Abstract: The invention concerns the use of a type II NAD(P)H dehydrogenase (NDH-II), or of a polynucleotide encoding said NDH-II, to increase the capacity of a green algae to produce hydrogen. Said polynucleotide is in particular useful for transforming said green algae, so as to improve its hydrogen production.

Abstract: A method of efficiently expressing Plasmodium AMA-1 ectodomain or a functional part, derivative, and/or analogue thereof, in a eukaryotic expression system. Preferably, the Plasmodium AMA-1 ectodomain is Pf AMA-1 ectodomain. This protein may be expressed in yeast, such as Pichia pastoris. Efficient expression is possible using a method for producing mRNA encoding the Plasmodium AMA-1 ectodomain in a yeast cell, comprising providing the yeast cell with a nucleic acid encoding Plasmodium AMA-1 ectodomain, the nucleic acid being modified to utilize the yeast cell's codon usage. Preferably, at least one putative yeast polyadenylation consensus sequence in the nucleic acid has been modified. More preferably, also at least one site in the protein that is generally glycosylated by eukaryotic expression systems, has been removed.

Abstract: The present invention provides a dual expression vector, and methods for its use, for the expression and secretion of a full-length polypeptide of interest in eukaryotic cells, and a soluble domain or fragment of the polypeptide in bacteria. When expressed in bacteria, transcription from a bacterial promoter within a first intron and termination at the stop codon in a second intron results in expression of a fragment of the polypeptide, e.g., a Fab fragment, whereas in mammalian cells, splicing removes the bacterial regulatory sequences located in the two introns and generates the mammalian signal sequence, allowing expression of the full-length polypeptide, e.g., IgG heavy or light chain polypeptide. The dual expression vector system of the invention can be used to select and screen for new monoclonal antibodies, as well as to optimize monoclonal antibodies for binding to antigenic molecules of interest.

Abstract: The present invention relates to a method for simultaneously surface expressing a target protein using a cofactor and an enzyme regenerating the cofactor on the cell surface. According to the present invention, it is possible to provide a microorganism capable of simultaneously surface expressing a target protein using a cofactor to transform a biochemical material at a high efficiency and an enzyme generating the cofactor without adding an expensive cofactor in a large amount.

Abstract: Bacteria are manipulated to create desirable output traits using dominant negative alleles of mismatch repair proteins. Enhanced hypermutation is achieved by combination of mismatch repair deficiency and exogenously applied mutagens. Stable bacteria containing desirable output traits are obtained by restoring mismatch repair activity to the bacteria.

Abstract: An object of the present invention is to provide a transformation system for Labyrinthulomycota that allows the elucidation of biosynthetic mechanisms of lipids such as PUFA and carotenoids as well as for the construction of a high production system and the design and development of novel functional lipid molecules by the control of the mechanisms. The present invention provides a method for introducing a transgene into a cell of Labyrinthulomycota, which comprises introducing into a cell of Labyrinthulomycota a recombinant vector comprising a transgene and a nucleotide sequence which is homologous to a part of chromosomal DNA of Labyrinthulomycota and is capable of homologous recombination with the chromosomal DNA, and then inducing homologous recombination in this homologous nucleotide sequence.

Abstract: The invention provides methods for screening multimeric antibodies produced by mammalian cells to find those that exhibit a biological function. The methods can be used to screen large numbers of antibodies, which may be cell surface, secreted, or intracellular antibodies. Antibodies can be screened to find those that bind antigen more avidly or those that compete with a ligand that binds to the antigen for binding. Any biological function that can be tested in vitro can be used to screen the antibodies. Nucleic acids encoding the antibodies that exhibit the biological function can be obtained in a number of ways.

Abstract: The present invention provides a method for identifying the presence of a protein inhibitor of a target protein in a sample, comprising the steps of a) contacting said sample with a cell, wherein said cell contains i) an inducible lethal overactivity mutation in a gene affecting the target protein; and ii) a mutation in a second gene, wherein the activity of the target protein is essential to the cell and the mutation in the second gene functionally compensates for any reduction in the activity of the target protein; b) inducing the lethal overactivity mutation; and subsequently c) assessing protein inhibition by comparing the degree of survival of the cell in the presence and the absence of said sample. Also provided are cells for use in said method.

