Abstract: The present invention relates to oncolytic adenoviruses having therapeutic applications. Recombinant chimeric adenoviruses, and methods to produce them are provided. The chimeric adenoviruses of the invention comprise nucleic acid sequences derived from adenoviral serotypes classified within the subgroups B through F and demonstrate an enhanced therapeutic index.

Abstract: In one embodiment of the invention, a method is provided for preparing plasmid from host cells which contain the plasmid, comprising: (a) providing a plasmid solution comprised of unligatable open circular plasmid; (b) reacting the unligatable open circular plasmid with one or more enzymes and appropriate nucleotide cofactor(s), such that unligatable open circular plasmid is converted to 3?-hydroxyl, 5?-phosphate nicked plasmid; (c) reacting the 3?-hydroxyl, 5?-phosphate nicked plasmid with a DNA ligase and DNA ligase nucleotide cofactor, such that 3?-hydroxyl, 5?-phosphate nicked plasmid is converted to relaxed covalently closed circular plasmid; and (d) reacting the relaxed covalently closed circular plasmid with a DNA gyrase and DNA gyrase nucleotide cofactor, such that relaxed covalently closed circular plasmid is converted to negatively supercoiled plasmid. In other embodiments, DNA gyrase is replaced with reverse DNA gyrase or reaction (d) is not performed.

Abstract: The invention relates to a method for engineering a nucleic acid for expression in the presence of Kid/PemK endoribonuclease comprising (i) screening the nucleotide sequence of the nucleic acid for the sequence UUACU or TTACT (ii) mutating said sequence such that there are no longer any occurrences of UUACU or TTACT. The invention also relates to a method of making a ribonucleic acid resistant to Kid/PemK endoribonuclease, said method comprising (a) providing a nucleic acid; (b) screening the nucleic acid for the nucleotide sequence UUACU or TTACT; (c) mutating said sequence such that there are no longer any occurrences of UUACU or TTACT; wherein when the nucleic acid of (a) is a deoxyribonucleic acid, said method further comprises (d) transcribing said deoxyribonucleic acid to produce ribonucleic acid. The invention also relates to vectors and uses of purified or recombinant Kid/PemK endoribonucleases.

Abstract: Provided herein are methods, compositions, and kits useful for molecular cloning of, for example, blunt-ended DNA molecules using DNA topoisomerase. In certain embodiments, the methods comprise combining into a mixture a first polynucleotide, wherein said first polynucleotide comprises an origin of replication, a selectable marker, two topoisomerase recognition sequences, and two nicking agent recognition sequences, wherein each of said two topoisomerase recognition sequences is within 0-50 nucleotides of at least one of the nicking agent recognition sequences and wherein each of said two nicking agent recognition sequences is nicked, with a sequence-specific topoisomerase and a second polynucleotide, wherein said second polynucleotide comprises a 5? hydroxyl on each end of the polynucleotide; and transforming the mixture into a host organism, thereby cloning the second polynucleotide.

Abstract: An isolated nucleic acid molecule encoding a human DNA repair enzyme, MED1, is disclosed. Like other mismatch repair genes which are mutated in certain cancers, MED1, encoding nucleic acids, proteins and antibodies thereto may be used to advantage in genetic or cancer screening assays. MED1, which recognizes and cleaves DNA, may also be used for the diagnostic detection of mutations and genetic variants.

Abstract: The present invention comprises a copper-inducible promoter system of yeast for the controlled expression of recombinant genes. The invention comprises a promoter element comprising at least two tandemly-repeated MRE sequences and a minimal TATA element from the 35S-promoter of the cauliflower mosaic virus.

Abstract: The present invention provides an expression vector comprising genes encoding OmpF of E. coli and a desired protein, E.coli transformed with the expression vector, and a method for extracellular production of desired proteins by employing the same. The recombinant expression vector of the invention comprises an ampicillin-resistance gene, the OmpF promoter and the OmpF gene. In accordance with the invention, a desired protein can be produced extracellularly by a simpler method than conventional methods such that: secretory production of OmpF fusion protein begins simultaneously with growth of the cells through constitutive expression employing an OmpF promoter, and as the concentration of cells increases, the amount of secretory production of the protein also increases continuously. Therefore, desired proteins can be produced in large quantities by a high concentration culture of cells.

