EP0288547A1 - Transformation de plantes - Google Patents

Transformation de plantes

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Publication number
EP0288547A1
EP0288547A1 EP87907654A EP87907654A EP0288547A1 EP 0288547 A1 EP0288547 A1 EP 0288547A1 EP 87907654 A EP87907654 A EP 87907654A EP 87907654 A EP87907654 A EP 87907654A EP 0288547 A1 EP0288547 A1 EP 0288547A1
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EP
European Patent Office
Prior art keywords
dna
plant
plasmid
orit
sequence
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Withdrawn
Application number
EP87907654A
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German (de)
English (en)
Other versions
EP0288547A4 (en
Inventor
Vicky Buchanan-Wollaston
Frank S. Cannon
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.)
Biotechnica International Inc
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Biotechnica International Inc
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Publication date
Application filed by Biotechnica International Inc filed Critical Biotechnica International Inc
Publication of EP0288547A1 publication Critical patent/EP0288547A1/fr
Publication of EP0288547A4 publication Critical patent/EP0288547A4/en
Withdrawn legal-status Critical Current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
    • 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)
    • C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
    • C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
    • C12N15/8205—Agrobacterium mediated transformation

Definitions

  • heterologous DNA DNA (a gene or other DNA) which is non-plant derived, modified, synthetic, derived from a different plant strain or species, or derived from a different location in the plant's genome.
  • bacterial plasmids are known to carry genetic sequences that enable transfer of plasmids from one bacterial cell to another, including the tra, mob and oriT loci.
  • the tra gene cluster is responsible for cell-to-cell contact and the formation of a mating bridge for the transfer of DNA.
  • the mob DNA sequence encodes proteins which nick and unwind plasmid DNA to enable its replicative transfer. These proteins specifically recognize oriT nucleotide sequences as their target, with specific protein molecules recognizing specific oriT sequences (See Guiney, D.G. et al., (1985) The Origin of Plasmid DNA Transfer During Bacterial Conjugation; In: Helinski, D.R., et al. (eds) "Plasmids in Bacteria", Plenum Press, New York, pp. 521-534).
  • DNA sequences which promote the transfer of DNA from one bacterial cell to another in bacterial (particularly Gram-negative) conjugation can also bring about the transfer of DNA (particularly desired heterologous genes) from bacterial cells to plant cells, in a manner analogous to known techniques involving the T-DNA (particularly the Ti border DNA) of A. tumefaciens.
  • Our discovery makes possible the production of a wide range of new and useful plants. For example, the discovery may make possible the transformation of plant cells without the need to rely on A. tumefaciens, and could thus open up the possibility of transforming even plant cells which cannot be infected by A.
  • tumefaciens e.g., certain monocotyledenous plants such as corn and other cereal grains, and certain dicotyledenous plants as well.
  • Our discovery also makes possible the transformation of A. tumefaciens - infectible plants using vectors which contain oriT and mob functions, but not Ti border DNA.
  • self-mobilizable broad host range plasmids such as RSF1010, which contain oriT and mob DNA sequences but no Ti border sequences, can be used in conjunction with A. tumefaciens to transform any plant which can be infected by A. tumefaciens.
  • the invention features, in general, a plant containing an engineered DNA construct including an oriT DNA sequence and heterologous DNA.
  • the DNA construct is integrated into a chromosome of the plant, and preferably the construct, in the plant, contains, in sequential order, a first portion of the oriT DNA sequence, the heterologous DNA, and the balance of the oriT DNA sequence.
  • the construct, in the plant further contains a mob DNA sequence between the two portions of the oriT DNA sequence.
  • Plants of the invention can be made by transforming a plant cell, under transforming conditions, with a first plasmid containing heterologous DNA and an oriT DNA sequence, and with a mob DNA sequence compatible with the oriT sequence, the mob DNA sequence being carried either on the first plasmid or on a second plasmid, neither the first nor the second plasmid including any DNA which is derived from or identical to Ti border DNA or A. tumefaciens.
  • a plant cell "transformed" with a heterologous gene means a plant cell in which the gene has gotten into the plant cell and is expressed and stably maintained in that cell (preferably, but not necessarily, by integration into a chromosome of the plant).
