Abstract: A novel hybrid maize variety designated 32H20 and seed, plants and plant parts thereof, produced by crossing Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety 32H20 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 32H20 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed 32H20, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid 32H20. This invention further relates to methods for producing maize lines derived from hybrid maize variety 32H20 and to the maize lines derived by the use of those methods.

Abstract: The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites and/or multiple topoisomerase recognition sites. The methods permit the simultaneous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different.

Abstract: A novel hybrid maize variety designated 37Y12 and seed, plants and plant parts thereof, produced by crossing Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety 37Y12 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 37Y12 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed 37Y12, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid 37Y12. This invention further relates to methods for producing maize lines derived from hybrid maize variety 37Y12 and to the maize lines derived by the use of those methods.

Abstract: A novel hybrid maize variety designated 35F38 and seed, plants and plant parts thereof, produced by crossing Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety 35F38 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 35F38 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed 35F38, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid 35F38. This invention further relates to methods for producing maize lines derived from hybrid maize variety 35F38 and to the maize lines derived by the use of those methods.

Abstract: A novel inbred maize line designated PHCJP and seed, plants and plant parts thereof. Methods for producing a maize plant that comprise crossing inbred maize line PHCJP with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into PHCJP through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. Hybrid maize seed, plant or plant part produced by crossing the inbred line PHCJP or a trait conversion of PHCJP with another maize line. Inbred maize lines derived from inbred maize line PHCJP, methods for producing other inbred maize lines derived from inbred maize line PHCJP and the inbred maize lines and their parts derived by the use of those methods.

Abstract: A novel inbred maize line designated PHCER and seed, plants and plant parts thereof. Methods for producing a maize plant that comprise crossing inbred maize line PHCER with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into PHCER through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. Hybrid maize seed, plant or plant part produced by crossing the inbred line PHCER or a trait conversion of PHCER with another maize line. Inbred maize lines derived from inbred maize line PHCER, methods for producing other inbred maize lines derived from inbred maize line PHCER and the inbred maize lines and their parts derived by the use of those methods.

Abstract: The invention provides novel methods of gene targeting using replication in order to increase the efficiency of targeted genetic modification in an eukaryotic organism. Included are vectors, expression cassettes, and modified cells, plants and seeds.

Abstract: A method to produce a cell expressing an antibody from a genomic sequence of the cell comprising a modified immunoglobulin locus using Cre-mediated site-specific recombination is disclosed. The method involves first transfecting an antibody-producing cell with a homology-targeting vector comprising a lox site and a targeting sequence homologous to a first DNA sequence adjacent to the region of the immunoglobulin loci of the genomic sequence which is to be converted to a modified region, so the first lox site is inserted into the genomic sequence via site-specific homologous recombination. Then the cell is transfected with a lox-targeting vector comprising a second lox site suitable for Cre-mediated recombination with the integrated lox site and a modifying sequence to convert the region of the immunoglobulin loci to the modified region.

Abstract: The present invention describes methods of identifying altered recombinases and compositions thereof, wherein at least one amino acid is different from a parent, wild-type recombinase and the altered recombinase has improved recombination efficiency towards wild-type and/or pseudo att site sequences relative to the parent, wild-type recombinase. The present invention also includes methods of modifying the genomes of cells using the altered recombinases, including methods of site-specifically integrating a polynucleotide sequence of interest in a genome of a eucaryotic cell.

Abstract: In the present invention, viruses, plasmids or both are constructed which contain viral DNA and lox sites positioned such that site-specific recombination between lox sites in separate plasmids results in generation of infectious viral DNA at high-efficiency in cotransfected host cells that have been engineered to express the Cre recombinase. Because of the high-efficiency and specificity of the Cre enzyme, suitably engineered plasmids can be readily recombined to produce infectious virus at high-efficiency in cotransfected 293 cells, without, at the same time, producing wild-type adenovirus, with the attendant problems for removal thereof. Use of recombinases besides Cre and recombinase recognition sites besides lox sites, and use of cells other than 293 cells are also disclosed and enabled, as are kits incorporating the site-specific vector system.

Abstract: The present invention relates generally to transgenic plants. More specifically, it relates to methods and compositions for the introduction of DNA using circular molecules that are not able to replicate outside a host cell. The circular molecules contain site-specific recombination sequences and allow transformation of host cells with DNA comprising only selected sequences of interest.

