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TRANSFORMATION METHOD AND TRANSGENIC PLANTS PRODUCED THEREBY, P Christou & A Kohli (2005). Plant Bioscience Ltd., UK. Patent # US6846970.
This invention relates to methods for producing, at a high frequency, transgenic plants that contain little if any vector sequences, have simple integration patterns, contain few copies of the transgene at each locus, express the transgene at all stages of development and do not exhibit transgene silencing. The method comprises introducing minimal transgene expression cassettes, which are substantially or totally devoid of vector sequences, by direct DNA transfer, preferably by particle or microprojectile bombardment. This invention also relates to transformed plant cells, the transgenic plants regenerated therefrom, and subparts of the transgenic plants produced by the methods of this invention. The invention also includes all progeny and subsequent progeny (i.e., all subsequent generations) derived from primary transformants through selfing or crossing. |
A VEHICLE FOR PLANT TRANSFORMATION, S Kushnir, R D Maier & R Herrmann (2005). VIB vzw, Belgium, University of Ghent, Belgium & Ludwig Maximilians University, Germany. Patent # WO2005059146.
The invention relates to the field of transgenic plants. More specifically the invention relates to a recombinant plastid vehicle which can be used for transformation. In addition said vehicle can be used for the generation of inter-species hybrid plants and for the optimisation of plant fitness. |
PLASTID TRANSFORMATION USING MODULAR VECTORS, S Herz, H-U Koop, T J Golds & C Eibl (2005). Icon Genetics AG, Germany. Patent # EP1527185.
Generating transgenic plants, or plant cells, that are transformed in the plastome, is new. Generating transgenic plants, or plant cells, that are transformed in the plastome comprises first introducing two DNA molecules (A1, A2) into the plastid, where A1 has a first region homologous to a region of the plastome to direct integration, and a sequence of interest (I), and A2 contains a second region homologous to a region of the plastome to direct integration, and a second sequence of interest (II), with a segment of (II) being homologous with a segment of (I). Transformants are selected that contain an integration sequence (IS) integrated stably into the plastid, where IS contains at least parts of (I) and (II) as a continuous sequence. Independent claims are also included for: (1) a kit containing A1 and A2; (2) a DNA molecule (X) for plastid transformation comprising a sequence of interest and a region homologous to a region of the plastid, to direct integration; (3) a library of (X), where each molecule contains a different sequence of interest; (4) plants, and their cells, transformed with (X); and (5) plants, or their cells or seeds, produced by the new method. |
PLASTID TRANSFORMATION, E Boudreau, W Gu, A de Framond & P Heifetz (2004). Syngenta Participations AG, USA. Patent # US2004133937.
The present invention relates to the field of plastid transformation. The invention provides transformation vectors and methods to obtain transplastomic plants or algae having a transformed plastid comprising the steps of introducing into plastids a recombinant nucleic acid molecule or vector, and two phases of selection with first selection phase using a non-lethal compound and a second selection phase using a lethal compound. Alternatively, the dual selection method is conducted simultaneously using a lower concentration of the lethal compound. |
PLASTID GENETIC ENGINEERING VIA SOMATIC EMBRYOGENESIS, H Daniell (2004). University of Central Florida, USA. Patent # CA2491690.
The instant invention is drawn to a method transforming plastids of non-green plant cells, plastid transformation vectors used in the methods, and plant cells and plants thereby obtained. |
PROCESSES AND VECTORS FOR PLASTID TRANSFORMATION, S Herz (2004). Icon Genetics AG, Germany. Patent # MXPA03006193.
This invention discloses a novel process and vector therefore for producing a multicellular plant or plant cells having stably transformed plastids. This process comprises transforming plastids or plant cells via homologous recombination with a DNA molecule enabling DNA modification, whereby said DNA molecule comprises a fragment of a gene requiring for expression a sequence element of the host plastid. The invention also includes the use of novel selection inhibitors for plastic transformation and the possibility of performing multiple rounds of transformation without accumulation of resistance genes. |
MULTIPLE GENE EXPRESSION FOR ENGINEERING NOVEL PATHWAYS AND HYPEREXPRESSION OF FOREIGN PROTEINS IN PLANTS, H Daniell & W Moar (2004). Universities of Auburn & Central Florida, USA. Patent # US2004210966.
