Abstract: The present invention includes perfusion bioreactors, automated cell culture systems, and methods for production of cells and cell-derived products.

Abstract: The subject matter of the present invention is a novel vector and the use thereof for producing a heterologous protein or a gene of interest, that can be used, for example, in the context of an immunization or gene therapy programme and concerns in particular a self-replicating vector lacking an antibiotic-resistance gene, comprising a sequence encoding the ccdA protein functionally linked to a first promoter, the sequence of the Cer locus and a heterologous sequence, functionally linked to a second promoter.

Abstract: The present invention provides a herpes virus which lacks a functional ICP34.5 encoding gene and which comprises two or more of—(i) a gene encoding a prodrug converting enzyme; (ii) a gene encoding a protein capable of causing cell to cell fusion; and (iii) a gene encoding an immunomodulatory protein.

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: Methods are provided for producing a vector that includes at least one splicable intron. In the subject methods, intron containing vectors are produced from donor and acceptor vectors that each include a site specific recombinase site, where the subject donor and acceptor vectors further include splice donor and acceptor sites that, upon site specific recombination of the donor and acceptor vectors, define an intron in the product vector of the recombination step. Also provided are compositions for use in practicing the subject methods, including the donor and acceptor vectors themselves, as well as systems and kits that include the same. The subject invention finds use in a variety of different applications, including the production of expression vectors that encode C-terminal tagged fusion proteins, the production of expression vectors that encode pure protein and not a fusion thereof, and the like.

Abstract: The present invention relates to a (first) method for producing inducible and/or repressible expression active linear RNA interference constructs comprising a PCR amplification of a source polynucleotide comprising the inhibitory RNA coding sequence of interest or comprising a PCR amplification of a DNA source comprising a promoter using a reverse primer comprising the inhibitory RNA coding sequence of interest. The present invention furthermore relates to a (second) method for producing inducible and/or repressible expression active linear gene constructs comprising a PCR amplification of a source expression polynucleotide comprising a promoter sequence and the DNA sequence of interest or comprising a PCR amplification using the DNA sequence as a template.

Abstract: Disclosed is a method for the mass production of a monomeric or dimeric immunoglobulin Fc region, free of initial methionine residues, using a recombinant expression vector comprising a nucleotide sequence coding for a recombinant immunoglobulin Fc region comprising an immunoglobulin Fc region linked at the N-terminus thereof to an immunoglobulin Fc region via a peptide bond.

Abstract: The invention relates to a mutated cell such as a bacteria or yeast in which the thyA gene coding thymidylate synthase includes a nonsense codon, preferably the amber codon, said nonsense codon replacing a codon coding an amino acid and inducing the interruption of thyA gene translation and the auxotrophy of the cell for the thymidine. Advantageously, the endA gene coding the endonuclease 1 and/or the recA gene coding the recombinase is inactivated in said mutated cell. The invention also relates to an expression plasmid including a transgene and a sequence of a suppressing ARM structural gene containing an anticodon that can be paired with the nonsense codon of the thyA gene and is specific of an amino acid capable of restoring the translation of the mutated thyA gene and thereby obtaining a protein of the wild or mutated type having a thymidylate synthase activity. The invention also relates to a method for the multiplication of the expression plasmid.

Abstract: The invention relates to compositions, specifically novel nucleic acid constructs encoding a cardiovirus 2A polypeptide operably linked to suitable promoters. Also, disclosed are methods whereby the nucleic acid constructs are introduced into cells or cell free systems to regulate cellular mRNA transcription and cap-dependent or internal ribosomal entry site (IRES)-dependent mRNA translation.

Abstract: The present invention provides a reagent for cancer cell detection or cancer diagnosis. The present invention relates to a reagent for cancer cell detection, comprising a recombinant virus where a replication cassette comprising a promoter from human telomerase, an E1A gene, an IRES sequence and an E1B gene in this order is integrated in E1 region of the viral genome and a labeling cassette comprising a gene encoding a labeling protein and a promoter capable of regulating the expression of the gene encoding the labeling protein is integrated in E3 region of the viral genome.

Abstract: The various embodiments of the present invention relate generally to plasmid DNA preparations and to methods for producing and using such preparations.

Abstract: The present invention provides methods of producing dsDNA molecules that can be used to mediate RNA interference (RNAi). These methods include the production of hairpin DNAs including random sequences, and the use of convergent promoters to co-express sense and antisense RNAs. As such, the invention allows the production of random short hairpin RNA (shRNA) and a small interfering RNA (siRNA) expression libraries for forward genetic screening.

Abstract: The invention provides rhinovirus vectors, which can be used in the delivery of immunogens, such as influenza virus immunogens, and corresponding compositions and methods.