Abstract: A vector for transformation into a host cell is described comprising a toxic gene encoding a product that is lethal to the host cell, wherein the toxic gene comprises: an essential sequence region whose integrity is necessary in order for the encoded toxic gene product to be lethal to the host cell; an inessential sequence region whose integrity is not essential in order for the encoded toxic gene product to be lethal to the host cell; a regulatory sequence inserted in-frame into the inessential sequence region; and a cloning site within the essential sequence region for insertion of a nucleic acid sequence, wherein the regulatory sequence and the cloning site are positioned so as to allow the regulatory sequence to be operably linked to a nucleic acid sequence when the nucleic acid sequence is inserted into the cloning site.

Abstract: The instant invention provides methods and compositions for generating recombinant adenoviral vectors. The invention also provides kits comprising for the generation of recombinant adenoviral vectors.

Abstract: The invention belongs to the field of animal health, in particular equine diseases caused by equine herpesvirus (EHV). The invention relates to artificial chromosomes comprising the genome of equine herpesviruses, methods of producing attenuated or virulent EHV with or without the insertion of foreign genes, EHV obtainable with said methods and pharmaceutical compositions comprising said viruses.

Abstract: The invention relates to a gene isolated from E. coli, dep, which confers resistance to the antibacterial activity of 4,5-dihydroxy-2-cyclopenten-1-one (DHCP). The invention further relates to the putative protein encoded by dep, which is a hydrophobic, transmembrane efflux protein specific for DHCP. The invention further relates to plasmids containing the dep gene, and to bacterial cells expressing dep. Furthermore, the invention provides applications for use in conferring resistance to antibacterial activity in organisms. The dep gene can be used to identify compounds which inhibit the efflux activity responsible for the resistance to DHCP or to compounds which are functionally equivalent to DHCP.

Abstract: The invention relates to virus like particles, their preparation and their use in pharmaceutical screening and functional genomics. The invention further provides a variety of assay formats to be used with said virus like particles.

Abstract: The present invention relates to recombinant raccoon poxvirus vectors that express the rabies virus glycoprotein gene at the hemagglutinin (ha) locus of the poxvirus genome or express the glycoprotein gene of the same or different rabies strains at the thymidine kinase (tk) and the hemagglutinin (ha) loci of the poxvirus genome, and their use as adjuvant-free vaccines. The raccoon poxvirus vector comprises the nucleic acid molecules encoding the glycoprotein of a Challenge Virus Standard rabies strain inserted and expressed at the tk locus of the poxvirus genome and of a Pasteur-Paris rabies strain inserted and expressed at the ha locus of the poxvirus genome. The vaccine may optionally contain a mixture of additional feline and canine antigens for immunization of animals. Also disclosed are methods for inducing an immune response to rabies in a mammal by administering to the mammal an effective immunizing amount of the vaccine of the invention.

Abstract: The invention relates to a method for the production of vectors which, following transfection thereof in eukaryotic cells, are suitable for targeted inhibition of the formation of defined proteins therein by RNA interference. The method for the production of such vectors does not include any PCR steps. It is a three-step procedure in a single reaction vessel and can be carried out within a few hours. Thus, a method is provided which allows very easy testing of a wide variety of siRNA sequences for their functionality within a very short time. Screening processes utilizing the rapid and uncomplicated production of vectors with the aid of said kit can be performed in a cost- and time-saving manner. Another advantage of vectors thus produced is their small size which, among other things, facilitates transfection.

Abstract: Materials and Methods are provided for replacing one or more amino acids in a polypeptide with an amino acid of choice to form mutant proteins. Both naturally and non-naturally occurring amino acids can be inserted. A population of mutant proteins can be created in which an amino acid residue has replaced an existing residue at random locations along the primary sequence of the protein. The provided techniques allow for the study of proteins and development of proteins with improved functionalities.

Abstract: The present invention is drawn to fusion proteins comprising a Receptor for Advanced Glycation Endproducts (RAGE) and an immunoglobulin element. The invention also encompasses methods of treating a condition characterized by activation of an inflammatory cytokine cascade comprising administering such fusion proteins. The invention is also drawn to nucleic acids encoding the fusion proteins, as well as vectors and cells comprising such nucleic acids.

Abstract: A continuous process is described for the production of microbial plasmid DNA for use in biopharmaceutical and biotechnological applications. The process consists of: first growing microbial cells containing a plasmid at a reduced temperature in a continuous stage; followed by a second plasmid induction continuous culture stage with an increased temperature, with a residence time that allows accumulation of the plasmid product. A hold step at a reduced temperature after fermentation further increases the yield of plasmid product. The method enables production of a large quantity of highly purified plasmid DNA from a small bioreactor over time.

