EP0927246A2 - Adenylosuccinate-synthetase - Google Patents

Adenylosuccinate-synthetase

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Publication number
EP0927246A2
EP0927246A2 EP97943832A EP97943832A EP0927246A2 EP 0927246 A2 EP0927246 A2 EP 0927246A2 EP 97943832 A EP97943832 A EP 97943832A EP 97943832 A EP97943832 A EP 97943832A EP 0927246 A2 EP0927246 A2 EP 0927246A2
Authority
EP
European Patent Office
Prior art keywords
nucleic acid
expression
plant
plants
expression cassette
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.)
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Application number
EP97943832A
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German (de)
English (en)
Inventor
Jens Lerchl
Ralf-Michael Schmidt
Helmut Schiffer
Uwe Sonnewald
Ralf Badur
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BASF SE
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BASF SE
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Filing date
Publication date
Application filed by BASF SE filed Critical BASF SE
Publication of EP0927246A2 publication Critical patent/EP0927246A2/fr
Withdrawn legal-status Critical Current

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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
    • C12N9/93—Ligases (6)
    • 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
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance

Definitions

  • the present invention relates to expression cassettes, coding for non-vegetable adenylosuccinate synthetases (ADSS), which impart resistance to plant ADSS inhibitors to plants; Vectors and microorganisms containing such expression cassettes; transgenic plants transformed therewith; the corresponding expression products and nucleic acid sequences; and an expression kit for use in transforming a plant host.
  • ADSS non-vegetable adenylosuccinate synthetases
  • plants are able to synthesize their cell components from carbon dioxide, water and inorganic salts. This process is only possible by using biochemical reactions to build up organic substances. In particular, plants must also synthesize the nucleotides as components of the nucleic acids de novo.
  • the enzyme reactions of purine biosynthesis can start from
  • Pliosphoribosyl pyrophosphate can be divided into the following steps:
  • step c) The reaction sequence of step c) is shown schematically in the attached FIG. 1.
  • ADSS occupies a special position. The enzyme catalyzes the following reaction:
  • ADSS was purified from E. coli for homogeneity (Bass et al., 1987, Aren. Biochem. Biophys., 256, 335-342), the crystal structure was determined (Poland et al., 1993, J. Biol. Che. , 268, 25334-25342) and kinetic properties of the enzyme 5 were characterized (Kang and Fromm, 1995, J. Biol. Chem., 270,
  • the enzyme was also purified from Dictyostelium diseoideum (Jahngen and Rossomando, 1984, Arch. Biochem. Biophys., 229, 145-154) and rabbit muscle (J. Biol. Chem., 1974, 249 (2), 459-464). Vegetable adenylosuccinate synthetase 10 was obtained from wheat seedlings (Hatch, 1967, Phytochemistry, 6,
  • the adenylosuccinate synthetase is present in E. coli as a dimer of two 15 48 kD polypeptides. No data are available for herbal ADSS.
  • ADSS ADSS Genes coding for ADSS have so far been derived from Escherichia coli (EMB access number J04199), Bacillus subtilis (J02732),
  • ADSS activity No cofactors are required to measure ADSS activity.
  • the active substances hadacidin and alanosine (Stayton et al., 1983, Curr. Top. Cell. Regul., 22, 103-141) as well as hydantoine or its metabolite 5 '-phosphohydantocidine have been described (Siehl et al., 1996, Plant Physiol., 110, 753-758), which inhibit the enzyme activity of plant ADSS.
  • the phytotoxic active ingredient hydantocidin was isolated from Streptomyces 40 hygroscopicus (strain SANK 63584) (Nakajima et al., 1991, J. Antibiot., 44, 293-300). It has been shown that 5'-phosphohydantoeidine as the plant metabolite is the actual inhibitor of ADSS (Siehl et al., 1996, Plant Physiol., 110, 753-758). Hydantocidin exerts its toxic effect only in plants, has a low mammalian toxicity and does not act on various microorganisms investigated (Nakajima et al., 1991, J. Antibiot., 44, 293-300). It would be desirable, if ways could be found to specifically change the response of plants to ADSS inhibitors.
  • the invention is therefore based on the object of providing means by means of which the response of plants to ADSS inhibitors can be specifically modified.
  • this modification should be feasible by genetic engineering.
  • This object is achieved by providing an expression cassette containing regulatory under genetic control
  • Nucleic acid sequences the coding nucleic acid sequence for a protein which imparts resistance to inhibitors of plant adenylosuccinate synthetase to a plant host.
  • FIG. 1 shows the de novo purine biosynthesis in plants