Abstract: The present invention relates to plant cells and plants, which are genetically modified, wherein the genetic modification leads to the reduction of the activity of a starch phosphorylating OK1 protein in comparison with corresponding wild type plant cells or wild type plants that have not been genetically modified. Furthermore, the present invention relates to means and methods for the manufacture of such plant cells and plants. Plant cells and plants of this type synthesise a modified starch. The present invention therefore also relates to the starch synthesised by the plant cells and plants according to the invention, methods for the manufacture of this starch, and the manufacture of starch derivatives of this modified starch, as well as flours containing starches according to the invention. Furthermore, the present invention relates to nucleic acid molecules, which are suitable for manufacturing plants according to the invention.

Abstract: The present invention is directed to a replicative, thermostable plasmid. In particular, the present invention is directed to a replicative, thermostable plasmid comprising a sequence derived from the pB6A plasmid and at least one functional unit comprising a sequence that is not found in plasmid pB6A.

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 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: 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: A baculovirus expression vector achieves dual functions of (1) subunit vaccine by displaying the influenza surface protein for humoral immune responses; and (2) DNA vaccine by expressing influenza surface protein for long-acting cellular immune response. A method for inducing immunogenicity in a host is also disclosed.

Abstract: The invention features new helper virus-free methods for making herpesvirus amplicon particles that can be used in immunotherapies, including those for treating any number of infectious diseases and cancers (including chronic lymphocytic leukemia, other cancers in which blood cells become malignant, lymphomas (e.g. Hodgkin's lymphoma or non-Hodgkin's type lymphomas). Described herein are methods of making helper virus-free HSV amplicon particles; cells that contain those particles (e.g., packaging cell lines or patients' cells, infected in vivo or ex vivo); particles produced according to those methods; and methods of treating a patient with an hf-HSV particle made according to those methods.

Abstract: A novel human collagen-like protein CLAC occurring in brain amyloid and its precursor CLAC-P; genes encoding the same; cDNA of mouse CLAC-P and its deduced amino acid sequence; antibodies specific to these proteins; and methods of diagnosing treating and preventing Alzheimer's disease by using the same.

Abstract: Provided herein are Salmonella enteritidis 13A strains and compositions comprising these strains. Also provided are methods of enhancing an immune response against Influenza A and methods of reducing morbidity associated with an Influenza A infection. Methods of enhancing an immune response to a vaccine vector by expressing a polypeptide of CD 154 capable of binding CD40 are also disclosed. Methods of developing a bacterial vaccine vector are disclosed. Methods of generating scarless site-specific mutations in a bacterium are also disclosed.

Abstract: A method for the recombination of a gene is disclosed. The method involves the design of unpaired forward and reverse primers having homology to the 5? end of one template and to the 3? end of another template. Short primer extension periods results in a recombined template having paired 5? and 3? ends that can then be amplified. The amplified sample is devoid of any parental template.

Abstract: The present invention provides a method of covalently joining a DNA strand to an RNA strand comprising (a) forming a topoisomerase-DNA intermediate by incubating a DNA cleavage substrate comprising a topoisomerase cleavage site with a topoisomerase specific for that site, wherein the topoisomerase-DNA intermediate has one or more 5? single-strand tails; and (b) adding to the topoisomerase-DNA intermediate an acceptor RNA strand complementary to the 5? single-strand tail under conditions permitting a ligation of the covalently bound DNA strand of the topoisomerase-DNA intermediate to the RNA acceptor strand and dissociation of the topoisomerase, thereby covalently joining the DNA strand to the RNA strand. The present invention also provides a method of tagging a 5? end of an RNA molecule. The present invention further provides a DNA-RNA molecule which has been joined in vitro by the use of a topoisomerase. The present invention also provides a method of tagging a 5? end of an mRNA.

Abstract: A computer implemented method for generating nucleotide sequences containing candidate homing endonuclease genes (HEGs). A search is performed in a database stored on a storage medium of nucleotide sequences for amino acid sequences having a subsequence having a homology level with the translation of a subsequence of one or more predetermined HEGs. For each amino acid sequence generated by the search, one or more nucleotide sequences are retrieved encoding the amino acid sequence. The results of this search used in a second search of a database stored on a storage medium to generate the HEG containing sequences.

Abstract: The present invention relates to cloning target nucleic acids using phage packaging mechanisms. Packaging initiation sites may be introduced into the target DNA. Components of a phage packaging system may be combined with the target DNA to package the DNA into phage capsids. The packaged DNA may be used to create a library of target nucleic acids, or it may be sequenced.

Abstract: JNK-interacting protein 1 (JIP-1), an inhibitor of the JNK1 protein, and methods of treating a pathological condition or of preventing the occurrence of a pathological condition in a patient by the administration of a therapeutically effective amount of JIP-1 polypeptides, peptides, peptide mimetics, or nucleic acids are described.