  • OriT and mob sequences are "compatible", as that term is used herein, where proteins encoded by the mob sequence recognize the oriT site (generally about 100-500 base pairs in length) and are capable of nicking the plasmid DNA bearing the heterologous gene and the oriT sequence to effect linearization of the plasmid DNA for its transfer into the plant cell.
  • the compatible mob sequence can be on the same plasmid as the oriT sequence and the heterologous gene (which must be on the same plasmid), or on another plasmid.
  • the plant is a monocotyledenous or dicotyledenous plant; and the heterologous DNA includes a gene selected from the group consisting of non-plant genes, modified genes, synthetic genes, and genes from a different plant strain or species.
  • the plasmid DNA is carried in a bacterial cell, preferably one of a species which is capable of growing in the rhizoplane or rhizointerstices of the plant species from which the plant cell is derived.
  • Bacteria growing in the rhizoplane are those which are in direct contact with the roots and root hairs of the plant, while bacteria growing in the rhizointerstices are those growing inside the roots or root hairs, in the interstices between cells.
  • a suitable host bacterial strain is potentially available for any plant which is associated with bacteria growing in the plant's rhizoplane or rhizointerstices, and such bacteria are known for virtually all plant species.
  • Rhizobium meliloti is a bacterial species known to associate closely with alfalfa; Rhizobium japonicum with soybeans; and Azospirillum brasilense with corn.
  • Agrobacterium tumefaciens is known to associate with a number of dicotyledenous plant species.
  • Such bacteria are desired carriers of the plasmid DNA because they can contain vir gene clusters capable of causing the bacteria to recognize the presence of plants (probably by virtue of plant secretions), which recognition is a necessary first step in transformation; tra genes, which are needed for conjugation of bacterial cells, are not needed, since it is plant transformation, not bacterial conjugation, which is occurring. Bacteria which are closely associated with a plant are also advantageously likely to contain other elements which aid in the recognition and transformation of plants.
  • the bacterial species chosen should be not only one which is associated with the plant, but also should contain vir genes; if it does not contain vir genes, the DNA used to transform it must include, as well as oriT and mob DNA, vir genes derived, e.g., from the A. tumefaciens Ti plasmid according to conventional techniques. (See Stachel and Nester (1986) EMBO J. 5: 1445-1454 for a restriction map of the vir gene cluster.) As an example of the necessity of vir genes, we have found that some vir- strains of A.
  • Rhizobium meliloti is a plant-associated bacterial species, yet we have found that R. meliloti cannot serve as a host for the transformation of plants with a plasmid including a heterologous gene and oriT and mob functions, at least in part because R. meliloti lacks vir genes.
  • the plasmid DNA also includes a DNA sequence encoding a selectable marker protein, and the method includes the step of selecting transformants on the basis of the presence of the selectable marker protein (which can itself be the desired product of the heterologous gene).
  • FIGS. 1, 7, and 8 are diagrammatic representations of plasmids of the invention.
  • Fig. 2 is a diagrammatic representation of an intermediate gene fusion used in the construction of the plasmids of Figs. 1, 7, and 8.
  • Figs. 3 and 5 are diagrammatic representations of plasmids containing synthetic Ti border DNA.
  • Fig. 4 is a diagrammatic representation of a plasmid containing neither Ti DNA nor mob and oriT sequences.
  • Fig. 6 is a diagrammatic representation of the region of pSUP104 containing the oriT and mob loci. Plasmid Components
  • the plasmids of the invention contain various DNA regions and sites, now discussed in greater detail.
  • plasmids of the invention preferably contain a DNA region which encodes a selectable marker protein for identification of transformants.
  • This marker protein can be any protein which can be expressed in plant cells and which enables phenotypic identification of plant cells which express the protein.
  • Preferred marker proteins are proteins which provide resistance to one or more antibiotics; currently most preferred is the protein aminoglycoside phosphotransferase, which inactivates antibiotics such as kanamycin neomycin and G418; transformants are those plant cells able to grow in the presence of antibiotic.
  • chloramphenicol acetyl transferase which provides resistance to chloramphenicol
  • dihydrofolate reductase which provides resistance to methotrexate
  • protein hygromycin-phosphotransferase encoded by the aph IV gene which confers resistance to the antibiotic hygromycin B.
  • Herbicide resistance can also be used as the selectable marker. As is mentioned above, herbicide resistance can constitute both the selectable marker and the desired function itself, in which case no additional heterologous gene need be inserted into the vector.