Abstract: The present invention relates to novel methods for the transformation of crop species, novel methods for the selection and identification of transformed plant cells and novel methods for recovery of regenerated whole plants. The method also relates to the development of plants with novel traits and plants that contain novel recombinant DNA constructs.

Abstract: The invention provides methods of covalently joining nucleic acid molecules and methods of molecular cloning. The methods provide either sequential or simultaneous ligation of flanking or vector nucleic acid molecules to nucleic acid insert molecules by topoisomerase and DNA ligase. The methods provide for directional and non-directional covalent joining and cloning of nucleic acid molecules.

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: A novel hybrid maize variety designated 33Y45 and seed, plants and plant parts thereof, produced by crossing two Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety 33Y45 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 33Y45 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed 33Y45, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid 33Y45. This invention further relates to methods for producing maize lines derived from hybrid maize variety 33Y45 and to the maize lines derived by the use of those methods.

Abstract: A novel hybrid maize variety designated X0923B and seed, plants and plant parts thereof, produced by crossing two Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety X0923B with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into X0923B through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed X0923B, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid X0923B. This invention further relates to methods for producing maize lines derived from hybrid maize variety X0923B and to the maize lines derived by the use of those methods.

Abstract: Tn5 transposase (Tnp) mutants that have higher transposase activities than the wild-type Tnp are disclosed. The Tn5 Tnp mutants differ from the wild-type Tnp at amino acid positions 54, 242, and 372 and have greater avidity than the wild-type Tnp for at least one of a wild-type Tn5 outside end sequence as defined by SEQ ID NO:3 and a modified Tn5 outside end sequence as defined by SEQ ID NO:5. Also disclosed are various systems and methods of using the Tnp mutants for in vitro or in vivo transposition.

Abstract: A novel hybrid maize variety designated 32R38 and seed, plants and plant parts thereof, produced by crossing two Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety 32R38 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 32R38 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed 32R38, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid 32R38. This invention further relates to methods for producing maize lines derived from hybrid maize variety 32R38 and to the maize lines derived by the use of those methods.

Abstract: The present invention describes methods for producing antibody libraries, and particularly for increasing antibody library diversity by inducing mutagenesis within the CDR regions of immunoglobulin heavy or light chains that are displayed on the surface of filamentous phage particles comprising the library. The invention also describes oligonucleotides useful for increasing the library diversity, and universal light chains useful in the library production methods.

Abstract: A novel hybrid maize variety designated 34B39 and seed, plants and plant parts thereof, produced by crossing two Pioneer Hi-Bred International, Inc. proprietary inbred maize lines. Methods for producing a maize plant that comprises crossing hybrid maize variety 34B39 with another maize plant. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 34B39 through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby. This invention relates to the hybrid seed 34B39, the hybrid plant produced from the seed, and variants, mutants, and trivial modifications of hybrid 34B39. This invention further relates to methods for producing maize lines derived from hybrid maize variety 34B39 and to the maize lines derived by the use of those methods.

Abstract: The present invention provides a method comprising a step of transferring a cell proliferation factor gene into a mammalian liver cell to obtain an immortalized liver cell, a step of proliferating the immortalized liver cell, and a step of removing the cell proliferation factor gene from the immortalized liver cell; a large number of liver cells obtained thereby; and a treating agent and a artificial liver comprising obtained liver cell.

Abstract: Adenovirus packaging cell lines for growth of an E1A/E1B deficient adenovirus that is substantially free of replication competent adenovirus (RCA) are provided. Methods for producing adenovirus substantially free of RCA are also provided, wherein the adenovirus is grown in a cell line containing coding sequences for adenovirus E1A and E1B, which are operably linked to promoters that lack polynucleotide sequences sharing substantial sequence identity with the native adenovirus E1A and E1B promoters.

Abstract: A method of transferring a foreign gene into cells, characterized by involving: the step of transferring into the cells with the use of an adenovirus vector, a first nucleic acid, which has a sequence provided with adeno-associated virus-origin ITRs in both sides of the target foreign gene to be transferred, and a second nucleic acid, which has an adeno-associated virus-origin rep gene and a promoter for expressing this gene and carries a stuffer sequence inserted thereinto sandwiched in two recombinase recognition sequences and located between the rep gene and the promoter; and the step of expressing the Rep protein under the action of recombinase in the cells obtained in the above step to thereby integrate the target foreign gene into the chromosomal DNA.