Abstract: In the absence of substantial sequence overlap between a recombinant adenoviral vector and the genome of a packaging cell, helper-dependent E1-containing particles (HDEP) can be formed at low frequency. The invention provides means and methods reducing or preventing the generation of HDEP. To this purpose, novel packaging cells and methods of making these are provided.

Abstract: The invention provides anti-OX40L antibodies, and compositions comprising and methods of using these antibodies.

Abstract: A recombinant vector for delivering A3G genes into human cells comprising (i) a gene expression block including an A3G gene selected from a wild type A3G gene represented by SEQ ID NO: 1 and a mutant A3G gene and (ii) a group of elements from a modified lentiviral vector including lentiviral regions of packaging signal (?, psi), LTRs, RRE, and PBS; wherein said A3G gene is operably linked to the packaging signal (?, psi), LTRs, RRE, and PBS.

Abstract: A viral vector production system is provided which system comprises: (i) a viral genome comprising at least one first nucleotide sequence encoding a gene product capable of binding to and effecting the cleavage, directly or indirectly, of a second nucleotide sequence, or transcription product thereof, encoding a viral polypeptide required for the assembly of viral particles; (ii) a third nucleotide sequence encoding said viral polypeptide required for the assembly of the viral genome into viral particles, which third nucleotide sequence has a different nucleotide sequence to the second nucleotide sequence such that said third nucleotide sequence, or transcription product thereof, is resistant to cleavage directed by said gene product. The viral vector production system may be used to produce viral particles for use in treating or preventing viral infection.

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: The present invention relates to methods of screening for expression of a soluble candidate protein within an expression library of candidate proteins. The method involves fusing each candidate protein in the library to a peptide substrate and identifying cells that express soluble candidate protein by detecting enzymatic modification of the peptide substrate.

Abstract: The present invention provides: a novel DNA, a carcinoma-associated gene comprising the DNA, a recombinant protein encoded by the DNA, an antibody binding to the protein, an anti-carcinoma agent comprising the antibody, a low-molecular-weight compound binding to the protein, and a screening system. An example of such a novel DNA is a DNA comprising a nucleotide sequence encoding the following polypeptide (a) or (b): (a) a polypeptide, consisting of an amino acid sequence identical to or substantially identical to the amino acid sequence represented by SEQ ID NO: 2; or (b) a polypeptide, consisting of an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 2 by deletion, substitution, or addition of one or a plurality of amino acids and having biological activity substantially equivalent to the functions of the polypeptide (a).

Abstract: The present invention relates to a method for producing a protein, novel nucleic acid fragment, an expression vector, and a method for enhancing the protein production in Pichia. The present invention provides that the combination of the modified signal peptide, increased copy number of the gene, and/or overexpressed Sec4p resulted in high level secretion of proteins.

Abstract: This invention provides a method for combining overlapping DNA molecules comprising: (a) providing first and second DNA fragments, the first having a region homologous to a region in the second; (b) tagging the first DNA fragment with a selectable marker; (c) cloning the first DNA sequence into a retrieval vector to form a DNA-vector complex; (d) linearizing the DNA-vector complex; and (e) inserting the first DNA fragment from the DNA-vector complex into the second DNA fragment using homologous recombination to form a combined DNA molecule; and (f) removing the selectable marker, thereby generating a combined DNA molecule. The invention further provides a vector for retrieving and inserting a selected DNA molecule into a target DNA molecule.

Abstract: The present inventors succeeded in producing non-replicating SeV vectors whose genomic RNAs lack all genes for the NP, P, and L proteins, which are RNP-constituting proteins. The present inventors confirmed that the NP/P/L-deficient SeV vectors carrying a marker gene such as GFP provide high productivity, and high transfer and expression efficiencies of foreign genes (high MOI infection is essential for achieving high expression levels). By lacking the L gene or two or more of the NP, P, and L genes, the vectors of the present invention enable lowering the level of virus-derived proteins expressed in host cells, thereby reducing the immunogenicity upon in vivo administration.

Abstract: The invention relates to a pORT plasmid containing a sequence encoding all or part of a disintegrin domain of metargidin (RDD) or a derivative thereof under the control of strong cytomegalovirus promoter, in particular a plasmid having the sequence shown in SEQ ID NO:2.

Abstract: The invention provides viral vectors, such as chimeric flavivirus vectors, including foreign peptides inserted into the target proteins of the vectors, methods of making and using these vectors, and compositions including the vectors.