  • any protein which induces herbicide resistance can be produced, by expression of the introduced gene, in all of the plant tissues, including the herbicide-vulnerable leaves.
  • This foliar expression permits herbicide application at any time during crop growth, including the period after the appearance of leaves on the crop plant, and thus will be particularly useful in conferring resistance to post-emergent herbicides.
  • herbicides to which resistance can be induced include 2,2-dichloropropionic acid ("Dalapon”), glyphosate (N-phospho-m ⁇ thyl glycine; “Roundup”), cyanazine, chlorsulfuron, L-phosphinothricin, atrazine, the imidazolines, and 6-chloro-N-ethyl-N-(1-methyl)- 1,3,5-triazine-2,4-diamine (“Gesaprim 500L”).
  • Dalapon 2,2-dichloropropionic acid
  • glyphosate N-phospho-m ⁇ thyl glycine; "Roundup”
  • cyanazine chlorsulfuron
  • L-phosphinothricin atrazine
  • the imidazolines and 6-chloro-N-ethyl-N-(1-methyl)- 1,3,5-triazine-2,4-diamine
  • Mechanisms by which vector encoded protein-mediated herbicide resistance can be induced in plants according to the invention include the following: Detoxification - This involves either enzymatically altering the herbicide molecule by removing a substituent rendering the molecule non-toxic; cleaving the molecule; or adding a substituent to the molecule to render it non-toxic.
  • the first mechanism, removal of a substituent is the mechanism by which a bacterial dehalogenase inactivates, by removal of the chlorine atoms, the herbicide Dalapon.
  • the bacterial plasmid carrying the gene for dehalogenase has been isolated and is described in Beeching et al. (1983) J. Gen. Microbiol. 129, 2071.
  • An example of the addition of a substituent to detoxify a substance toxic to plants is the enzymatic addition of a phosphate group to kanamycin by aminoglycoside phosphotransferase, described above.
  • Herbicide Target - This involves introducing into the plant a mutant gene which codes for a form of the herbicide-targeted enzyme which is not susceptible to poisoning by the herbicide.
  • Increase in Target - This involves introducing into the plant a gene encoding the enzyme poisoned by the herbicide, to increase the amount of the enzyme produced by the plant.
  • Various bacteria are able to use the detoxification mechanism to render themselves resistant to herbicides. These bacteria are a convenient source of herbicide resistance-conferring genes. Such genes are generally isolated from such bacteria (generally soil bacteria) by first selecting bacteria either able to grow on the herbicide as a nutrient source or able to detoxify the herbicide. The enzymes responsible for herbicide detoxification in these bacteria are characterized and the genes encoding the enzymes are then isolated according to standard methods.
  • Eukaryotic (e.g., plant) messenger RNA's must be polyadenylated for efficient translation and processing.
  • Polyadenylati ⁇ n requires a recognition site for polyadenylation enzymes near the 3' end of the DNA region encoding the selectable marker.
  • DNA Sequences for Plasmid Transfer DNA derived from or substantially identical to (i.e., structurally similar enough to function in the same way) the mob and oriT loci of a self-mobilizable or self-transmissible bacterial plasmid (the latter contain tra genes and the former do not) is used to effect the transfer of the plasmids of the invention into plant cells.
  • the role of these loci in conjugative plasmid DNA transfer particularly from one Gram-negative bacterium to another, is well known.
  • the oriT-mob system is useful for the transfer of DNA from bacteria into plant cells as well.
  • mob and oriT sequences include those of the P-type, Q-type and W-type incompatability groups. Proteins encoded by the mob of one plasmid generally will only recognize the oriT site of that or a closely related plasmid, hence compatible mob and oriT sequences should be used together.
  • a preferred source of mob and oriT sequences is the Q-type incompatability group of plasmids, e.g., RSF1010 and its derivatives. These sequences may also be derived from P-type plasmids, e.g., RK2. Other plasmids from which the compatible mob and oriT sequences can be obtained are the hundreds of other known large self-transmissible plasmids.
  • these plasmids are too large to be used in toto and to be useful in the invention, either the mob and oriT sequences are removed, by conventional methods, or extraneous DNA is removed, leaving a smaller plasmid containing oriT and mob.
  • Nucleotide sequences of the oriT loci of several plasmids are given in Willetts and Wilkins (1984) Microb. Rev. 48:24-41.
  • Regulatory Sequences Transcription of both the selectable marker gene and the desired heterologous gene (if different from the marker gene), in order to lead to efficient expression, is preferably under the control of regulatory sequences normally expressed in plant cells, e.g., promoters such as those for the nopaline synthase (NOS) or octopine synthase (OS) genes of the T-DNA of Agrobacterium tumefaciens.
