Abstract: A process of producing a transgenic multi-cellular plants or parts thereof expressing a trait of interest, said trait having a controlled distribution of said trait to progeny, wherein said process comprises (i) producing a first plant or a cell thereof having in a first locus of a nuclear chromosome a first heterologous nucleotide sequence comprising a first fragment of a nucleotide sequence encoding said trait of interest, (ii) producing a second plant or a cell thereof having in a second locus of a nuclear chromosome homologous to said nuclear chromosome of step (i), a second heterologous nucleotide sequence comprising a second fragment of the nucleotide sequence encoding said trait of interest, and (iii) hybridising said first and said second plant or cells thereof to generate progeny exhibiting said functional trait of interest due to binding between a protein or polypeptide encoded by said first heterologous nucleotide sequence and a protein or polypeptide encoded by said second heterologous nucleotide

Abstract: The invention relates to a novel circuit arrangement for electrotransfection or electrofusion, which enables the transportation of DNA and/or other biologically active molecules to the nucleus of higher eukaryotic cells or the fusion of cells, independent of cell division and with reduced cell mortality.

Abstract: Exemplary methods include a method for transforming an algal cell by preparing a transformation construct, preparing a particle for bombarding the algal cell, adhering the transformation construct to the particle, bombarding the algal cell with the particle, and growing the algal cell into a colony. The transformation construct is replicated within a nuclear genome of the algal cell and the growing of the algal cell is in a nutrient medium. Another exemplary method may include a method for genetically modifying an algal cell, by adding nucleic acid to the algal cell while the algal cell is suspended in a solution of low conductivity, introducing the nucleic acid into the algal cell by application of an electrical pulse resulting in a transformed algal cell, and selecting a colony that includes the transformed algal cell.

Abstract: Methods for introducing a functionalized linear nucleic acid cassette molecule of interest into a plant cell comprising a cell wall include use of nanoparticles. In some embodiments, the cell comprising a cell wall is a cultured plant cell. Methods include genetically or otherwise modifying plant cells and for treating or preventing disease in any plant, especially crop plants. Transgenic plants include a nucleic acid molecule of interest produced by regeneration of whole plants from plant cells transformed with functionalized linear nucleic acid cassette molecules.

Abstract: Methods for introducing a linear nucleic acid molecule of interest into a cell comprising a cell wall include use of nanoparticles coated with polyethylene glycol. In some embodiments, the cell comprising a cell wall is a plant cell. Methods include genetically or otherwise modifying plants and for treating or preventing disease in plant cells comprising a cell wall. Transgenic plants include a nucleic acid molecule of interest produced by regeneration of whole plants from plant cells transformed with linear nucleic acid molecules.

Abstract: Methods for the transformation of sugar cane are provided. The methods comprise utilizing sugar cane immature shoots as the source of plant material for transformation. Segments of the immature shoot are excised and transformed by any suitable transformation methodology. In some embodiments, the segments are cultured in embryogenic culture induction medium prior to transformation. Transformation can be performed via Agrobacterium-mediated gene delivery, biolistic transformation, and the like. Transgenic plants are regenerated from plantlets grown under conditions favoring growth of transformed cells while substantially inhibiting growth of non-transformed cells.

Abstract: According to the present invention, a technique of increasing the frequency of genetic recombination in genomic DNA of a plant is established. Such technique comprises: introducing a restriction enzyme gene that can be expressed in a target plant cell into the plant cell and causing the restriction enzyme gene to be transiently expressed so as to induce genetic recombination of genomic DNA; or introducing a promoter and a restriction enzyme gene using the Agrobacterium method so as to induce genetic recombination of genomic DNA.

