Abstract: The problems to be solved by the present invention are to provide: a recombinant varicella-zoster virus; a process for producing the same; a pharmacological composition containing a recombinant varicella-zoster virus; a vector containing a BAC vector sequence in the specific gene of a genomic gene of varicella-zoster virus; cells containing such a vector; a fragment capable of homologous recombination with a genome of varicella-zoster virus; a nucleic acid cassette containing the BAC vector sequence; and a multivalent vaccine. The above problems were solved by developing a process for producing a recombinant varicella-zoster virus, wherein the BAC vector sequence is inserted into a specific virus gene.

Abstract: Single AAV vector constructs for expression of an immunoglobulin molecule or fragment thereof and methods of making and using the same are described. The AAV vectors comprise a self-processing cleavage sequence between a first and second immunoglobulin coding sequence allowing for expression of a functional antibody molecule using a single promoter. The vector constructs may further include an additional proteolytic cleavage sequence which provides a means to remove the self processing peptide sequence from an expressed immunoglobulin molecule or fragment thereof. The vector constructs find utility in enhanced production of biologically active immunoglobulins or fragments thereof in vitro and in vivo.

Abstract: The invention provides a nucleic acid molecule comprising: (a) a nucleotide sequence as shown in SEQ ID No. 35; or (b) a nucleotide sequence which is the complement of SEQ ID No. 35; or (c) a nucleotide sequence which is degenerate with SEQ ID No. 35; or (d) a nucleotide sequence hybridising under conditions of high stringency to SEQ ID No. 35, to the complement of SEQ ID No. 35, or to a hybridisation probe derived from SEQ ID No. 35 or the complement thereof; or (e) a nucleotide sequence having at least 80% sequence identity with SEQ ID No. 35; or (f) a nucleotide sequence having at least 65% sequence identity with SEQ ID No. 35 wherein said sequence preferably encodes or is complementary to a sequence encoding a nystatin PKS enzyme or a part thereof.

Abstract: The CAMP factor gene of Streptococcus uberis (S. uberis) is described, as well as the recombinant production of CAMP factor therefrom. Also disclosed are chimeric CAMP factor constructs, including CAMP factor epitopes from more than one bacterial species. The CAMP factors and chimeras including the same can be used in immunogenic compositions for the prevention and treatment of bacterial infections.

Abstract: The present invention relates to a method for the cultivation of primary cells. The primary cells are cultivated in a serum free medium comprising a factor selected from the group consisting of growth factors and attachment factors. The method for the cultivation of primary cells may be one step in a method for the amplification of viruses, such as poxviruses. According to this latter method the primary cells are cultivated in a serum free medium comprising a factor selected from the group consisting of growth factors and attachment factors. The cells are then infected with the virus and the infected cells are cultivated in serum free medium until progeny virus is produced.

Abstract: The present invention provides minicircle nucleic acid vector formulations for use in administering to a subject, wherein the minicircle nucleic acid vectors include a polynucleotide of interest, a product hybrid sequence of a unidirectional site-specific recombinase, and are devoid of plasmid backbone bacterial DNA sequences. Also provided are methods of producing the subject formulations as well as methods for administering the minicircle nucleic acid vector formulations to a subject. The subject methods and compositions find use in a variety of different applications, including both research and therapeutic applications.

Abstract: The invention relates to newly identified polynucleotide sequences comprising a gene that encodes a novel oxidoreductase isolated from Aspergillus niger. The invention features the full length nucleotide sequence of the novel gene, the cDNA sequence comprising the full length coding sequence of the novel oxidoreductase as well as the amino acid sequence of the full-length functional protein and functional equivalents thereof. The invention also relates to methods of using these enzymes in baking and in dairy applications. Also included in the invention are cells transformed with a polynucleotide according to the invention and cells wherein a oxidoreductase according to the invention is genetically modified to enhance or reduce its activity and/or level of expression.

Abstract: Methods and compositions for cloning a donor DNA molecule into an acceptor vector at a predetermined location are described. The methods are based on homologous recombination mediated by in vitro treatment of the donor DNA and the acceptor vector with an enzyme cocktail containing an exonuclease and a single-stranded DNA binding protein.