  • NOS nopaline synthase
  • OS octopine synthase
  • promoter sequences examples include those of the ribulose biphosphate carboxylase small subunit gene, the nitrate reductase gene, the glutamine synthase gene, and the 35S and 19S promoters of Cauliflower Mosaic Virus ("CAMV"). Synthetic, engineered, or altered natural promoters can also be used. In some instances, it will be desirable to use promoters which are regulated, e.g., promoters active only at certain times in the plant's development. It is desirable that the plasmids of the invention contain less than the entire structural gene normally under control of each promoter used, to ensure that metabolic energy of the transformed plant is not wasted producing protein encoded by the structural gene.
  • CAMV Cauliflower Mosaic Virus
  • the site for the insertion of a desired heterologous gene can be any site at which an endonuclease can act to cut the plasmid for insertion of the desired heterologous gene.
  • the site is unique in the plasmid, so that the endonuclease cuts the plasmid only in the desired location.
  • the desired heterologous gene can be any gene which is capable of being expressed in the plant, and which encodes a protein which enhances a beneficial feature of the plant, or provides a new beneficial feature.
  • examples of such genes are those encoding resistance to herbicides, resistance to diseases such as tomato fusarium wilt, proteins which can be produced to improve the protein content or other nutritive value of the plant, and genes encoding biopesticides.
  • the easiest modification to carry out is one involving a biochemical function which is controlled by a single protein, and is thus encoded by a single gene, more than one heterologous gene can be introduced, and the expression of such multiple genes controlled in a coordinate manner so as to introduce more complex biochemical functions into plants.
  • transcription of the desired heterologous gene is preferably under the control of a regulatory sequence normally expressed in plant cells.
  • a regulatory sequence normally expressed in plant cells Also, as mentioned above, there need be no additional heterologous gene if the marker itself has utility; e.g., if it confers herbicide resistance on the plant.
  • the fusion of the heterologous gene to the regulatory sequence can be carried out prior to the insertion of the heterologous gene into the vector, using conventional techniques.
  • the gene can be inserted into the vector by itself, and the regulatory sequence inserted upstream from the gene separately, prior to or following the insertion of the gene, using conventional methods.
  • plasmid pJP181 of the invention contains the aminoglycoside phosphotransferase gene from the transposon Tn5, encoding resistance to the antibiotics kanamycin and G418 (the kan r gene), under the transcriptional control of the NOS promoter from the Agrobacterium tumefaciens Ti plasmid; a polyadenylation site; the oriT and mob sequences from the broad host-range plasmid RSFIOIO (Scholz, P., et al., (1985) Replication Determinants of the Broad Host-Range Plasmid RSF1010, in Helinski, E.R. et al. (eds.).
  • Plasmids in Bacteria Plenum Press, New York, pp. 243-259.
  • Plasmid pJP181 was constructed as follows, beginning with the construction of a fusion of the kan r gene to the NOS promoter and polyadenylation site.
  • This fusion was derived from plasmid pVW104, the construction of which is described in Buchanan-Wollaston et al., "Plant Transformation Vector", U.S.S.N. 845,547, assigned to the same assignee as the present application, and hereby incorporated by reference.
  • the fusion can also be derived from plasmid pVW125 (ATCC No. 39929, also described in
  • the fusion allows expression of the kan r gene in plant cells, for use as a selectable marker.
  • the Bcll-Bglll fragment of pVW104 (Fig. 2), containing the kan r gene and polyadenylation site attached to the NOS promoter, was cloned into the BamHI site of pSUP104 (a plasmid derived from RSF1010) to yield plasmid pJP181 (Fig. 1). Plasmid pSUP104 is described in Simon et al.
  • pJP181 is similar in construction to pVW144 (Fig. 3), described in Buchanan-Wollaston, id, except that pVW144, unlike pJP181, contains a single copy of a 25-bp synthetic border sequence derived from A. tumefaciens Ti DNA, which is capable of effecting transformation of plant cells.
  • pJP181 When used in the "binary" transformation method described below, we found that pJP181, like pVW144, was able to transform tobacco cells to kanamycin resistance despite the absence in pJP181 of natural or synthetic DNA derived from transforming DNA of A. tumefaciens.
  • analogous vectors were constructed using, rather than pSUP104, a different plasmid, pRK290 (ATCC No. 37168; which is derived from RK2, ATCC No. 37125) as the backbone.