Abstract: An electroporation cuvette is constructed with electroporation electrodes arranged in non-parallel relation to form a gap whose width varies with the location within the cuvette, plus a pair of positioning electrodes that are arranged to cause electrophoretic migration of biological cells within the cuvette according to cell size. Once the cells, suspended in a solution of the impregnant, are distributed in the cuvette by the positioning electrodes, electric field pulses are generated by the non-parallel electroporation electrodes. Because of their distribution in the cuvette, the various cells will experience voltage differentials across their widths that approach uniformity regardless of cell diameter, since the larger cells will be positioned at locations where the gap between the electrodes is greater and the smaller cells at locations where the gap is relatively small while the voltage drop across the entire gap is uniform along the length of the cell.

Abstract: The present invention provides a method for selecting genomic DNA fragments which are useful for providing a plant with an agriculturally advantageous improvement. The method of the present invention comprises the steps of: 1) preparing genomic DNA from a plant, which is then cloned into a cloning vector to form a genomic DNA library; 2) introducing a genomic fragment from each of the genomic clones constituting the genomic DNA library separately into a plant to produce transgenic plants; 3) cultivating the transgenic plants or progeny thereof to select a plant exhibiting an agriculturally advantageous phenotypic variation; and 4) selecting the genomic DNA fragment, which was introduced in step (2) into the plant selected in step (3), as a purposed genomic DNA fragment.

Abstract: The introduction of genetic material or molecules of biological interest into cells is a procedure with an increasing interest both for experimental and application purposes, so that electroporation is a widely used technique, but the electroporation of single adhering cells is still impaired. The present application describes an apparatus for the electroporation of any kind of cell adhering to a substrate at any stage of development, where an electrical signal can be driven and applied to a single adhering cell in culture in order to obtain its electroporation. The method to electroporate a single adhering cell with the apparatus of the invention is also described.

Abstract: A method for producing transgenic plants, including treating a target tissue using plasmolyzing media (PM) which contains 4% to 10% of sucrose and 100 ?M to 300 ?M of Acetosyringone (AS) and gold particles. The target tissue is infected by a bacterial suspension using a suitable strain and a suitable transformation vector. A PM containing 4% to 10% sucrose and 100 ?M to 300 ?M AS is treated for a period between 1 to 3 days. Cultivation is performed in a cultivation media in a dark condition at a temperature between 25° C. to 30° C. A non-selection media with an antibiotic is introduced. A selection media containing an active ingredient phosphinothricin (PPT) is introduced in a light condition at a temperature of between 25° C. to 30° C. in a sub culture for a period of between 3 weeks to 1 month. The putative transformant is regenerated and the number of copies of the transgenes is analyzed.

Abstract: A system and method are described for electroporating a sample that utilizes one or more sets of electrodes that are spaced apart in order to hold a surface tension constrained sample between the electrodes. The first electrode is connected to the lower body of the system while the second electrode is connected to the upper body. Both electrodes are connected to a pulse generator. Each electrode has a sample contact surface such that the first electrode and the second electrode may be positioned to hold a surface tension constrained sample between the two sample contact surfaces and the sample may receive a selected electric pulse.

Abstract: There is provided a method by which multiple types of substances desired to be transferred into cells can be continuously transferred into multiple types of cells by a convenient procedure, a cell in which the substance desired to be transferred into cells has been taken up by this method, and an apparatus for transferring a substance into cells by this method. The foregoing objects can be achieved by electrospraying cells with a liquid free from the substance to be transferred into cells while the cells are kept in contact with the substance to be transferred into cells, or first electrospraying cells with a liquid free from the substance to be transferred into cells and then bringing the cells into contact with the substance to be transferred into cells.

Abstract: The present invention provides a method and apparatus for transferring an agent into a cell. The method includes the steps of providing an agent outside of a cell and generating a vapor bubble and a plasma discharge between an avalanche electrode and a conductive fluid surrounding the cell. The vapor bubble and plasma discharge generate a mechanical stress wave and an electric field, respectively. The combination of this mechanical stress wave and electric field results in permeabilization of the cell, which in turn results in transfer of the agent into the cell.