Abstract: The present invention relates to novel polypeptides according to caroase 01-05 or any functional equivalents of any of them, suitable for use in a method for preparing a food products having increased whiteness, the use of the enzyme to increase whiteness of at least part of a food product, a process for preparing a food product wherein the enzyme is used and the food product obtained.

Abstract: Methods and kits for generating circular nucleic acids in a cell-free system, and uses for the generated circular nucleic acids are provided. The methods comprise in vitro amplification of a nucleic acid template comprising a recombination site to produce tandem repeat nucleic acid sequence, and employ a recombination protein to generate the circular nucleic acids from the tandem repeat nucleic acid sequence.

Abstract: The present invention provides methods of producing an isoprenoid or an isoprenoid precursor in a genetically modified host cell. The methods generally involve modulating the level of hydroxymethylglutaryl-CoA (HMG-CoA) in the cell, such that the level of HMG-CoA is not toxic to the cell and/or does not substantially inhibit cell growth, but is maintained at a level that provides for high-level production of mevalonate, IPP, and other downstream products of an isoprenoid or isoprenoid pathway, e.g., polyprenyl diphosphates and isoprenoid compounds. The present invention further provides genetically modified host cells that are suitable for use in a subject method. The present invention further provides recombinant nucleic acid constructs for use in generating a subject genetically modified host cell, including recombinant nucleic acid constructs comprising nucleotide sequences encoding one or more mevalonate pathway enzymes, and recombinant vectors (e.g., recombinant expression vectors) comprising same.

Abstract: The invention relates to a newly identified polynucleotide sequence comprising a gene that encodes a novel phospholipase isolated from Aspergillus niger. The invention features the full length nucleotide sequence of the novel gene, the cDNA sequence comprising the full length coding sequence of the novel phospholipase as well as the amino acid sequence of the full-length functional protein and functional equivalents thereof. The invention also relates to methods of using these enzymes in industrial processes and methods of diagnosing fungal infections. Also included in the invention are cells transformed with a polynucleotide according to the invention and cells wherein a phospholipase according to the invention is genetically modified to enhance or reduce its activity and/or level of expression.

Abstract: The invention relates to a method for obtaining a chromosomal locus for transgenesis of a multicellular eukaryotic organism, “MEO”, to a vector for transgenesis by homologous recombination of a MEO and to the use of such an vector for trans-genesis by homologous recombination of a MEO and to the transgenic MEO thus obtainable.

Abstract: Nucleic acid molecules which encode a branching enzyme from a bacterium of the genus Neisseria, vectors, host cell, plant cells and plants containing said nucleic acid molecules as well as starch obtainable from the plants described are described. Furthermore, an in-vitro method for producing ?-1,6-branched ?-1,4-glucans on the basis of sucrose and a combination of enzymes of an amylosucrase and a branching enzyme as well as the ?-1,6-branched ?-1,4-glucans obtainable by said method are described.

Abstract: The present invention relates to compositions and methods for enhancing the oncolytic activity of replication-competent, target cell-specific adenovirus vectors by modification of the E1A gene product. The target cell-specific replication-competent adenovirus vectors comprise a chimera of an adenovirus gene essential for replication, preferably an early gene, and the Androgen receptor (or a portion thereof) under the transcriptional control of a cell type-specific transcriptional regulatory element (TRE). By providing for cell type-specific transcription through the use of one or more cell type-specific TREs, the adenovirus vectors effect prostate-specific cytotoxicity due to selective replication.

Abstract: The present invention provides methods for a robust production of isoprenoids via one or more biosynthetic pathways. The invention also provides nucleic acids, enzymes, expression vectors, and genetically modified host cells for carrying out the subject methods. The invention also provides fermentation methods for high productivity of isoprenoids from genetically modified host cells.