  • the Bcll-Bglll fragment of pVW104 and corresponding BamHI fragment of pVW144 were separately inserted into the Bglll site of pRK290, to create plasmids pVW170 (Fig. 4) and pVW171 (Fig. 5), respectively.
  • pVW170 and pVW171 were used in a "binary" transformation, pVW171 was, by virtue of the synthetic border sequence, able to transform tobacco cells, but pVW170 was not.
  • the oriT and mob loci of RSF1010 are known to be contained on a 2.9 kb Aval fragment, which also contains the oriV sequence (the vegetative origin of replication) and part of the repB gene (see Fig. 6).
  • This 2.9 kb fragment was cloned into the EcoRI site of pVW170 to yield pVW210 (Fig. 7) and pVW214 (Fig. 8), differing only in the orientation of the insert.
  • pVW206 in which the hyg R gene is under the control of the NOS promoter, was constructed by cloning the Bcll-Sall fragment of pVW189 (described in Buchanan-Wollaston et al., id) into pSUP104. Plasmid pVW206 was found to be capable of transforming plant cells by the "binary" method described below.
  • Plasmids pJP194 and pJP195 were found to be capable of transforming plant cells by the "binary" method described below. Additional vectors were constructed utitilizing the CAMV 19S promoter (strain CM1841); the sequence of the CAMV 19S promoter is given in Table I, below. A 129 bp fragment containing this promoter was isolated from plasmid pBL101 by first removing a 0.5 kb EcoRI fragment and then cutting with Mnll.
  • pJP304 which contains a fusion of the kan R gene to the 19S promoter and a polydenylation site
  • pJP305 which contains a fusion of the hyg R gene to the 19S promoter and a polyadenylation site. Both these fusions have a pSUP104 backbone, and lack Ti border sequences.
  • Plasmid pJP181 has a unique HindIII site and Sall site and plasmid pVW210 has a unique Bglll site into which a desired heterologous gene can be inserted, using conventional techniques. Similar sites exist, or can be created by standard techniques, on plasmids pVW206, pJP194, pJP195, pJP304, and pJP305.
  • Plant Transformation The plasmids of the invention can be used to transform plant protoplasts or non-protoplast cells, using a "binary" cocultivation technique. The following is an example of this technique, using A. tumefaciens as the bacterial delivery system.
  • Plasmids containing oriT and mob genes (pJP181, pVW210, pVW206, pJP194, and pJP195) were transferred to an A. tumefaciens strain, e.g., LBA4404, which carries a Ti plasmid, pLBA4404, deleted of the T-DNA region, so that cocultivation does not result in plant tumor formation.
  • LBA4404 binary cocultivation, are described in Hoekema et al.
  • pLBA4404 does, however, retain the native Ti vir (for virulence) functions, at least some of which are essential for the transfer of a hybrid plasmid of the invention from A. tumefaciens LBA4404 to the host plant cell.
  • the A. tumefaciens containing the hybrid plasmid was then cocultivated with plant cells to insert the hybrid plasmid into the plant cells, where the plasmid DNA integrated into the plant cell chromosome. Plant transformation was accomplished using the "quick dip" method, which involves the dipping of plant explants in a bacterial (e.g., A .
  • the plant cells are cultured under conditions effecting the regeneration of mature plants.
  • Such methods are known, e.g., for the regeneration of tobacco plants from callus culture. See, for example, "Plant Tissue Culture: Methods and Applications in Agriculture", Trevor A. Thorpe, ed., Academic Press, New York, 1981; particularly the article by O.L. Gamborg and J.P. Shyluk at pp. 21-24, and the chart at page 33.
  • transforming plasmid DNA is integrated into a plant cell chromosome such that the chromosome has integrated into it the heterologous DNA flanked by two portions of the oriT sequence. From studies of bacterial conjugation, it is well known that oriT sequences are nicked before transfer of plasmid DNA from one cell into another, and that the transfer of DNA is initiated at the oriT sequence.
  • the nicking site or the oriT sequence of RSF1010 has been localized to a 153bp sequence of RSF1010 (Willetts and Wilkins, id.) and to an 80 bp sequence within that 153 bp sequence (Derbyshire et al. M.G.G., 206:154, 1987).
  • the engineered vectors of our invention are nicked at the oriT site prior to transfer into a plant cell, which results in a linear DNA molecule which includes a portion of the oriT DNA sequence adjacent to heterologous DNA followed by the balance of the oriT DNA sequence. It is this sequence which, we believe, is integrated into the host plant genome following transformation.
  • Other Embodiments are within the following claims.
  • oriT and mob sequences may be obtained from other self-mobilizable plasmids, for example, the plasmids RK2 and pPH1.
  • the sequence of the RK2 oriT region is given in Willetts and Wilkins, id., and the RK2 mob genes are located in one of the tra clusters (Guiney, id).
  • Other promoter sequences can be used, e.g., those described in Buchanan-Wollaston et al., id.