Abstract: A microinjection apparatus has a trap plate which traps at least one cell, so as to fix a position thereof, a capillary needle which injects a substance into the cell trapped by the trap plate, a movement mechanism portion which thrusts a distal end of the capillary needle onto a in a lengthwise direction of the needle, and which withdraws the capillary needle in the lengthwise direction thereof, and a discharge control portion which discharges the substance from the distal end of the capillary needle, when the capillary needle has been withdrawn to a predetermined position by the movement mechanism portion.

Abstract: The capillary gun for delivery of ballistic particles to a target includes an inner capillary tube disposed concentrically within an outer capillary tube with the input end of the inner tube connected to a channel through which a continuous flow of high speed helium gas carrying ballistic particles is introduced. The outer capillary tube, which is connected to a vacuum source, has an outlet end that extends slightly beyond the end of the inner tube. A cap placed over the output end of the outer tube has an opening at its center through which the particles exit the device. The vacuum source applies continuous suction to the space between the outer tube and the inner tube, drawing the gas from the output end of the inner tube while the inertia of the accelerated particles causes them to continue in the axial direction through the exit opening for delivery to the target. Multiple particle injectors provide for the concurrent injection of different materials without disruption of the gas flow.

Abstract: Methods for transforming an explant are provided. The methods may include applying copper amino acid chelate to a plant and transforming an explant obtained therefrom. The transformed explant may have increased transformation frequency relative to a control.

Abstract: The present invention relates to a method of introducing nucleic acids into cells by electroporation, comprising the step (A) of loading nucleic acids to the surface of an electrode; the step (B) of adhering cells on the obtained nucleic acid-loaded electrode surface; and the step (C) of applying electric pulses to the adhering cells. According to this method, not only efficient introduction of a gene into cells but also gene introduction at desirable timing and at desirable sites can be performed without damaging the adhering cells.

Abstract: The present disclosure provides methods, recombinant DNA molecules, recombinant host cells containing the DNA molecules, and transgenic and genetically engineered plant cells, plant tissue, seeds and plants which contain and express an herbicide resistant protoporphyrinogen oxidase such that they germinate from seed and grow in the presence of an amount of herbicide where the parent plant does not. Such plants are especially appropriate for use in agriculture or horticulture where herbicides are used to kill undesirable plants which might contaminate or compete with the transgenic plant of interest.

Abstract: GmWRKY transcription factor genes and proteins from soybean. Plants that overexpress GmWRKY transcription factor genes and proteins to thereby increase stress tolerance. Methods for making such slants and methods for mimicking a stress tolerance phenotype using a GmWRKY modulator.

Abstract: Methods for treating or preventing infections from coagulase-negative staphylococci using proteins and polypeptides from coagulase-negative staphylococcal bacteria such as S. epidermidis, including proteins designated SdrF, SdrG and SdrH, and their effective fragments such as their respective A domains, are provided. Methods are also provided wherein antibodies that recognize the SdrG protein or its ligand binding A region are used to treat or prevent staphylococcal infection, and these methods can also be utilized to prevent the formation of infections on indwelling medical devices.

Abstract: Methods and means are provided to improve targeted DNA insertion in plants using rare-cleaving “double stranded break” including enzymes. Also provided are improved I-SceI encoding nucleotide sequences.

Abstract: A process of producing a transgenic multi-cellular plants or parts thereof expressing a trait of interest, said trait having a controlled distribution of said trait to progeny, wherein said process comprises (i) producing a first plant or a cell thereof having in a first locus of a nuclear chromosome a first heterologous nucleotide sequence comprising a first fragment of a nucleotide sequence encoding said trait of interest, (ii) producing a second plant or a cell thereof having in a second locus of a nuclear chromosome homologous to said nuclear chromosome of step (i), a second heterologous nucleotide sequence comprising a second fragment of the nucleotide sequence encoding said trait of interest, and (iii) hybridising said first and said second plant or cells thereof to generate progeny exhibiting said functional trait of interest due to binding between a protein or polypeptide encoded by said first heterologous nucleotide sequence and a protein or polypeptide encoded by said second heterologous nucleotide

Abstract: The present invention relates to a gene gun and the application of the gene gun for gene transformation. A low pressure gas is used in the gene gun to directly accelerate the biological material containing solution, so that the biological materials penetrate through the cell membrane/wall or the skin of an animal, without using metal particle carriers, for gene transformation.