Introducing blocks of foreign genes in a single operon would avoid complications such as position effect and gene silencing inherent in putting one gene at a time into random locations in the nuclear genome. Cloning several genes into a single T-DNA does not avoid the compounded variable expression problem encountered in nuclear transgenic plants. This disclosure shows that a bacterial operon can be expressed in a single integration event as opposed to multiple events requiring several years to accomplish. Expression of multiple genes via a single transformation event opens the possibility of expressing foreign pathways or pharmaceutical proteins involving multiple genes. Expressing the Cry2aA2 operon, including a putative chaperonin to aid in protein folding, in the chloroplast via a single transformation event leads to production of crystalised insecticidal proteins. Expressing the Mer operon via a single transformation event leads to a phytoremediation system. |
UNIVERSAL VECTOR FOR STABLE TRANSFORMATION OF CHLOROPLAST GENOME, METHOD FOR STABLE TRANSFORMATION OF PLANT-TARGET, METHOD FOR CONFERRING TO PLANT RESISTANCE TO PLANT-TARGET AGAINST HERBICIDE, METHOD FOR ASSAY OF TRANSFORMATION OF CHLOROPLAST AND STABLY TRANSFORMED CHLOROPLAST GENOME, H Daniell (2004). University Auburn, USA. Patent # RU2238324.
Vector involves the expression cassette comprising a sequence encoding the interesting peptide that is joined with regulatory sequences providing expressing of the encoding sequence in chloroplasts of plant-target. Cassette-flanking sequences are homologous with conservative spacer sequences of chloroplast genome of target that provides the stable integration of encoding sequence into chloroplast genome. Insertion of mentioned vector into genome of plants of different species allows conferring to them new properties that are stably inherited in progeny. |
METHOD FOR RECOMBINATING PLASTID USING PROCARYOTIC RECOMBINASE GENE, J-R Liu, W-J Jeong, S-R Min, S-W Jeong & S-K Han (2003). Korea Research Institute of Bioscience, S Korea. Patent # WO03060137.
The objective of this invention is to enhance the efficiency of plastid transformation using nuclear transformed plants in which the microbial recombinase A (recA) is targetted to (or expressed in) the plastid. This invention will be better explained by the following detailed descriptions. A plant is transformed with a nuclear transformation vector containing the microbial recA gene added with a plastid targeting sequence. In this nuclear transformed plant, the frequency of plastid transformation is enhanced greater than two-folds due to increased homologous recombination between the plastid transformation vector carrying genes of interest (or target genes) and the plastid genome. In addition, because plastid transformation is accomplished through a gradual process, adventitious shoots selected after being subjected to plastid transformation should be cut into explants, and then shoots regenerated from the explants are to be reselected until all of the plastids in the shoots are uniformly transformed. However, when the nuclear transformed plant is used, the number of reselection is reduced to 1/2 to 1/3 due to increased homologous recombination. |
NUCLEIC ACID SEQUENCES CAPABLE OF IMPROVING HOMOLOGOUS RECOMBINATION IN PLANTS AND PLANT PLASTIDS, L A Gilbertson & J M Staub (2003). Monsanto Co., USA. Patent # US2003113921.
A method for improved plastid transformation efficiency via homologous recombination and nucleic acid sequences useful therefore is provided. Nucleic acid sequences comprising a 5 base pair recombination sequence motif or multiple direct repeats thereof that increase the frequency of integration of a selected transgene through plastid transformation by homologous recombination are provided. |
FUSION PROTEIN EXPRESSION IN PLASTIDS, V S Reddy & S Leelavathi (2002). International Centre for Genetic Engineering & Biotechnology, Italy. Patent # GB2364705.
A method of producing a protein of interest by expression of a polynucleotide encoding a fusion protein in a plastid. The protein of interest is linked to a fusion protein partner wherein the fusion protein has a greater half-life than the individually expressed protein of interest, is resistant to internal cleavage in vivo and comprises a cleavage site, preferably IEGR. Methods of protein purification, plastid transformation to create transplastomic plastids and plant propagation are also claimed. Also claimed is the use of a polynucleotide encoding a fusion partner to increase the stability of a recombinantly expressed protein of interest in a plastid. |
TRANSGENIC PLANTS EXPRESSING MinD OR MinE AND AN EFFICIENT METHOD FOR PLANT CHLOROPLAST TRANSFORMATION AND GENE EXPRESSION, R Dinkins, M S S Reddy & G B Collins (2002). Kentucky, USA. Patent # US2002144309.
The present invention concerns transgenic plant which contain large chloroplasts. The transgenic plant of the present invention comprises within its genome a foreign MinD or MinE gene or a foreign gene which expresses a protein which has the same functional activity as the Arabidopsis thaliana MinD or MinE protein. The present invention further concerns a method of producing the transgenic plants of the present invention which contain large chloroplasts. Finally, the present invention concerns a method of transforming the chloroplasts genome of the transgenic plants of the present invention which contain large chloroplasts. |
PLASTID TRANSFORMATION, V S Reddy, L Sadhu, V Shukla & G Ferraiolo (2001). International Centre for Genetic Engineering & Biotechnology, Italy. Patent # WO0142441.
