Springer et al. | A 23S rRNA targeted specific hybridization probe for Bradyrhizobium japonicum | |
| WO2001007621A2 (en) | Dna sequences coding for abc transporters | |
| WO2001007622A2 (en) | DNA SEQUENCES THAT ENCODE TYPE II (xcp) SECRETION SYSTEMS | |
| CA2125045A1 (en) | Nucleotide sequences coding an endopolygalacturonase inhibitor | |
| WO2001007589A2 (en) | Dna sequences encoding lipopeptide synthases | |
| Franche et al. | Characterization of several tropical strains of Anabaena and Nostoc: morphological and physiological properties, and plasmid content | |
| WO2001007588A2 (en) | Dna sequences which code nodulation factors | |
| WO2002024904A2 (en) | Method for producing ribonucleic acid (rna) | |
| WO2002029016A2 (en) | Thermostable polymerase based on thermococcus pacificus | |
| WO2001007587A2 (en) | Dna sequences coding for iron and heavy metal transporters | |
| WO2001007629A2 (en) | Dna sequences which code oygenases | |
| WO2001007591A2 (en) | Dna sequences which code export systems | |
| Patyka et al. | TRFLP profile of the assemblage of prokaryotic microorganisms in podzolic soils | |
| DE10225380B4 (en) | Production and secretion of glucosyltransferases | |
| DE2755171C3 (en) | Process for the production of proteins from methanol using methylotrophic microorganisms |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| AK | Designated states |
Kind code of ref document: A2 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW |
|
| AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
| DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
| AK | Designated states |
Kind code of ref document: A3 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW |
|
| AL | Designated countries for regional patents |
Kind code of ref document: A3 Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
| REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
| 122 | Ep: pct application non-entry in european phase | ||
| NENP | Non-entry into the national phase |
Ref country code: JP |