Abstract: The present invention is directed to the introduction of molecules, including nucleic acids, carbohydrates, plant growth regulators and peptides into cells and tissues. The present invention is also directed to media and methods for enhancing embryogenic callus production of elite lines of soybean.

Abstract: A process of producing a transgenic multi-cellular plant or animal organism expressing a trait of interest and having a controlled distribution of said trait to progeny, wherein said process comprises hybridising a first multi-cellular organism or a cell thereof having a first heterologous DNA sequence comprising a first fragment of a nucleotide sequence encoding said trait of interest and a second multi-cellular organism or a cell thereof having a second heterologous DNA sequence comprising a second fragment of the nucleotide sequence encoding said trait of interest, whereby said first and said second heterologous sequences are designed such that said trait of interest arises due to RNA trans-splicing after said hybridation.

Abstract: This invention relates to an isolated nucleic acid fragment encoding a sucrose transport protein. The invention also relates to the construction of a chimeric gene encoding all or a portion of the sucrose transport protein, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the sucrose transport protein in a transformed host cell.

Abstract: The present invention involves methods and devices which enable discrete objects having a conducting inner core, surrounded by a dielectric membrane to be selectively inactivated by electric fields via irreversible breakdown of their dielectric membrane. One important application of the invention is in the selection, purification, and/or purging of desired or undesired biological cells from cell suspensions. According to the invention, electric fields can be utilized to selectively inactivate and render non-viable particular subpopulations of cells in a suspension, while not adversely affecting other desired subpopulations. According to the inventive methods, the cells can be selected on the basis of intrinsic or induced differences in a characteristic electroporation threshold, which can depend, for example, on a difference in cell size and/or critical dielectric membrane breakdown voltage.

Abstract: Various embodiments provide, for example, vectors, expression cassettes, and cells useful for transgenic expression of nucleic acid sequences. In various embodiments, vectors can contain plastid-based sequences of unicellular photosynthetic bioprocess organisms for the production of food- and feed-stuffs, oils, biofuels, pharmaceuticals or fine chemicals.

Abstract: Compositions and methods for protecting a plant from an insect pest are provided. In particular, nucleic acid sequences encoding insect protoxins modified to comprise at least one proteolytic activation site that is sensitive to a plant protease or an insect gut protease are provided. Cleavage of the modified protoxin at the proteolytic activation site by a protease produces an active insect toxin. Methods of using the modified insect protoxin nucleic acid sequences and the polypeptides they encode to protect a plant from an insect pest are provided. Particular embodiments of the invention further provide modified insect protoxin compositions and formulations, expression cassettes, and transformed plants, plant cells, and seeds.

Abstract: Provided are methods of increasing the tolerance of a plant to abiotic stresses and/or increasing the biomass and/or increasing the yield of a plant by expressing within the plant an exogenous polynucleotide homologous to SEQ ID NO:13.

Abstract: The electroporation array is comprised of three technologies: microwire glass electrodes, microelectronic multiplexer stimulator chips and microfluidic flow chamber. Various substances, such as genes, gene silencing RNAi, gene inhibition agents or drugs, can be perfused into the microfluidic flow chamber. The entry of the various substances into the cells will be facilitated by electroporation. An applied electric potential causes nanoscale pores to open in the cell membrane allowing substances in the solution to freely diffuse into the cell. The specific cells selected for electroporation are defined using the computer controlled microelectronic stimulator array. An “image” of which electrodes within the array to apply the electric potential to, and thus electroporate, is de-multiplexed onto the array. All the selected electrodes deliver a current pulse varied by the intensity of the de-multiplexed “image”.