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Abstract

Plante contenant une structure d'ADN manipulée génétiquement comprenant une séquence d'ADN oriT et un ADN hétérologue.
EP19870907654 1986-11-03 1987-11-03 Plant transformation Withdrawn EP0288547A4 (en)

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US92662986A 1986-11-03 1986-11-03
US926629 1992-08-03

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EP0288547A1 true EP0288547A1 (fr) 1988-11-02
EP0288547A4 EP0288547A4 (en) 1991-03-20

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0267159A3 (fr) * 1986-11-07 1990-05-02 Ciba-Geigy Ag Procédé de modification génétique de monocotylédones
AU4946700A (en) * 1999-06-02 2000-12-28 Yissum Research Development Company Of The Hebrew University Of Jerusalem Method for conferring resistance to crown gall disease
EP1130105A1 (fr) * 2000-03-01 2001-09-05 Rijksuniversiteit te Leiden Transformation des cellules eucaryotes par des vecteurs transposables

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1986003516A1 (fr) * 1984-12-13 1986-06-19 Biotechnica International, Inc. Vecteur de transformation vegetale
EP0218571A2 (fr) * 1985-08-07 1987-04-15 Monsanto Company Plantes résistant au glyphosate

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NL8300699A (nl) * 1983-02-24 1984-09-17 Univ Leiden Werkwijze voor het inbouwen van vreemd dna in het genoom van tweezaadlobbige planten; werkwijze voor het produceren van agrobacterium tumefaciens bacterien; stabiele cointegraat plasmiden; planten en plantecellen met gewijzigde genetische eigenschappen; werkwijze voor het bereiden van chemische en/of farmaceutische produkten.
US4686184A (en) * 1983-07-01 1987-08-11 Lubrizol Genetics, Inc. Gene transfer vector
US4626504A (en) * 1983-07-01 1986-12-02 Lubrizol Genetics, Inc. DNA transfer vector for gram-negative bacteria
EP0429093B1 (fr) * 1984-05-11 2001-08-08 Syngenta Participations AG Transformation du génotype de plantes

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1986003516A1 (fr) * 1984-12-13 1986-06-19 Biotechnica International, Inc. Vecteur de transformation vegetale
EP0218571A2 (fr) * 1985-08-07 1987-04-15 Monsanto Company Plantes résistant au glyphosate

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
PROC. NATL. ACAD. SCI. (USA), vol. 83, June 1986, pages 3895-3899; G.C. JEN et al.: "The right border region of pTiT37 T-DNA is intrinsically more active than the left border region in promoting T-DNA transformation" *
PROC. NATL. ACAD. SCI. (USA), vol. 83, June 1986, pages 4428-4432; R.B. HORSCH et al.: "Rapid assay of foreign gene expression in leaf discs transformed by Agrobacterium tumefaciens: Role of T-DNA borders in the transfer process" *
SCIENCE, vol. 233, 25th July 1986, pages 478-481; D.M. SHAH et al.: "Engineering herbicide tolerance in transgenic plants" *
See also references of WO8803564A1 *

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WO1988003564A1 (fr) 1988-05-19
AU604805B2 (en) 1991-01-03
JPH01501598A (ja) 1989-06-08
EP0288547A4 (en) 1991-03-20
AU8240087A (en) 1988-06-01

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