  • FIG. 2 shows the nucleic acid sequence of the adenylosuccinate synthase from Escherichia coli, including the 5 'and 3' terminal Ba HI cleavage sites
  • FIG. 3 shows the amino acid sequence of the adenylosuccinate synthetase from Escherichia coli
  • FIG. 4 oligonucleotide sequences for isolating the nucleic acid sequence of the adenylosuccinate synthetase from Escherichia coli
  • FIG. 5 shows the nucleic acid sequence of the transit peptide of chloroplast plastid transketolase in three reading frames
  • FIG. 6 shows the structure of the expression cassettes and transformation vectors pTP09, pTPlO and pTPll;
  • FIG. 7 shows the structure of the expression cassettes and the transformation vector pTP09-ASS.
  • a first object of the present invention relates to expression cassettes which are suitable for transforming a plant host and which contain a coding sequence which give the host resistance to inhibitors of plant ADSS.
  • resistance means the artificially acquired resistance to the action of plant ADSS inhibitors. It includes the partial and, in particular, the complete insensitivity to these inhibitors for at least one generation of plants.
  • expression cassettes for a plant host are selected from whole plants, plant cells, plant tissues or plant parts, such as leaves, roots, fruits.
  • the plant host is selected from crop plants, cells, tissues or parts derived therefrom.
  • suitable crop plants can be mentioned: cereals, corn, soybeans, rice, cotton, sugar beet, canola, sunflower, flax, potato, tobacco, tomato, rapeseed, alfalfa, lettuce and the various tree, nut and Wine species.
  • the present invention offers the advantage that, after induction of a selective resistance of the crop plant to plant ADSS inhibitors, these inhibitors can be used as specific herbicides against non-resistant plants.
  • Non-limiting examples of inhibitors of this type include alanosine, hadacidin, hydantocidin and metabolites and functionally equivalent derivatives thereof.
  • Functionally equivalent derivatives of plant ADSS inhibitors have a comparable spectrum of action as the specifically named substances, with lower, equal or higher inhibitory activity (e.g. expressed in g inhibitor per hectare of cultivated area, required to completely suppress the growth of non-resistant plants).
  • the invention relates in particular to expression cassettes whose coding sequence contains a non-vegetable ADSS nucleic acid sequence which is tolerant to plant ADSS inhibitors or a functional equivalent thereof.
  • the ADSS nucleic acid sequence can e.g. be a DNA or a cDNA sequence.
  • Coding sequences suitable for insertion into an expression cassette according to the invention are, for example, those which essentially comprise a microbial DNA sequence which are suitable for an adenylosuccinate synthetase from microorganisms of the genera Escherichia, Bacillus, Haemophilus, Dictyostelium, Saccharomyces or Schizosaccharomyces and in particular from Escherichia coli subtilis, Haemophilus influenzae, Dictyostelium discoideum, Saccharomyces pombe or Schizosaccharomyces pombe.
  • Particularly suitable is an ADSS sequence which essentially corresponds to the DNA sequence from E. coli shown in FIG. 2 (SEQ ID N0: 1).
  • artificial DNA sequences are suitable as long as they induce the desired resistance as described above.
  • Such artificial DNA sequences can be determined, for example, by back-translation of proteins constructed by means of molecular modeling which have adenylosuccinate synthetase activity or by in vitro selection.
  • ADSS coding DNA sequences obtained by back-translating an ADSS polypeptide sequence according to the codon usage specific for the host plant.
  • the specific codon usage can easily be determined by a person skilled in plant genetic methods by computer evaluations of other, known genes of the plant to be transformed.
  • the invention also relates to the sequences which are functionally equivalent to the above nucleic acid sequences and to the amino acid sequences derived therefrom.
  • functional equivalents are sequences which essentially correspond to one another. This means in particular sequence variants which have arisen as a result of natural or artificial mutations in an ADSS sequence, provided that the desired ADSS activity, which is necessary to maintain plant metabolism, is retained.
  • the exogenous ADSS activity transformed in the plant host can therefore be higher, comparable or somewhat lower than that of the endogenous plant ADSS. Mutations include substitutions, deletions, exchanges or insertions of one or more nucleotide or amino acid residues.
  • Substitutions mean exchanges of nucleotides or amino acids. So-called silent or conservative exchanges are preferred. A silent nucleotide exchange does not change the amino acid sequence. A conservative exchange results in an amino acid substitution by an amino acid residue with comparable properties, e.g. Size, charge, polarity, solubility. Examples of amino acid pairs with similar properties are Glu and Asp, Val and Ile, Ser and Thr.