Abstract: An attenuated feline recombinant herpesvirus 1 (FHV-1), which is prepared by identifying gene regions in the genome wherein inserted foreign genes can be expressed without affecting the replication of FHV-1 and has least two types of foreign nucleic acid sequences inserted thereinto, usable as a vector virus or a vaccine. In this attenuated recombinant FHV-1, at least two types of foreign genes are inserted in such a manner as allowing the expression into two different gene regions exerting no lethal effect on the proliferation of the virus in the feline herpesvirus 1 genome.

Abstract: Method of preparing 1-CreI meganuclease variants having a modified cleavage specificity, variants obtainable by said method and their applications either for cleaving new DNA target or for genetic engineering and genome engineering for non-therapeutic purposes. Nucleic acids encoding said variants, expression cassettes comprising said nucleic acids, vectors comprising said expression cassettes, cells or organisms, plants or animals except humans, transformed by said vectors.

Abstract: Provided is a method for constructing microRNA adenovirus expression plasmids comprising severe hepatitis related genes of hfgl2 (Human Fibrinogen-like protein 2) prothrombinase, Fas and TNFR1 (tumor necrosis factor receptor 1), and the specificity and effectivity of microRNA interference plasmids are detected at cellular level so that a combined use of three microRNA adenovirus expression plasmids for the treatment of severe hepatitis can be achieved. According to the present invention, pAd/CMV/V5-DEST vectors and pcDNA expression plasmid of hfgl2, hFas and hTNFR1 are used to construct pAd-hfgl2, pAd-hFas and pAd-hTNFR1 by means of Gateway technology.

Abstract: This invention relates to a human artificial chromosome (HAC) vector carrying a human chromosome-derived centromere, a subtelomere sequence, and a telomere sequence, to a human cell medicine or human cells comprising the HAC vector, to methods for preparing the HAC vector and human cells, and to methods for producing a therapeutic protein using the HAC vector.

Abstract: The present invention provides compositions, methods, and kits for covalently linking nucleic acid molecules. The methods include a strand invasion step, and the compositions and kits are useful for performing such methods. For example, a method of covalently linking double stranded (ds) nucleic acid molecules can include contacting a first ds nucleic acid molecule, which has a topoisomerase linked to a 3? terminus of one end and has a single stranded 5? overhang at the same end, with a second ds nucleic acid molecule having a blunt end, such that the 5? overhang can hybridize to a complementary sequence of the blunt end of the second nucleic acid molecule, and the topoisomerase can covalently link the ds nucleic acid molecules. The methods are simpler and more efficient than previous methods for covalently linking nucleic acid sequences, and the compositions and kits facilitate practicing the methods, including methods of directionally linking two or more ds nucleic acid molecules.

Abstract: The present invention concerns the technical field of nucleic acids and expression-optimized nucleic acids. The present invention concerns especially nucleic acids comprising a mutated foamy viral envelope gene encoding a foamy viral envelope polypeptide, which comprises a leader peptide (LP), a surface unit (SU) and a transmembrane domain (TM). The present invention also relates to modified polypeptides encoded by these nucleic acids. Furthermore, the present invention regards a method for preparing pseudotyped vector particles as well as a method for treating a genetic disorder comprising administering a nucleic acid or a polypeptide encoded by that nucleic acid.

Abstract: The present invention provides an improved system for linking nucleic acids to one another. In particular, the present invention provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules. The product molecules need not be cleaved with restriction enzymes in order to undergo such ligation. In preferred embodiments of the invention, the DNA product molecules are produced through iterative DNA synthesis reactions, so that the product molecules are amplified products. The invention further provides methods for directed ligation of product molecules (i.e., for selective ligation of certain molecules within a collection of molecules), and also for methods of exon shuffling, in which multiple different product molecules are produced in a single ligation reaction. Preferred embodiments of the invention involve ligation of product molecules encoding functional protein domains, particularly domains naturally found in conserved gene families.

Abstract: It is an object of the present invention to provide a simple and efficient means capable of evaluating the VH/VL interaction without expressing/purifying VH and VL. The present invention provides a recombinant vector comprising: (i) a nucleotide sequence which can express a hetero-assembly composed of two types of fusion proteins wherein heavy chain variable region (VH) and light chain variable region (VL) of antibody are respectively fused with mutually associable first polypeptide and second polypeptide, by means of secretion, or in a form of a fusion protein tethered to a phage coat protein; and (ii) a restriction enzyme recognition sequence at two sites within, or in a vicinity of, a nucleotide sequence encoding said first polypeptide or second polypeptide.