Abstract: General methods and strains of bacteria are described, that dramatically simplify and streamline plasmid DNA production. In one preferred embodiment, endolysin mediated plasmid extraction combined with flocculation mediated removal of cell debris and host nucleic acids achieves increased yield and purity with simplified downstream purification and reduced waste streams, thus reducing production costs.

Abstract: The present invention relates to the production of covalently closed circular (ccc) recombinant DNA molecules such as plasmids, cosmids, bacterial artificial chromosomes (BACs), bacteriophages, viral vectors and hybrids thereof, and more particularly to vector modifications that improve production yield of said DNA molecules in fermentation culture.

Abstract: An obligate heterodimer meganuclease consisting of a first and a second monomer, deriving from two different homodimeric LAGLIDADG (SEQ ID NO: 66) endonuclease monomers (parent monomers), and having at least one pair of mutations interesting corresponding residues of said parent monomers which make an intermolecular interaction between the two monomers of each parent homodimeric LAGLIDADG (SEQ ID NO: 66) endonuclease, a vector encoding said meganuclease, a cell, an animal or a plant modified by said vector and the use of said meganuclease and derived products for molecular biology, genome engineering and genome therapy.

Abstract: The present invention relates generally to a novel plant promoter. More particularly, the present invention provides a plant promoter capable of induction by physical and/or environmental stimuli in cells in which the promoter is indigenous and, in the absence of any negative regulatory mechanism, is capable of constitutive expression in cells in which the promoter is non-indigenous. The present invention is further directed to derivatives of the subject promoter including modular forms of the promoter which are, for example, inducible by different physical and environmental stimuli or which are constitutively expressed. The promoter of the present invention has a range of uses including directing expression of genes conferring useful traits on plants.

Abstract: Recombinant lentiviruses and transfer vectors for transgene delivery as well as methods for gene therapy using such vectors are disclosed. The invention provides a third generation lentiviral packaging system and a set of vectors for producing recombinant lentiviruses, as well as novel tissue specific enhancer and promoter elements useful for optimizing liver specific transgene delivery. The transgene is preferably a blood clotting factor such as human factor IX (hFIX) or human factor VIII (hFVIII) and can be used for treatment of hemophilia.

Abstract: The present invention relates to a method for producing a DNA chip, which comprises the steps of: (a) cloning a probe, where a linker is coupled to one or both ends of an oligonucleotide to be integrated on a slide, into a vector; (b) transforming host cells with the vector; (c) culturing the transformed host cells, to recover the probe where the linker is coupled to one or both ends of the oligonucleotides; and (d) integrating the recovered double-helical probes on a slide. Also, the present invention relates to a DNA chip for HPV diagnosis produced by the method, and a method for diagnosing the presence or genotype of HPV using the DNA chip.

Abstract: The present invention relates to compositions comprising a therapeutically effective amount of genetically modified cells containing a genetic construct expressing a TGF? inhibitor effective to reduce expression of TGF?, where the genetically modified cells are non-small cell lung cancer (NSCLC) cells or small cell lung cancer (SCLC) cells, and related methods.

Abstract: The present invention relates generally to the field of molecular biology and genomics. More specifically, the present invention concerns the cloning of nucleic acid molecules and the production of nucleic acid libraries, as well as the expression of recombinant proteins and bactofection.

Abstract: The present invention relates generally to nucleic acid constructs, which are useful for inserting a nucleotide sequence of interest into a target nucleic acid molecule via homologous recombination. The present invention also relates to methods for producing such constructs.

Abstract: The present invention provides compositions and methods for rapid assembly of one or more assembled polynucleotides from a plurality of component polynucleotides. The methods of the invention utilize circular nucleic acid vectors that comprise a DNA segment D flanked by an annealable linker sequence, annealable linker sequence pairs LA and LB, or annealable linker sequence/primer binding segment pairs LA and PB or PA and LB. Restriction endonuclease digestion of a plurality of vectors containing the DNA segments to be assembled generates a plurality of DNA fragments comprising the elements PA-D-LB, LA-D-LB, and LA-D-PB or D-LB, LA-D-LB, and LA-D. The sequences of annealable linker sequences LA and LB provide complementary termini to the DNA fragments, which are utilized in host cell mediated homologous recombination or together with promer binding segments PA and PB in a polymerase cycling assembly reaction for the ordered assembly of the various DNA segments into one or more assembled polynucleotides.

Abstract: The present invention relates to a deoxyribonucleic acid (DNA) comprising at least one promoter sequence, which is derived from a wild-type promoter of a methyltrophic yeast, whose transcription efficiency is modulated in comparison to the efficiency of the wild-type promoter by inserting or modifying a DNA binding site. The invention also relates to host cells, expression vectors, kits and methods for producing proteins while using the inventive DNA, as well as to different uses of the same and to a method for producing expression vectors.