  • a deletion means the removal of at least one nucleotide or at least one amino acid residue.
  • Preferred positions for deletions are the DNA regions which code for the termini of the polypeptide and the links between the individual protein domains. According to the invention, particular preference is given to DNA sequences which code for ADSS proteins and which are reduced by N-terminal truncations by up to about 100, e.g. by 20 to 100 amino acids, from the sequence shown in FIG. 3 (SEQ ID N0: 2).
  • insertions comprise insertions of at least one nucleotide or amino acid residue into one of the sequences according to the invention.
  • Sequences which code for fusion proteins are to be mentioned as further functionally equivalent nucleic acid sequences according to the invention, part of the fusion protein being a non-vegetable ADSS polypeptide or a functionally equivalent part thereof.
  • the second part of the fusion protein can be, for example, another polypeptide with the same or different enzymatic activity. However, this is preferably a regulatory protein sequence, such as a signal or transit peptide, which directs ADSS to the desired site of action.
  • the invention thus also relates to expression cassettes whose coding sequence codes for an ADSS fusion protein, part of the fusion protein being a transit peptide which controls the translocation of the ADSS sequence.
  • Chloroplast-specific transit peptides are particularly preferred which, after translocation of the ADSS sequence into the plant chloroplasts (main site of purine biosynthesis in plants), are enzymatically split off from the ADSS part.
  • the transit peptide is particularly preferably derived from plastid transketolase (TK) or a functional equivalent of this transit peptide.
  • TK plastid transketolase
  • An expression cassette which contains one of the transit peptide DNA sequences shown in FIG. 5 (SEQ ID NOs: 3,4,5) is particularly preferred.
  • an expression cassette according to the invention also comprise regulatory nucleic acid sequences which control the expression of the coding sequence in the host cell.
  • an expression cassette according to the invention comprises upstream, i.e. at the 5 'end, the coding sequence a promoter and downstream, i.e. at the 3 'end a polyadenylation signal, and optionally further regulatory elements which are operatively linked to the intervening coding sequence for ADSS and / or transit peptide.
  • An operative link is understood to mean the sequential arrangement of the said regulatory elements in such a way that each of the regulatory elements can fulfill its function in the expression of the coding sequence.
  • any promoter which can control the expression of foreign genes in plants is suitable as the promoter of the expression cassette according to the invention.
  • a plant promoter is preferably used in particular.
  • the 35S CaMV promoter from the Cauliflower mosaic virus (Franck et al. (1980) Cell 21, 285-294) is particularly preferred.
  • This promoter contains different recognition sequences for transcriptional effectors, which in their entirety lead to a constitutive expression of the introduced gene (Benfey et al. (1989) EMBO J. 8, 2195-2202).
  • Preferred polyadenylation signals are plant polyadenylation signals, preferably those which essentially correspond to T-DNA polyadenylation signals from Agrobacterium tumefaciens, in particular the polyadenylation signal of gene 3 of the T-DNA of the Ti plasmid pTiACH5.
  • An expression cassette according to the invention is produced by fusing a suitable promoter with a suitable ADSS-DNA and preferably a DNA coding for a chloroplast-specific transit peptide inserted between the promoter and ADSS-DNA, and a polyadenylation signal according to common recombination and cloning techniques, as described, for example, in T. Maniatis, EF Fritsch and J. Sambrook, Molecular Cloning: A Laboratory manual, Cold Spring Harbor laboratory, Cold Spring Harbor, NY (1989) and in T.J. Silhavy, M.L. Berman and L.W. Enquist, Experiments with Gene Fusions, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1984) and in Ausubel, F.M. et al. , Current Protocols in Molecular Biology, Greene Publishing Assoc. and Wiley-Interscience (1987).
  • ADSS-DNA or cDNA required for the production of expression cassettes according to the invention is preferably obtained with the aid of the
  • PCR Polymerase chain reaction
  • the invention also relates to an expression kit which comprises at least three expression cassettes, at least two of which contain a variable region or frame shift sequence which differ from one another by insertion or deletion, preferably insertion, of a number of nucleotides which cannot be divided by 3 and thus shift the reading frame for a sequence inserted downstream by one or two nucleotides.