Abstract: The invention discloses the production of double stranded DNA (dsDNA) vectors capable of delivering nucleic acids, including cDNA, antisense, ribozyme, and small interference RNA into cells. The invention also describes nucleic acid constructs as well as methods for the production of the dsDNA vectors.

Abstract: The present invention provides HIV-derived lentivectors which are multiply modified to create highly safe, efficient, and potent vectors for expressing transgenes for gene therapy. The lentiviral vectors comprise various combinations of an inactive central polypurine tract, a stuffer sequence, which may encode drug susceptibility genes, and a mutated hairpin in the 5? leader sequence that substantially abolishes replication. These elements are provided in conjunction with other features of lentiviral vectors, such as a self-inactivating configuration for biosafety and promoters such as the EF1? promoter as one example. Additional promoters are also described. The vectors can also comprise additional transcription enhancing elements such as the wood chuck hepatitis virus post-transcriptional regulatory element.

Abstract: This invention provides expression vectors for a ribonucleic acid (RNA) molecule comprising a double-stranded region of random sequence, sets and libraries of same, methods of generating same, and methods for identifying an RNA therapeutic or RNA molecule that has an ability to affect a biological parameter, for identifying a drug target for a disease or disorder of interest, and for identifying a variant of an RNA molecule that has an altered ability to affect a biological parameter of interest.

Abstract: The present disclosure generally relates to processes for preparing highly pure plasmid compositions. The processes generally involve treating a composition comprising plasmid DNA with a polypeptide to digest colanic acid. The treated plasmid DNA is then separated from the treated composition.

Abstract: The invention relates to a system for stable maintenance of a plasmid, to host cells for use in this system and to methods of using the system to obtain a plasmid useful in medical applications. In particular, the invention provides transformed host cell containing: i) a chromosomal gene which inhibits cell growth; and ii) a plasmid encoding an antisense sequence, wherein the antisense sequence encoded by the plasmid inhibits the action of the chromosomal gene, thereby permitting cell growth and a method for stable maintenance of a plasmid in a host cell in vivo.

Abstract: The functional analysis of genes frequently requires the manipulation of large genomic regions. A yeast-bacteria shuttle vector is described that can be used to clone large regions of DNA by homologous recombination. Also described is a method for isolating entire genomes, including chloroplast genomes, or large portions thereof, and manipulating the same. Also described are methods for determining minimal genomes, minimal pathway requirements, and minimal organelle genomes.

Abstract: Herein is described a modified viral vector comprising: a coat protein modified, for example by the addition of a cysteine residue, such that the modified viral vector yields less soluble virus relative to that from an unmodified viral vector upon extraction of plant material infected with the modified viral vector, thereby facilitating purification of a recombinant protein expressed from the modified viral vector. Also described is a method of reducing viral coat protein impurities during purification of a recombinant protein, a method of biocontainment for a recombinant viral vector, and a method of generating virus inoculum for the modified viral vector.

Abstract: A protein expression vector that expresses large quantities of recombinant proteins under anoxic or microaerobic conditions by inducing expression with nitrate. The vector backbone is pUC19 and protein expression is driven by the E. coli flavohemoglobin promoter, which is inducible by nitrate, nitrite, or nitric oxide under conditions of low oxygen. The Nde1 site of pUC19 has been destroyed by filling in with Klenow fragment and religating the vector. An Nde1 site in the promoter provides an in-frame start methionine and a standard polylinker is available for ease of subcloning. The vector is named pANX for Plasmid ANaerobic eXpression.

Abstract: The present invention is directed to compositions and methods for nucleic acid identification and detection. Compositions and methods of the present invention include template nucleic acids with stabilizing sequences. The present invention also includes concatemers formed from template nucleic acids that have stabilizing sequences, arrays of such concatemers, as well as methods for identifying and detecting sequences of such concatemers.