Abstract: The present invention provides duplexed parvovirus vector genomes that are capable under appropriate conditions of forming a double-stranded molecule by intrastrand base-pairing. Also provided are duplexed parvovirus particles comprising the vector genome. Further disclosed are templates and methods for producing the duplexed vector genomes and duplexed parvovirus particles of the invention. Methods of administering these reagents to a cell or subject are also described. Preferably, the parvovirus capsid is an AAV capsid. It is further preferred that the vector genome comprises AAV terminal repeat sequences.

Abstract: Lentiviral vectors modified at the 5? LTR or both the 5? and 3? LTR's are useful in the production of recombinant lentivirus vectors. Such vectors can be produced in the absence of a functional tat gene. Multiple transformation of the host cell with the vector carrying the transgene enhances virus production.

Abstract: The invention relates to the nucleic acid and polypeptide sequences of three novel human Ron-related gene variants (Ron-V1, Ron-V2, and Ron-V3). The invention also provides a process for producing the polypeptides of the variants, as well as uses for the nucleic acid, polypeptide and antibodies to same in diagnosing human breast carcinoma, breast adenocarcinoma, cervix epidermoid carcinoma, cervix epitheloid carcinoma, colon adenocarcinoma, urinary bladder carcinoma, prostate carcinoma, esophagus epidermoid carcinoma and esophagus carcinoma.

Abstract: Improved vaccines and methods of using the same are disclosed Immunosuppressive compositions for treating individuals who have autoimmune diseases or transplants and methods of using the same are disclosed.

Abstract: The invention provides methods for evolving a polynucleotide toward acquisition of a desired property. Such methods entail incubating a population of parental polynucleotide variants under conditions to generate annealed polynucleotides comprising heteroduplexes. The heteroduplexes are then exposed to a cellular DNA repair system to convert the heteroduplexes to parental polynucleotide variants or recombined polynucleotide variants. The resulting polynucleotides are then screened or selected for the desired property.

Abstract: The present invention relates to retroviral vectors, particularly lentiviral vectors, pseudotyped with Sindbis envelope and targeted to specific cell types via a targeting moiety linked to the envelope.

Abstract: This invention relates to polynucleotides, (herein referred to as “BASB206 polynucleotide(s)”), polypeptides encoded by them (referred to herein as “BASB206” or “BASB206 polypeptide(s)”), recombinant materials and methods for their production. In another aspect, the invention relates to methods for using such polypeptides and polynucleotides, including vaccines against bacterial infections. In a further aspect, the invention relates to diagnostic assays for detecting infection of certain pathogens.

Abstract: The present application is directed to a method for performing a bisulfite reaction to determine methylation positions in a nucleic acid, i.e. methylated and non-methylated cytosines, whereby the nucleic acid is incubated in a solution comprising the nucleic acid for a time period of 1.5 to 3.5 hours at a temperature between 70 and 90° C., whereby the concentration of bisulfite in the solution is between 3 M and 6.25 M and whereby the pH value of the solution is between 5.0 and 6.0 whereby the nucleic acid, i.e. the cytosine bases in the nucleic acid, are deaminated. Then the solution comprising the deaminated nucleic acid is desulfonated and preferably desalted. The application is further related to a solution comprising bisulfite with a certain pH and uses thereof as well as a kit comprising the solution.

Abstract: The application relates for method for cloning the gene comprising the steps of: 1. Providing a replication-deficient baculovirus vector, 2. Providing a rescue vector comprising (a) nucleic acid sequence which is capable of restoring replication in the replication-deficient baculovirus vector and (b) at least one gene to be cloned; 3. Causing the replication-deficient baculovirus vector and rescue vector to recombine to produce a replication-enabled baculovirus vector comprising the at least one gene to be cloned; and 4. Growing the replication-enabled baculovirus vector within a suitable invertebrate cell, such as an insect cell. Preferably the baculovirus vector is based upon AcMNPV. Also disclosed are replication-deficient baculovirus vectors, rescue vectors, cells containing such vectors and kits comprising such vectors.

Abstract: HCV variants are described. The variants include polynucleotides comprising non-naturally occurring HCV sequences and HCV variants that have a transfection efficiency and ability to survive subpassage greater than HCV that have wild-type polyprotein coding regions. Expression vectors comprising the above polynucleotides and HCV variants are also described, as are the provision of cells and host cells comprising the expression vectors. Methods for identifying a cell line that is permissive for infection with HCV are also provided, as are methods for identifying HCV variant polynucleotides with increased transfection efficiencies. Additionally, methods for identifying one or more HCV sequence mutations that provide drug-resistant HCV variants are disclosed.

Abstract: The present invention relates to a method of mutagenesis for introducing mutations into a molecule and, in particular, to methods that may be applied to populations of molecules for the generation or screening of libraries involving the mutation of multiple molecules.