  • the 5 'end of the frame shift sequence is linked to the transit peptide sequence.
  • the frame shift sequence includes a restriction site at the 3 'end into which, for example, the ADSS-DNA sequence is inserted.
  • the provision of at least three vectors, each containing an expression cassette of the kit with different reading frames for the inserted gene, ensures that the fusion constructs consist of any DNA sequence and a DNA sequence coding for a transit peptide. can be expressed in three different reading frames, with at least one of the constructs leading to the expression of functional gene product (such as ADSS).
  • the construction diagrams for an expression kit according to the invention containing three expression cassettes and which is particularly suitable for the transformation of plants are shown in the attached FIG. 6.
  • the invention also relates to the use of such kits for transforming a plant host.
  • the expression cassettes according to the invention can be cloned into suitable vectors which enable their multiplication, for example in E. coli.
  • suitable cloning vectors include pBR332, pUC series, M13mp series and pACYC184.
  • Another object of the invention therefore relates to recombinant vectors, such as e.g. Plasmids or viruses containing at least one expression cassette according to the invention.
  • a particularly preferred recombinant plasmid called pTP09-ASS contains the gene construct shown in FIG. 7 and gives the plant host transformed therewith resistance to plant ADSS inhibitors, such as e.g. Hydantocidin.
  • an expression cassette according to the invention is inserted as an insert in a recombinant vector, the vector DNA of which contains additional functional regulatory signals, for example sequences for replication or integration.
  • Suitable vectors are inter alia in "Methods in Plant Molecular Biology and Biotechnology” (CRC Press), Chap. 6/7, p.71-119.
  • the vectors according to the invention can be used to transform plants, plant cells, plant tissues or parts.
  • the DNA constructs fused according to the invention can also be transferred into plant genomes by various other known methods. Suitable methods are, for example, protoplast transformation by polyethylene glycol-induced DNA uptake, electroporation, sonication or microinjection, and the transformation of intact cells or tissue by micro or macro injection into tissue or embryos, tissue electroporation, incubation of dry embryos in DNA-containing solution, biolistic shear gene transfer and particularly preferably Agrobacterium transform ation. The methods mentioned are described, for example, in B. Jenes et al. , Techniques for Gene Transfer; in Transgenic Plants, Vol. 1, Engineering and Utilization, edited by SD Kung and R. Wu, Academic Press, 1993, pp. 128-143 and in Potrykus (1991) Annu. Rev. Plant Physiol. Plant Molec. Biol. 42, 205-225).
  • the fused construct is preferably cloned into a vector, for example pBin19, which is suitable for transforming Agrobacterium tumefaciens.
  • Agrobacteria transformed with such a vector can then be used in a known manner to transform plants, in particular crop plants, such as e.g. of tobacco plants can be used, for example, by wounding leaves or leaf pieces in an agrobacterial solution and then cultivating them in suitable media.
  • the transformation of plants by agrobacteria is known, among other things, from F.F. White, Vectors for Gene Transfer in Higher Plants; in Transgenic Plants, Vol. 1, Engineering and Utilization, edited by S-d Kung and R. Wu, Academic Press, 1993, pp. 15-38 and from S.B.
  • Transgenic plants which express ADSS-DNA integrated into the expression cassette according to the invention can be regenerated in a known manner from the transformed cells of the wounded leaves or leaf pieces.
  • ADSS transgenically expressed ADSS
  • an enzyme assay as described in Baugher et al. , Biochem. Res. Commun., 94: 123-129 (1980); Stayton et al., Curr. Top. Cell. Regul. , 22: 103-141 (1983); and Bass et al., Arch. Biochem. Biophys., 256: 335-342 (1987).
  • the invention also relates to microorganisms, such as Bacteria or fungi which contain a recombinant vector according to the invention.
  • the invention also relates to transgenic plants transformed with a vector or microorganism according to the invention, and to transgenic cells, tissues, parts and propagation material of such plants.
  • Transgenic crop plants such as e.g. Cereals, corn, soybeans, rice, cotton, sugar beet, canola, sunflower, flax, potato, tobacco, tomato, rapeseed, alfalfa, lettuce and the various tree, nut and wine species.
  • transgenic plants, plant cells, tissue or parts can be treated with an active ingredient which inhibits the plant ADSS, as a result of which those which have not been successfully transformed Plants, cells, tissues or parts of plants die.