Abstract: Chamber for treating cells contained in a suspension in the electrical field with a beaker made from electrically non-conductive material, into which an elongate core made from electrically non-conductive material is at least partially inserted axially through an opening, between the beaker and the core a gap being present to receive the suspension; at least two electrodes made from electrically conductive material arranged on the outer face of the core facing the gap, between which an electrical field can be created to treat cells in a suspension contained in the gap by applying a voltage; the thermal expansion coefficient of the material of the electrodes and of the material of the core being matched with one another such that the electrodes do not substantially alter their position relative to the core in the temperature range from ambient temperature to temperatures reached during autoclaving and/or sterilising.

Abstract: The present invention refers to gene expression cassettes encoding enzymes involved in the metabolic pathway for the biosynthesis of hemicelluloses, cellulose and/or uronic acids, and to a method for genetic transformation of plant cells through the introduction of one or more gene expression cassettes from the present invention, and the overexpression and repression of these genes in plants. More particularly, the present invention refers to a method for introducing expression cassettes in plants. Additionally, the present invention refers to genetically modified plants. The present invention also refers to the attainment of the genetically modified plant, its derived plants and seeds, as well as the wood, paper and cellulose from this plant.

Abstract: Described is a method for method for transferring material through the membrane of at least one cell, wherein the transfer is carried out in the presence of trehalose. This method is applicable in particular in the field of genetic engineering and biotechnology.

Abstract: Methods for the targeted integration of nucleotide sequences into a plant are provided. Transfer cassettes comprising nucleotide sequences of interest flanked by non-identical recombination sites are used to transform a plant comprising a target site. The target site contains at least a set of non-identical recombination sites corresponding to those on the transfer cassette. Exchange of the nucleotide sequences flanked by the recombination sites is effected by a recombinase.

Abstract: An oocyte recording chamber for electrophysiological measurements. The recording chamber includes a base and a cover attached to the base. The cover and the base define a chamber having a size sufficient to accommodate an oocyte. The recording chamber includes a first electrode and a second electrode that are positioned so that the tips of the electrodes penetrate the membrane of the oocyte when the cover is fastened to the base. The recording chamber also includes a third electrode and a fourth electrode exposed to the chamber and used as ground electrodes.

Abstract: A site specific recombination system and methods of use thereof are disclosed for manipulating the genome of higher plants.

Abstract: Transgenic plants that express CIVPS or intein modified therapeutic proteins, compositions of matter comprising them, therapeutic proteins made from the transgenic plants, methods to construct the transgenic plants containing CIVPS or intein modified therapeutic genes, methods to express CIVPS or intein modified therapeutic proteins in plants, and methods of using the transgenic plants.

Abstract: The present disclosure provides methods for obtaining the targeted integration of a DNA molecule into the genome of a host cell using a recombinase. The methods disclosed herein can be used with a variety of host cells, including, for example, dicotyledonous and monocotyledonous plant cells. The present disclosure provides a method for effecting site-specific recombination of DNA within a plant cell, comprising: introducing into the plant cell a target nucleotide sequence comprising a first Int recognition site; introducing into the plant cell a donor nucleotide sequence comprising a second Int recognition site; and introducing into the plant cell an integrase or integrase complex.

Abstract: This invention is predicated on the present applicants' discovery that nanostructures comprising discrete regions of different composition can be used to deliver to a biological cell a desired combination of molecules in close proximity. Different molecules can be selectively bonded to discrete regions of different composition in sufficiently close physical relationship to enhance delivery or effectiveness within the cell. The preferred nanostructures are multicomponent nanorods. Important applications include delivery of missing DNA sequences for gene therapy and delivery of antigens or DNA encoding antigens for vaccination.

Abstract: A molecule introducing apparatus is provided with a power supply circuit 20 for supplying power, a step-up transformer 30 which is supplied with power from the power supply circuit 20 and which outputs a high voltage, a switching transistor 22 that blocks off and allows the supply of power from the power supply circuit 20 to the step-up transformer 30 and which generates an instantaneous high voltage, and a pair of electrode probes 50 for applying the instantaneous high voltage generated at the step-up transformer 30 to a predetermined region in a body. The molecule introducing apparatus oscillates DNA arranged outside a cell by the instantaneous high voltage so as to introduce the DNA into the cell.