The present invention relates to a method of obtaining a stable transplastome, which method comprises transforming a recipient plastome with a polynucleotide comprising: (a) a 5' sequence homologous to a region of the recipient plastome, and, joined thereto; (b) a sequence heterologous to the recipient plastome comprising a coding region operably linked to at least one regulatory region capable of securing expression of the coding region in the plastid, and, joined thereto; (c) a 3' sequence homologous to a region of the recipient plastome. |
METHOD FOR THE TRANSFORMATION OF PLANT CELL PLASTIDS, J M Staub, G Ye & D L Broyles (2001). Monsanto Co., USA. Patent # WO0181605.
Novel compositions and methods useful for genetic engineering of plant cells to provide a method of producing plastid transformed plants are provided in the instant invention. In particular, the present invention provides methods for obtaining plastid transformed plants on medium containing plastid lethal compounds. |
SHUFFLING OF AGROBACTERIUM AND VIRAL GENES, PLASMIDS AND GENOMES FOR IMPROVED PLANT TRANSFORMATION, L A Castle, M Lassner, K McBride & J English (2001). Maxygen Inc., USA. Patent # WO0138513.
Methods for evolving plant vectors with improved characteristics by recursive recombination are provided. Plant vectors that are RNA or DNA polynucleotides, conjugated-DNA polynucleotides, and plasmids are provided, as are vectors that are Agrobacterium strains and plant viruses. Agrobacterium vectors that have evolved such desired properties as broad host range, increased transformation efficiency, insert precision, targeted insertion, and targeting of T-DNA sequences to the chloroplast are provided. Agrobacterium strains, which are amenable to transforming a broad range of host species using simple transformation techniques such as vacuum infiltration or direct infection in planta, are provided. Plant virus vectors are provided that have evolved desired properties, including: rapid systemic spread, reduction of symptoms, and increased protein expression. Use of the evolved vectors to produce transgenic plants is provided. Methods and vectors for producing proteins in transgenic plants and for conferring pathogen-derived resistance are provided. |
MARKER FREE TRANSGENIC PLANTS: ENGINEERING THE CHLOROPLAST GENOME WITHOUT THE USE OF ANTIBIOTIC SELECTION, H Daniell (2001). Universities of Auburn & Central Florida, USA. Patent # WO0164023.
The present invention provides for a method to circumvent the problem of using antibiotic resistant selectable markers. In particular, target plants are transformed using a plastid vector which contains heterologous DNA sequences coding for a phytotoxin detoxifying enzyme or protein. The selection process involves converting a antibiotic-free phytotoxic agent by the expressed phytotoxin detoxifying enzyme or protein to yield a nontoxic compound. The invention provides for various methods to use antibiotic-free selection in chloroplast transformation. |
METHODS FOR TRANSFORMING PLASTIDS, S Chaudhuri (2000). Calgene LLC, USA. Patent # WO0032799.
An improved method is provided for the transformation of a plant cell plastid. The improved method allows for the increased efficiency of the foreign DNA penetrating the plastid membrane. The method generally involves the use of a plant tissue source having an altered plastid morphology in plastid transformation methods. The present invention finds use in plastid transformation methods for a wide variety of plant species. |
METHOD OF TRANSFORMING PLANT AND TRANSFORMED PLANT, H Nakashita, I Yamaguchi, K Yoshioka & Y Doi (2000). Rikagaku Kenkyusho, Japan. Patent # JP2001046073.
To efficiently carry out the transformation of plants capable of expressing a polycistronic gene by connecting a promoter and an operon containing plural objective genes to a vector and integrating the resultant recombinant vector into a plastid chromosome. The transformation of plants selected from the group consisting of the families Solanaceae, Gramineae, Malvaceae, Brassicaceae, Compositae, Pedaliaceae, Oleaceae, Myrtaceae. Rosaceae, Camelliaceae, Leguminosae, Palmae, Sterculiaceae and Rubiaceae is carried out by connecting a promoter and an operon containing 2-100 kinds of objective genes to a vector and integrating the resultant recombinant vector into a plastid chromosome. Further, it is favorable that the plant belonging to the family Solanaceae is Nicotiana tabacum L. and three kinds of objective genes are polyester synthase gene, & beta -ketothiolase gene and acetoacetyl Co A reductase gene. |
DNA CONSTRUCTS AND METHODS FOR STABLY TRANSFORMING PLASTIDS OF MULTICELLULAR PLANTS AND EXPRESSING RECOMBINANT PROTEINS THEREIN, P Maliga, Z S Maliga, J M Staub, O Zoubenko, L A Allison, H Carrer & I Kanevski (1999). Rutgers, The State University of New Jersey, USA. Patent # US5877402.