  • suitable active substances are alanosine, hadacidin and in particular hydantocidin as well as metabolites and functional derivatives of these compounds.
  • the ADSS-DNA inserted into the expression cassette according to the invention can thus be used as a selection marker.
  • Another object of the invention thus relates to the use of vectors or microorganisms transformed therewith for the transformation of plants, plant cells, plant tissues or plant parts, in particular for the expression of exogenous proteins, glycoproteins or fusion proteins.
  • the aim of the use is preferably to impart resistance to inhibitors of plant ADSS.
  • the invention also relates to the expression products produced according to the invention, in particular the fusion proteins from transit peptide and protein part with non-plant ADSS activity.
  • Example 1 PCR amplification of Escherichia coli adenylosuccinate synthetase using synthetic oligonucleotides
  • the PCR amplification of the Escherichia coli ADSS was carried out in a DNA thermal cycler from Perkin Elmer.
  • the oligonucleotides used (SEQ ID NO: 6, 7) are shown in FIG. 4 and were taken from the published sequence.
  • the reaction mixtures contained 8ng / ⁇ l genomic DNA from Escherichia coli, 0.5 ⁇ M of the corresponding oligonucleotides, 200 ⁇ M nucleotides (Pharmacia), 50 mM KCl, 10 M Tris-HCl (pH 8.3 at 25 ° C.), 1 , 5 mM MgCl) and 0.02 U / ⁇ l Taq polymerase (Perkin Elmer).
  • the amplification conditions were set as follows:
  • Annealing temperature 45 ° C, 1 min
  • nucleotides 11749-11939 derived from the octopine synthase (OCS) gene, was isolated as a PvuII-Hindlll fragment and after addition of Sphl linkers to the PvuII interface between the Sphl-Hindlll- Interfaces of the vector were cloned, resulting in the plasmid pBinAR (Höfgen and Willmitzer (1990) Plant Science 66, 221-230).
  • the cDNA sequence of the transit peptide (TP) of the transketolase (TK) was taken from the plasmid pBluescript TK-26 (DE-A-19501906) and with the aid of synthetic oligonucleotides as a Kpnl-BamHI fragment via a polymerase chain reaction into the plasmid pBinAR - riert.
  • three vectors were obtained as plant expression cassettes (pTP09, pTPlO, pTPll, see FIGS. 5 and 6), which allow chimeric gene constructs with the cDNA transit sequence of the plastid transketolase in three different reading frames to create.
  • Example 3 Creation of a plant expression cassette for the adenylosuccinate synthetase
  • the DNA fragment coding for the adenylosuccinate synthetase was cloned into the vector pTP09 as a BamHI fragment.
  • the plasmid pTP09-ASS resulted (see FIG. 7). Fusion of the transit peptide (TP / TK) with the adenylosuccinate synthetase ensured the import of the protein into the chloroplasts.
  • the plasmid pTP09-ASS was in Agrobacterium tumefaciens
  • C58C1 pGV2260 transformed.
  • the transformation of Agrobacterium tumefaciens was carried out according to the method of Höfgen and Willmitzer (Nucl. Acid Res. (1988) 16, 9877).
  • the agrobacteria were grown in YEB medium (Vervliet et al., J. Gen. Virol. (1975) 26, 33).
  • YEB medium Vervliet et al., J. Gen. Virol. (1975) 26, 33.
  • For the transformation of tobacco plants (Nicotiana tabacum cv. Samsun NN) a 1:50 dilution of an overnight culture of a positively transformed agrobacterial colony in Murashige-Skoog Medium ((1962) Physiol. Plant. 15, 473) with 2% sucrose (2MS- Medium) is used.
  • Leaf disks of sterile plants (each about 1 cm 2 were incubated in a Petri dish with a 1:50 agrobacterial dilution for 5 to 10 minutes. This was followed by a 2-day incubation in the dark at 25 ° C. on 2MS medium with 0.
  • RNA total RNA of tobacco plants, as in Logemann et al. (Anal. Biochem. (1987) 163,21).
  • 20 ⁇ g RNA were separated in a 1.5% agarose gel containing formaldehyde. After electrophoretic separation of the RNA molecules, the RNA was transferred to a nylon membrane by capillary transfer. The detection of specific transcripts was carried out as described in Amasino (Anal. Biochem. (1986) 152, 304).
  • the cDNA fragments used as a probe were radioactively labeled with a random primed DNA labeling kit (Boehringer, Mannheim).
  • Example 7 Enzyme assay of adenylosuccinate synthetase isolated from transgenic tobacco plants
  • the test mixture contained 14 mM Tris-HCl pH 8.3; 6 mM MgCl 2 ; 0.4 mM IMP; 0.1 mM GTP; 0.5 mM phosphoenol pyruvate; 0.1 mM ATP; 2 U / ml pyruvate kinase; 3 mM aspartate and in a 1 ml test batch 10 to 100 ⁇ l enzyme preparation.
  • the recombinant ADSS showed no inhibition by hydantocidin.
  • Example 8 Testing of hydantocidine-resistant tobacco plants
  • Tobacco plants transformed with the plasmid pTP09-ASS were grown in tissue culture on 2MS medium with 0.8% Bacto agar, 250 mg / 1 claforan, 50 mg / 1 kanamycin, and axial shoots were transferred to corresponding medium with 220 mg / 1 hydantocidin. Untransformed plants died within a few weeks, while resistant plants continued to grow.
  • ORGANISM Escherichia coli