Abstract: Porous and/or polycrystalline silicon are used in the delivery of substances into cells. The porous and/or polycrystalline silicon can be formed into micropiercers, microneedles and biolistic bullets for penetration of the cell. The control of the pore size and porosity of the porous and/or polycrystalline silicon allows tuning of the bioactivity of the porous silicon. The porous and/or polycrystalline silicon is also resorbable and is therefore resorbed from the cells without leaving any particles or being seen as a foreign body. Methods of manufacturing the porous silicon micropiercers, microneedles, microelectrodes, biolistic bullets, and precipitation of calcium phosphate on a bioactive substrate, and their advantages over known methods of delivering materials into cells.

Abstract: An L-amino acid-producing strain of Escherichia coli is bred by modifying an Escherichia coli K12 strain or a derivative thereof so as to become resistant to L-valine and have an ability to produce one or more L-amino acids selected from the group consisting of L-tryptophan, L-phenylalanine, L-lysine, L-tyrosine, L-glutamic acid, L-histidine, L-cysteine, and L-proline.

Abstract: Mutant strains of Rhodococcus equi are disclosed, and vaccines comprising same.

Abstract: A method for producing a transgenic plant, which comprises: (A) introducing a vector into a plant cell, wherein the vector is a vector for gene introduction into a plant and comprises: a desired gene, and a selectable marker gene comprising a gene encoding an enzyme which synthesizes auxin from an auxin precursor; (B) culturing the plant cell into which the genes are introduced by the vector, in the presence of an auxin precursor and/or an analogue thereof to thereby prepare a redifferentiated tissue, and detecting and selecting the redifferentiated tissues; and (C) culturing the redifferentiated tissue selected in (B) to redifferentiate a plant individual, and a vector for gene introduction into a plant, which comprises: a desired gene, and a selectable marker gene comprising an indoleacetamide hydrolase, iaaH, gene and an isopentenyl transferase, ipt, gene and being free of an tryptophan monooxygenase, iaaM, gene.

Abstract: The subject invention pertains to materials and methods for producing plants that are resistant to infection by geminiviruses and other related viruses. Methods of the invention comprise transforming a plant with a polynucleotide wherein when the polynucleotide is expressed in the plant, the transformed plant exhibits resistance to plant viral infections. Exemplified herein is the use of a polynucleotide encoding a Rep protein derived from tomato mottle geminivirus. The methods of the invention can be used to provide resistance to viral infection in plants such as tomato and tobacco. The present invention also concerns transformed and transgenic plants in plant tissue that express a polynucleotide encoding a plant virus Rep protein, or a fragment or variant thereof.

Abstract: This invention relates to an isolated nucleic acid fragment encoding a sucrose transport protein. The invention also relates to the construction of a chimeric gene encoding all or a portion of the sucrose transport protein, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the sucrose transport protein in a transformed host cell.

Abstract: An electrospraying apparatus and/or method is used to coat particles. For example, a flow including at least one liquid suspension may be provided through at least one opening at a spray dispenser end. The flow includes at least particles and a coating material. A spray of microdroplets suspending at least the particles is established forward of the spray dispenser end by creating a nonuniform electrical field between the spray dispenser end and an electrode electrically isolated therefrom. The particles are coated with at least a portion of the coating material as the microdroplet evaporates. For example, the suspension may include biological material particles.

Abstract: A synthetic polynucleotide encoding human lactoferrin, modified with respect to the natural gene so as to maximize its expression in vegetals, on the basis of the preferential use of the codons is described. Moreover, the vectors containing such sequence, that having regulation elements activated in a controlled way determine its tissue- and stage-specific expression are further described. The vegetal cells and the plants transformed with the afore mentioned vectors, as well as the production processes of functional foods, vegetal milks, and human lactoferrin, utilizing them are also described.