Abstract: The invention claims a class of adenovirus vectors for delivering recombinases to a large number of cells of different origins, and methods for regulating the expression of a gene in transfected mammalian cells in culture and in cells of transgenic animals, comprising infecting said cells with an Ad vector encoding a recombinase whose target site is present at or adjacent to the gene, wherein the action of the recombinase regulates the expression of said gene.

Abstract: Cloning systems are provided for constructing expression vectors. In one aspect of the invention, a kit is provided for constructing one or more recombinant expression vectors. The kit comprises: a linear driver DNA comprising a promoter sequence, a donor recombination site, and at least one selectable marker, the linear driver DNA being capable of being ligated with one or more linear donor DNA comprising a donor DNA sequence to form one or more circular donor DNA; and a circular acceptor vector comprising an origin of replication and an acceptor recombination site capable of recombining with the circular donor DNA to form the recombinant expression vector for expressing the donor DNA sequence.

Abstract: This invention concerns a method for effecting directed mutagenesis based on a guided homologous recombination system that comprises (i) a triple helix forming oligonucleotide, (ii) a donor nucleic acid segment, and (iii) an adapter segment comprising an oligonucleotide sequence able to bind at least a portion of said donor nucleic acid through Watson-Crick base pairing.

Abstract: Methods and vectors and kits are provided for producing chimeric nucleic acid constructs capable of producing dsRNA for silencing target nucleic acid sequences of interest using recombinational cloning.

Abstract: Site-specific recombinase based methods for making a recombinant adenoviral genome, as well as kits for practicing the same and the recombinant adenovirus vectors produced thereby, are provided. In the subject methods, the subject genomes are prepared from donor and acceptor vectors that each include at least one site recombinase recognition site, where in certain preferred embodiments, one of the donor and acceptor vectors includes a single recombinase recognition site while the other includes two recombinase recognition sites. The acceptor vector includes an adenoviral genome having an E region deletion. The donor vector includes an insertion nucleic acid. In the subject methods, the donor and acceptor vectors are combined in the presence of a recombinase to produce an adenoviral genome that includes the insertion nucleic acid. The subject adenoviral genomes find use in a variety of applications, including as vectors for use in a variety of applications.

Abstract: The present invention relates to methods of producing an allelic series of modifications in genes of interest in a cell. In particular, the invention provides methods for using nucleic acid sequence-modifying agents (e.g., chemicals, electromagnetic radiation, etc.) to introduce modifications in any nucleic acid sequence in the genome of a cell. Also provided are sets of cells which contain at least one modification in any gene of interest. The methods and compositions of the invention are useful in determining the function of the gene of interest.

Abstract: The invention provides a novel Adenovirus backbone plasmid, which when co-transfected with a shuttle vector, allows for production of recombinant viruses quickly and easily. The present invention also provides host cells and a cloning system for generating recombinant adenoviruses.

Abstract: A method of screening for a target molecule from a group of potential target molecules is described. This method involves using a library of lentiviral vectors where the members encode the group of target molecules, then transducing a group of cells and screening the transduced cell for a desired phenotype. The cell(s) displaying the desired phenotype is selected and the target molecule is identified.

Abstract: The present invention provides compositions, including vectors, and methods for the rapid subcloning of nucleic acid sequences in vivo and in vitro. In particular, the invention provides vectors used to contain a gene of interest that comprise a sequence-specific recombinase target site. These vectors are used to rapidly transfer the gene or genes of interest into any vector that contains a sequence-specific recombinase target site located downstream of a regulatory element so that the gene of interest may be regulated.

Abstract: A retroviral delivery system capable of transducing a target site is described. The retroviral delivery system comprises a first nucleotide sequence coding for at least a part of an envelope protein; and one or more other nucleotide sequences derivable from a retrovirus that ensure transduction of the target site by the retroviral delivery system; wherein the first nucleotide sequence is heterologous with respect to at least one of the other nucleotide sequences; and wherein the first nucleotide sequence codes for at least a part of a rabies G protein or a mutant, variant, derivative or fragment thereof that is capable or recognising the target site.