Abstract: Vector preparations and cloning constructs suitable for use in cloning are provided. Vector preparations are double-stranded DNA molecules having two 3? termini, each terminus having a single base pair overhang that is capable of hybridizing to a single base pair overhang on a double stranded polynucleotide sequence to be cloned. The overhang of the vector preparation is suitably a dCMP and the overhang of the polynucleotide sequence to be cloned is suitably a dGMP. In other embodiments, the overhang of the polynucleotide sequence to be cloned is any ddNTP and the corresponding overhang of the vector preparation is any base that pairs to the ddNTP. The latter embodiment is particularly suited to preparing recombinant molecules having only a single insert. Methods of cloning, methods of preparing libraries of recombinant molecules and kits for carrying out the methods are also provided.

Abstract: The present invention provides a simple method for producing a dumbbell-shaped DNA.

Abstract: Disclosed are recombinant human tyrosinase, which the transmembrane domain of human tyrosinase is removed and a method of producing the same. The method includes expressing human tyrosinase gene, which C-terminal transmembrane domain of human tyrosinase is removed, and separating and purifying the expressed recombinant human tyrosinase. The method according to the present invention have effects on mass-producing the recombinant human tyrosinase having a good enzyme activity without forming insoluble inclusion body, a simple purifying process and a improved purifying yield.

Abstract: The present invention relates to a method for producing a modified foreign chromosome(s) or a fragment(s) thereof, which comprises the steps of: (a) preparing a microcell comprising a foreign chromosome(s) or a fragment(s) thereof, and transferring said foreign chromosome(s) or a fragment(s) into a cell with high homologous recombination efficiency through its fusion with said microcell; (b) in said cell with high homologous recombination efficiency, inserting a targeting vector by homologous recombination into a desired site of said foreign chromosome(s) or a fragment(s) thereof, and/or a desired site of a chromosome(s) derived from said cell with high homologous recombination efficiency, thereby marking said desired site; and (c) in said cell with high homologous recombination efficiency, causing deletion and/or translocation to occur at the marked site of said foreign chromosome(s) or a fragment(s) thereof.

Abstract: The invention provides methods and compositions for in vivo incorporation of unnatural amino acids. Also provided are compositions including proteins with unnatural amino acids.

Abstract: The present invention relates to a novel system for expressing nitrile hydratase. In this system, the nitrile hydratases, which are composed of subunits, are formed such that the respective subunits are located on different plasmids and are expressed simultaneously in E. coli.

Abstract: This invention provides compositions and methods for generating components of protein biosynthetic machinery including orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases. Methods for identifying orthogonal pairs are also provided. These components can be used to incorporate unnatural amino acids into proteins in vivo.

Abstract: This invention provides a stress-inducible promoter that effectively functions in monocotyledonous plants such as rice, and environmental stress-tolerant plants using the promoter. Such promoter is derived from rice and consists of the following DNA (a) or (b): (a) DNA that consists of the nucleotide sequence as shown in SEQ ID NO: 1 or 10; or (b) DNA that hybridizes under stringent conditions with DNA consisting of a nucleotide sequence that is complementary to the DNA consisting of the nucleotide sequence as shown in SEQ ID NO: 1 or 10 and that expresses stress-inducible promoter activity. Such environmental stress-tolerant plant has had such promoter introduced therein.

Abstract: The present invention is directed to bacteriophage therapy, using methods which enable the bacteriophage to delay inactivation by any and all parts of the host defense system (HDS) against foreign objects. The HDS normally reduces the number of bacteriophage in an animal, which decreases the efficiency of the bacteriophage in killing the host bacteria present during an infection. Disclosed is a method of producing bacteriophage modified for anti-HDS purposes by physico-chemical alteration of the bacteriophage surface proteins, so that the altered bacteriophage remain active in the body for longer periods of time than the unmodified bacteriophage.

Abstract: Disclosed are the methods for producing recombinant viruses using site-specific recombination in vitro. In the present invention, circular viral genomic DNAs are digested with restriction enzymes to generate a linear form viral genomic DNAs flanked by site-specific recombination sites, and then are subjected to site-specific recombination with the desired genomic materials flanked by site-specific recombination sites in vitro. According to the present invention, since the site-specific recombination mixture can be applied to host cells without further procedures of selecting the desired recombinant viral genomic DNAs, it is possible to obtain numerous recombinant viruses rapidly at the same time. Thus, the present invention can be used as a high throughput system for generating and screening hundreds or thousands of recombinant viruses.

Abstract: The invention relates to a structural protein of adeno-associated virus (AAV) which comprises at least one mutation which brings about an increase in the infectivity of the virus.