Abstract: A recombinant double stranded RNA (dsRNA) nucleocapsid useful for the expression of dsRNA expression cassettes encoding passenger genes, such as, but not restricted to, vaccine antigens, bioactive proteins, immunoregulatory proteins, antisense RNAs, and catalytic RNAs, replicates in bacterial hosts and includes a P8 protein shell and three dsRNA segments where one of the segments includes a cap independent translation enhancer (CITE) operationally linked to a passenger gene.

Abstract: The present invention concerns methods and compositions for the construction of a series of stable vectors for genomic library construction useful in Gram negative species. In certain embodiments, the vectors contain the pBBR1 replicon, capable of to stable replication in a broad range of Gram negative species. In various embodiments, the plasmid vectors may also contain bidirectional, rho-independent transcriptional terminators flanking the multiple cloning site, which allows for greater insert stability, and thus, greater genomic representation. Each vector may vary in its selection marker region, mobilization function, and promoter used to express insert sequences. These vectors are of use in the screening of highly representational genomic libraries in a broad variety of Gram negative species.

Abstract: Embodiments of the present invention provide methods and compositions for microorganisms having increased alcohol tolerance. In certain embodiments, methods for using such microorganisms, and methods for identifying gene or genetic regions responsible for increased alcohol tolerance are contemplated.

Abstract: The present disclosure provides methods, devices and kits that permit large numbers of target biomolecules to be detected simultaneously in samples originating from a multi-sample holder, such as a multi-well plate. One specific example method is a method of making multiple substantial replicas of a biomolecular content of a multi-well sample holder. Devices and kits for carrying out the described methods are also provided.

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: Methods are provided for producing highly purified compositions of nucleic acids by using a highly streamlined and readily automated process. The methods use static mixers for lysing cells and precipitating debris, followed by centrifugation and ion exchange chromatography. The process may include a purification step using tangential flow ultrafiltration. A scaleable process for producing pharmaceutical grade plasmid DNA, useful for gene therapy, is provided.

Abstract: The invention provides an isolated and purified DNA molecule comprising at least one DNA segment, a biologically active subunit or variant thereof, of a circular intermediate of adeno-associated virus, which DNA segment confers increased episomal stability, persistence or abundance of the isolated DNA molecule in a host cell. The invention also provides a composition comprising at least two adeno-associated virus vectors.

Abstract: The invention relates to means and methods for regulating gene expression and production of proteinaceous molecules. The invention provides a method for producing a proteinaceous molecule in a cell comprising selecting a cell for its suitability for producing the proteinaceous molecule, providing a nucleic acid encoding the proteinaceous molecule with a nucleic acid comprising a STAR (STabilizing Anti-Repression) sequence, expressing the resulting nucleic acid in the cell and collecting the proteinaceous molecule. Providing at least one STAR sequence to a nucleic acid encoding a proteinaceous molecule will enhance production (yield) of the proteinaceous molecule by a host cell, increase the proportion of host cells with acceptable expression levels, and/or increase stability of a gene expression level.

Abstract: Novel benzoate- or anthranilate-inducible promoters, and novel tandem promoters, and variants and improved mutants thereof, useful for commercial prokaryotic fermentation systems, nucleic acid constructs containing the promoters, expression systems using them, methods for expressing proteins by use thereof, and proteins expressed thereby.

Abstract: The present invention relates to an animal feed made of an amount of cereal grain and a peptide or polypeptide expressed by a transformed organism, whereby the transformed organism can be included in the animal feed composition. The present invention also relates to a method for forming the animal feed wherein the method includes forming a transformed organism by transforming a yeast cell by inserting a nucleic acid molecule which will express a polypeptide desired for use in the animal feed.

Abstract: Disclosed herein are a method of preparing an L-methionine production strain by overexpressing proteins involved in L-methionine biosynthesis in an L-threonine production strain, a strain prepared by the method, and a method of producing L-methionine using the strain.

Abstract: The present invention is a group of cloning vector plasmids for use in constructing DNA molecules, such as transgenes, for the purpose of gene expression or analysis of gene expression. The present invention is also a method for using the cloning vector plasmids in a variable series of cloning steps to produce a final transgene product. The plasmid cloning vectors are engineered to minimize the amount of manipulation of DNA fragment components by the end user of the vectors and the methods for their use. Transgenes produced using the invention may be used in a single organism, or in a variety of organisms including bacteria, yeast, mice, and other eukaryotes with little or no further modification.