Abstract: The present invention describes methods of identifying altered recombinases and compositions thereof, wherein at least one amino acid is different from a parent, wild-type recombinase and the altered recombinase has improved recombination efficiency towards wild-type and/or pseudo att site sequences relative to the parent, wild-type recombinase. The present invention also includes methods of modifying the genomes of cells using the altered recombinases, including methods of site-specifically integrating a polynucleotide sequence of interest in a genome of a eucaryotic cell.

Abstract: The invention relates to the use of prokaryotic beta recombinase in eukaryotic cells, especially for transgenic work in eukaryotic cells. It also relates to the use of prokaryotic beta recombinase for site-specific intramolecular recombination between two six sites in eukaryotic cells. The use of the gene coding for beta recombinase for promoting the deletion of DNA sequences located between directly oriented six sites in mammalian cells and for catalysing site-specific resolution of DNA sequences in an extrachromosomal target introduced into an eukaryotic cell are also disclosed.

Abstract: Methods and DNA constructs are provided for detection and manipulation of a target eukaryotic gene whose expression is restricted to certain tissues or specialized cell types. The methods include transforming a cell with a first indicator component under the control of a promoter selected for its restricted expression in a particular cell or tissue. The cell is also transformed with a gene trap vector encoding a second indicator component. The cell is allowed to differentiate to produce specialized cell or tissue which is monitored for expression of both the first and second indicator components, thereby detecting a gene into which the trap vector has integrated which is expressed in the same cell or tissue type as the selected promoter.

Abstract: The invention provides methods for modulating a cellular process by contacting a cell in culture with a cell process-modifying molecule attached to a translocating polypeptide. For example, in one embodiment, a cell in culture is transfected with a target gene by contacting the cell in culture with a polynucleotide (that contains the target gene) attached to a translocating polypeptide. In another embodiment, expression of a target gene product in a cell in culture that contains a target gene under control of one or more regulatory elements is modulated by contacting the cell in culture with one or more regulatory agents attached to a translocating polypeptide. The one or more regulatory agents are translocated into the cell in culture and interact therein with the one or more regulatory elements to modulate expression of the target gene product by the cell.

Abstract: The present invention provides a process for preparing a retrovirus to be expressed at a high titer by specifically transferring a desired foreign gene into target cells. A pseudotyped retrovirus vector having a high titer can be prepared by transferring a DNA construction wherein a promoter, an loxP sequence, a VSV-G gene and a polyA addition signal are arranged in this order is transferred into cells carrying the retrovirus gag and pol gene expression systems, and then transferring a retrovirus vector containing the desired foreign gene thereinto, followed by treatment with a recombinase.

Abstract: Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).

Abstract: The present invention provides a strategy that allows for selective switching off of both transgene and viral gene expression in tissues where such expression is undesirable. The present invention employs a vector containing a tissue specific promoter that drives expression of Cre recombinase gene in tissue where transgene expression is undesirable. As a result of Cre recombinase expression, the same or another vector that expresses the transgene in that tissue will be cut by the action of the Cre recombinase into several pieces due to LoxP sites that are strategically placed within the vector backbone. Consequently, unwanted transgene as well as viral gene expression are prevented.

Abstract: According to the invention, there is provided a vector for the expression of immunoglobulin-cytokine fusion proteins in malignant B cells at least containing operably linked to each other (a) a region of at least 1.5 kb which is homologous to a region of the &mgr; intron or the &kgr; intron and which lacks a functional C&mgr; or C&kgr; enhancer or contains a non-functional C82 or C&kgr; enhancer; (b) at least one DNA sequence encoding a domain of an immunoglobulin or a part thereof; (c) a DNA sequence encoding a cytokine; and (d) a marker gene selectable in eukaryotic B cells and lacking a functional enhancer region wherein the expression of said marker following integration is controlled by the cellular C&mgr; or C&kgr; enhancer.

Abstract: A functionally active hyaluronan synthase having at least one modified amino acid residue therein as compared to a corresponding functionally active native hyaluronan synthase such that the functionally active hyaluronan synthase has an altered enzymatic activity as compared to the corresponding functionally active native hyaluronan synthase is disclosed. Methods of producing hyaluronic acid utilizing a recombinant host cell having an expression construct encoding the functionally active hyaluronan synthase with altered enzymatic activity are also disclosed.