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Abstract

L'invention concerne des cassettes d'expression, qui confèrent aux plantes, aux cellules végétales, aux tissus végétaux et parties de plantes, une résistance contre les inhibiteurs des adénylosuccinate-synthétases végétales. L'invention concerne en outre l'utilisation desdites cassettes d'expression dans des vecteurs appropriés pour transformer des plantes, des cellules végétales, des tissus végétaux et parties de plantes.
EP97943832A 1996-09-04 1997-09-04 Adenylosuccinate-synthetase Withdrawn EP0927246A2 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE19635917 1996-09-04
DE19635917 1996-09-04
PCT/EP1997/004812 WO1998010074A2 (fr) 1996-09-04 1997-09-04 Adenylosuccinate-synthetase

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EP0927246A2 true EP0927246A2 (fr) 1999-07-07

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JP (1) JP2001500008A (fr)
AU (1) AU741624B2 (fr)
BR (1) BR9711658A (fr)
CA (1) CA2264677A1 (fr)
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EP1222293A2 (fr) * 1999-10-01 2002-07-17 Basf Aktiengesellschaft Synthetase de gmp (guanosine monophosphate) provenant de vegetaux
EP1224292A2 (fr) * 1999-10-25 2002-07-24 Basf Aktiengesellschaft Synthase de formylglycinamidinribotide d'origine vegetale
US6871421B2 (en) 2001-09-21 2005-03-29 Daniel R. Potter Footwear with bladder type stabilizer
CN108018323B (zh) * 2018-01-12 2021-04-02 中国医学科学院药用植物研究所 一种腺苷酸基琥珀酸或盐的制备方法

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JPH01501038A (ja) * 1986-09-26 1989-04-13 カルジーン，インコーポレイティド 発現生成物の細胞内指図された輸送
GB8626862D0 (en) * 1986-11-11 1986-12-10 Jefferson R A Gene fusion
TW201794B (fr) * 1991-05-03 1993-03-11 American Cyanamid Co
US5688939A (en) * 1994-12-22 1997-11-18 Novartis Finance Corporation Plant adenylosuccinate synthetase and DNA coding therefor
DE19501906A1 (de) * 1995-01-23 1996-07-25 Basf Ag Transketolase

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Title
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AU741624B2 (en) 2001-12-06
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WO1998010074A2 (fr) 1998-03-12
BR9711658A (pt) 1999-08-24
JP2001500008A (ja) 2001-01-09
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AU4553097A (en) 1998-03-26

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