DNA constructs are provided for stable transformation of plastids of multicellular plants and expression of foreign proteins in plastids. The DNA constructs comprise a transforming DNA which is targeted to a pre-determined location on the plastid genome and inserted into the plastid genome by homologous recombination with targeting segments comprising DNA sequences homologous to the pre-determined region of the plastid genome. The transforming DNA contains a non-lethal selectable marker gene which confers a selectable phenotype on cells having plastids in which substantially all of the genomes therein contain the transforming DNA (i.e., homoplasmic cells or tissues). The transforming DNA further comprises at least one insertion site for an additional DNA segment, such as a gene encoding a protein for improving a characteristic of the transformed plant. The non-lethal selectable marker gene is preferably provided as a chimaeric gene by assembly from an expression cassette comprising 5' and 3' regulatory segments, preferably derived from plastid genes. A coding segment encoding the non-lethal selectable marker is inserted between the 5' and 3' regulatory segments to form the chimaeric gene. The non-lethal selectable marker coding segment preferred in the present invention is the coding region of aadA from bacteria, which encodes aminoglycoside 3''-adenylyltransferase to confer spectinomycin and streptomycin resistance. |
UNIVERSAL CHLOROPLAST INTEGRATION AND EXPRESSION VECTORS, TRANSFORMED PLANTS AND PRODUCTS THEREOF, H Daniell (1999). Auburn University, USA. Patent # WO9910513.
The invention provides universal chloroplast integration and expression vectors which are competent to stably transform and integrate genes of interest into chloroplast genome of multiple species of plants. Transformed plants and their progeny are provided. Monocotyledonous and dicotyledonous plants are transformed which have never been transformed heretofore. Plants transformed with a synthetic gene express valuable biodegradable protein-based polymers (PBPs). Transformed plants produce high value molecules. Resistance is provided to agricultural crops against the major classes of chemical herbicides. Herbicide resistance is used as a lethal selectable marker for chloroplast transformation. The transformed plants are capable of expressing in addition to the targeted trait, a desirable, secondary non-targeted trait. Insect resistance is provided to transformed plants, both against insects that are susceptible to Bt toxins and against insects that have developed resistance to Bt toxins.
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GENETIC ENGINEERING OF PLANT CHLOROPLASTS, H Daniell & B A McFadden (1999). University of Auburn, USA. Patent # US5932479.
Methods for the transformation of etioplasts and of chloroplasts are disclosed. One method comprises incubating isolated etioplasts with a chelating agent to bind metal ions on which chloroplast nucleases are dependent and incubating the resultant nuclease-inactivated etioplasts with foreign DNA. By an alternative method of transforming chloroplasts, foreign DNA comprising expression cassettes are coated on metal particles and inserted by high-velocity impact into plant cells. For transformation into a plant chloroplast, DNA molecules containing an expression cassette includes a DNA fragment containing appropriate control sequences. The expression cassette may also be flanked by chloroplast DNA which facilitates stable integration of the gene(s) of interest into the recipient chloroplast genome. It is desirable to include within the expression cassette a selectable marker gene that encodes a selectable phenotype which allows for the identification of the cells expressing the introduced gene. Transformed chloroplasts, transformed etioplasts, transformed cells and their progeny, and plants are disclosed. Other aspects of the invention are described.
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PLASTID PROMOTERS FOR TRANSGENE EXPRESSION IN THE PLASTIDS OF HIGHER PLANTS, P Maliga, D Silhavy & P Sriraman Priya (1998). Rutgers University, USA. Patent # WO9855595.
The present invention provides promoter elements useful for stably transforming the plastids of higher plants. The constructs described herein contain unique promoters that are transcribed by both nuclear encoded plastid RNA polymerases, plastid encoded plastid RNA polymerases or both. Use of the novel constructs of the invention facilitates transformation of a wider range of plant species and enables ubiquitous expression of a transforming DNA in plastids of multicellular plants. |
NUCLEAR-ENCODED TRANSCRIPTION SYSTEM IN PLASTIDS OF HIGHER PLANTS, P Maliga, L A Allison & P T Hajdukiewicz (1997), Rutgers University, USA. Patent # WO9706250.
The present invention provides novel DNA constructs and methods for stably transforming the plastids of higher plants. The constructs described herein contain unique promoters that are transcribed by both nuclear encoded plastid polymerases and plastid encoded plastid polymerases. Use of the novel constructs of the invention facilitates transformation of a wider range of plant species and enables tissue specific expression of a transforming DNA in plastids of multicellular plants. |
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