Abstract: The present invention provides methods of site-specifically integrating a polynucleotide sequence of interest in a genome of a eucaryotic cell, as well as, enzymes, polypeptides, and a variety of vector constructs useful therefore. In the method, a targeting construct comprises, for example, (i) a first recombination site and a polynucleotide sequence of interest, and (ii) a site-specific recombinase, which are introduced into the cell. The genome of the cell comprises a second recombination site. Recombination between the first and second recombination sites is facilitated by the site-specific recombinase. The invention describes compositions, vectors, and methods of use thereof, for the generation of transgenic cells, tissues, plants, and animals. The compositions, vectors, and methods of the present invention are also useful in gene therapy techniques.

Abstract: Methods and compositions for the targeted integration of nucleotide sequences into a plant are provided. Particularly, the present invention is drawn to compositions comprising polynucleotide sequences having the following operably linked components: an intron, a nucleotide sequence of interest, and a terminator region, wherein the polynucleotide sequence comprises one or more recombination sites. The recombination sites are non-identical to one another and one of the recombination sites is contained within the intron.

Abstract: A method for generating and amplifying closed circular DNA having a specific sequence in vitro in a cell-free system is disclosed. Prior to the invention of this method, closed circular DNA could only be amplified in vivo in appropriate host cells. The essence of the method is the inclusion of a thermostable DNA ligase in a PCR reaction. This procedure is referred to as ligation-during-amplification (LDA), in which the fully extended DNA strands are ligated by the DNA ligase and used as templates for subsequent amplification. Closed circular DNA having a specific sequence can be selectively amplified exponentially by the use of two sequence-specific primers in the LDA reaction. In addition, one or more site-specific mutations can be introduced into a closed circular DNA by the use of one or more mutagenic primers in the LDA reaction. Various thermostable DNA polymerases and thermostable ligases can be used for LDA amplification.

Abstract: A method for engineering and utilizing large DNA vectors to target, via homologous recombination, and modify, in any desirable fashion, endogenous genes and chromosomal loci in eukaryotic cells. These large DNA targeting vectors for eukaryotic cells, termed LTVECs, are derived from fragments of cloned genomic DNA larger than those typically used by other approaches intended to perform homologous targeting in eukaryotic cells. Also provided is a rapid and convenient method of detecting eukaryotic cells in which the LTVEC has correctly targeted and modified the desired endogenous gene(s) or chromosomal locus (loci) as well as the use of these cells to generate organisms bearing the genetic modification.

Abstract: A novel rapid mutational analysis method for mapping protein epitopes is disclosed. This method has been used to identify the binding sites for 16 anti-CD2 and anti-CD4 monoclonal antibodies. The powerful, rapid, and simple method of the present invention allows isolation of a very large number of mutants, and is applicable to any intracellular or surface protein for which a cDNA and monoclonal antibodies are available. The present method is especially useful in ligand binding site studies for the design of new ligands and drugs.

Abstract: Methods for reducing the complexity of 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: The use of a circular vector DNA to produce a pharmaceutical agent for the treatment of mammals or humans by gene therapy wherein the vector DNA contains a selection marker gene and a DNA sequence that is heterologous for the vector which causes a modulation, correction or activation of the expression of an endogenous gene or the expression of a gene introduced into the cells of the mammal or the human by the vector DNA which is characterized in that the vector nucleic acid a) is amplified under selection pressure and cleaved in such a way that the said selection marker gene and the said heterologous DNA are present on separate DNA fragments, b) the DNA fragment which contains the heterologous DNA or both fragments are recircularized to form vectors, c) the DNA fragments are separated before or after the recircularization d) the recircularized DNA fragment which contains the heterologous DNA is isolated and e) the recircularized DNA fragment obtained in this manner is used to produce the pharmaceutical a

Abstract: Transgenic mice that produce high levels of humanized antibodies are described. Targeted gene replacement exchanges constant regions of the mouse immunoglobulin heavy and light chain genes with human genes, either through conventional gene targeting, or by use of the bacteriophage-derived Cre-loxP recombination system. The transgenic animals undergo antibody affinity maturation, and a class switch from the native immunoglobulin to the humanized form.