Abstract: A method of stimulating homologous recombination by creating at least one nick in a targeted polynucleotide sequence. Wherein nonhomologous recombination is suppressed resulting in increasing the ratio of targeted to nontargeted events. A method of increasing double strand break-initiated gene targeting by inducing a nick in a targeted polynucleotide sequence, wherein overall recombination levels are increased. A method of increasing homologous recombination employing a recombinase that releases the ends in living cells by stimulating homolgous recombination to higher levels than those attainable with standard nucleases. A composition for stimulating homologous recombination including a nicking mechanism for creating nicks in a polynucleotide, wherein the nicking mechanism stimulates homologous recombination. A composition for stimulating homologous recombination including a nicking endonucleases. Various kits for stimulating homologous recombination.

Abstract: Embodiments of the invention include E1 expressing cell lines that can be used in a variety of methods for production of an E1 defective adenovirus. In certain aspects a cell of the invention can be adapted to various culture conditions, e.g., suspension culture in serum free medium. In a further aspect, the cell lines allow isolation and subculture of E1-deleted recombinant adenoviruses in an environment free of replication competent adenovirus (RCA).

Abstract: The present invention provides a method for constructing recombinant translationally coupled operons, a method for producing useful metabolites using the bacterium containing the coupled operons, and a method for monitoring gene expression.

Abstract: This invention relates to the use of a polypeptide in the production of an immunostimulatory agent, said polypeptide comprising a sequence corresponding to the EDA domain of fibronectin, a fragment of the EDA domain which can bind to TLR4 or a variant of said EDA domain or a fragment which can bind to TLR4 and which has a homology of more than 70% with any form or natural fragment of the EDA domain. The invention also relates to the production methods and applications of said agent.

Abstract: The invention provides a novel method of vaccination of an animal of the felidae family against feline leukemia. The FeLV recombinant vaccine based on viral vector with the aid of a liquid jet needle-free injector can result in distribution of the vaccine essentially in the dermis and the hypodermis of the animal.

Abstract: Aspects described and claimed herein provide methods to insert multiple DNA adaptors into a population of circular target DNAs at defined positions and orientations with respect to one another. The resulting multi-adaptor constructs are then used in massively-parallel nucleic acid sequencing techniques.

Abstract: The invention provides an adenovirus that expresses a constitutively activated form of human NF-?B activating kinase (IKK). Co-administration of the adenovirus expressing the constitutively activated form of IKK (Ad-IKK?) and an antigen confers an enhanced immune response against the antigen relative to co-administration of the antigen and adenovirus that do not express IKK. Optionally, the adenovirus comprises one or more heterologous nucleic acid sequences that may encode, for example, a biologically active product such as the antigen of an infectious agent (e.g., bacterial, fungal, etc.) or a tumor antigen. Accordingly, the adenovirus is useful for inducing desired immune responses such as those against infectious agents and cancer cells.

Abstract: A method of producing a packaged parvovirus vector, the method comprising: (a) providing an insect cell; (b) introducing into the insect cell one or more vectors comprising nucleotide sequences encoding: (i) a transgene flanked by TRs; and (ii) baculovirus packaging functions comprising Rep components and Cap components sufficient to result in packaging of infective parvovirus particles, wherein VP1 is supplemented relative to VP2 and VP3 sufficient to increase the production of infectious viral particles; and (c) introducing into the cell a nucleic acid encoding baculovirus helper functions for expression in the insect cell; (d) culturing the cell under conditions sufficient to produce the infectious packaged parvovirus vector.

Abstract: Compositions and methods are provided for decreasing blood glucose levels in an animal or for preventing or delaying the onset of a rise in blood glucose levels in an animal, comprising administering to the animal an antisense inhibitor of PTP1B expression in combination with at least one glucose-lowering drug. The present invention is also directed to compositions and methods for improving insulin sensitivity in an animal or for preventing or delaying the onset of insulin resistance in an animal. Also provided are compositions and methods for treating or preventing a metabolic condition in an animal. The metabolic condition may be, e.g., diabetes or obesity.