Abstract: Disclosed herein are new defective Sindbis viral vectors made from wild type Ar-339 Sindbis virus, with differences in replicase and envelope proteins between JT vectors and consensus Sindbis virus sequences, and also between JT and Ar-339 vectors. Also disclosed are plasmids used for the production of the vectors, methods for producing the vectors, methods for treating mammals suffering from tumors and pharmaceutical formulations for use in the treatment methods.

Abstract: Large circular (LC)-sense molecules in an array is disclosed. The LC-sense molecules array is combined with cDNA hybridization to detect differences in expression profile between different cells. LC-sense molecules were purified from nonredundant clones with recombinant phagemid and arrayed onto silanized slide glasses. By hybridization of LC-sense array with Cy3 or Cy5-labelled cDNA preparations at 60° C., 29 up-regulated and 6 down-regulated genes in cancerous liver tissue were detected.

Abstract: The present invention provides means to modify the tropism of recombinant adenoviral vectors using genetic methods to alter the adenoviral fiber cell-binding protein. The present invention generates an adenovirus with modified fiber gene such that novel tropism is achieved. This recombinant adenovirus has a fiber gene modified in the HI loop domain.

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: A method for providing an adenovirus from a serotype which does not grow efficiently in a desired cell line with the ability to grow in that cell line is described. The method involves replacing the left and right termini of the adenovirus with the corresponding termini from an adenovirus which grow efficiently in the desired cell line. At a minimum, the left terminus spans the 5? inverted terminal repeat, the left terminus spans the E4 region and the 3? inverted terminal repeat. The resulting chimeric adenovirus contains the internal regions spanning the genes encoding the penton, hexon and fiber from the serotype which does not grow efficiently in the desired cell. Also provided are vectors constructed from novel simian adenovirus sequences and proteins, host cells containing same, and uses thereof.

Abstract: The present invention is related to a family of novel spatiotemporally active Rubisco promoters (SEQ ID NO: 1, 2, 3) obtainable from light grown Brassica seedlings. Furthermore the invention is related to transgene expression in specific plant organs or at specific stages of plant development. DNA constructs and expression cassettes comprising at least one of the promoter sequences functionally fused in frame with genes encoding desired gene products are disclosed. Seeds from transformed homologous and heterologous plants and from subsequent generation of the transformed plants are collected and used for efficient production of desired gene products, especially in contained conditions.

Abstract: The invention provides novel yeast promoters useful for controlling the expression of homologous and heterologous nucleic acid molecules in yeast cells. The yeast promoters are induced by a fermentable carbon source, such as glucose, or a non-fermentable carbon source, such as ethanol, or both. Therefore, expression of nucleic acid molecules encoding a polypeptide under the control of the novel yeast promoters may be regulated by varying the level of a fermentable carbon source, or a non-fermentable carbon source, or both.

Abstract: The present invention relates to compositions comprising a novel recombinant virus which replicates selectively in cells or tissues that are hypoxic or have an activated HIF pathway. The novel compositions of the invention comprise a recombinant virus genetically engineered to have a hypoxia/HIF-responsive element, or a multiplicity of such elements, operably linked to a promoter which is in turn operably linked to a nucleic acid(s) encoding a peptide(s) which regulates or modulates replication of the virus and/or encode a therapeutic molecule. The invention also includes constructs useful for screening of agents which interact with proteins or genes in the hypoxia-inducible pathway or are jointly translated under hypoxia and animal models useful for monitoring a variety of hypoxic conditions in a non-invasive manner.

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: A circular DNA molecule, useful for gene therapy, comprising at least one nucleic acid sequence of interest, characterised in that the region allowing the replication thereof has an origin of replication with a functionality in a host cell that requires the presence of at least one specific protein foreign to said host cell. A method for preparing same, cells incorporating said DNA molecules and uses thereof in gene therapy are also described.

Abstract: A lyophilized polynucleotide composition contains at least one polynucleotide and at least one cryoprotectant, wherein the ratio of the polynucleotide to cryoprotectant is from about 0.001 to about 1.0 part by weight polynucleotide per 1.0 part by weight of the cryoprotectant. This composition also contains from about 0.5 weight percent to about (6) weight percent water, based on the total weight of the final lyophilized polynucleotide composition. The polynucleotide composition of this invention is characterized by enhanced stability, in that it retains at least 90% supercoil over a time period of at least (10) days at a temperature of about 37° C. The lyophilized polynucleotide composition also has improved solubility. An improved process for lyophilization of polynucleotides employs a specific primary drying cycle, that results in the above-described stable, lyophilized polynucleotide composition.