Abstract: The present invention describes a method for producing synthetic nucleotide sequences which provides the assembly of DNA sequences, thus providing the obtention of genes, chromosomes and even whole qenomes. The method of the present invention makes use of the technique known as Polymerase Chain Reaction (PCR) but wherein no preexisting nucleic acid template is needed, being therefore an approach with minimum limitations and broad use. This method provides means for obtaining products with high industrial value, for the design and development of immunotherapeutic agents, recombinant enzymes, drugs, including the development of vaccines, gene therapy, and in applications in agriculture and environment.

Abstract: The present invention is a method for the purification of plasmid DNA comprising to provide a composition comprising a first polymer having inverse solubility characteristics and a second polymer immiscible in the first polymer; contacting said solution with an aqueous solution comprising plasmid DNA; providing phase separation and isolating the aqueous phase; and increasing the temperature of the isolated phase to a temperature above the cloud point of the first polymer and below the temperature where plasmid DNA is degraded and subsequently isolating the aqueous phase so formed. The invention also encompasses a kit for purification of plasmid DNA as described above.

Abstract: The present invention is directed to methods and compositions for acquiring nucleotide sequence information of target sequences using adaptors interspersed in target polynucleotides. The sequence information can be new, e.g. sequencing unknown nucleic acids, re-sequencing, or genotyping. The invention preferably includes methods for inserting a plurality of adaptors at spaced locations within a target polynucleotide or a fragment of a polynucleotide. Such adaptors may serve as platforms for interrogating adjacent sequences using various sequencing chemistries, such as those that identify nucleotides by primer extension, probe ligation, and the like. Encompassed in the invention are methods and compositions for the insertion of known adaptor sequences into target sequences, such that there is an interruption of contiguous target sequence with the adaptors. By sequencing both “upstream” and “downstream” of the adaptors, identification of entire target sequences may be accomplished.

Abstract: The present invention discloses a method of producing polypeptides, including insulinotropic GLP-1 (7-36) polypeptide and/or GLP-1 analogs, by ligating genes in a tandem way. Also disclosed are the recombinant polypeptides produced by such a method. Using the method of this invention, 1 to 32 copies of GLP-1 (7-36) and/or GLP-1 analog genes may be expressed in tandem and the desired polypeptide can be obtained after cleavage of a fusion protein and further processes of separation and purification thus making possible the production of recombinant polypeptides, including recombinant GLP-1 (7-36) and/or GLP-1 analogs on a large scale, at a significantly reduced production cost.

Abstract: The functional analysis of genes frequently requires the manipulation of large genomic regions. A yeast-bacteria shuttle vector is described that can be used to clone large regions of DNA by homologous recombination. Also described is a method for isolating entire genomes, including chloroplast genomes, or large portions thereof, and manipulating the same. Also described are methods for determining minimal genomes, minimal pathway requirements, and minimal organelle genomes.

Abstract: A method for the production of fungus resistant transgenic plants, plant cells or plant tissue comprising the introduction of an Ab, rAb, rAb fragment or fusion or vector of the invention or the vectors of the composition of the invention into the genome of a plant, plant cell or plant cell tissue and a transgenic plant cell comprising stably integrated into the genome a polynucleotide or vector of the invention or the vectors of the composition of the invention.

Abstract: The present invention is directed to a method to diversify the chemical composition of proteins produced in vivo comprising the step of disabling, particularly by mutagenesis, the editing function of one of its aminoacyl tRNA synthetases. The present invention is also directed to nucleic acid sequences encoding such mutated aminoacyl tRNA synthetases having their editing site mutated and capable of mischarging its cognate tRNA with a noncanonical amino acid. Also described herein is an improved method for obtaining transformed cells capable of synthetizing in vivo proteins comprising at least a noncanonical amino acid and their use for the production of such proteins.

Abstract: Functional shRNA is produced from an expression vector prepared by selecting a two primer design in which the primers are less than about 50 nucleotides in length, annealing and extending the primers using primer extension, digesting the primer extension product and inserting the digestion product into a suitable vector. When the shRNA vectors are inserted into a cell, shRNA transcribed from the vectors modulates gene activity within the cell.