Abstract: Improved methods and prokaryotic expression systems for producing recombinant proteins using Sec-system secretion signal peptides are described using Pseudomonas fluorescens Sec secretion systems. Specific novel Sec-system secretion signal peptides are described, as are fusion proteins and coding sequences for improved secretion of recombinant proteins and peptides.

Abstract: The present invention relates to novel methods for sequencing and mapping genetic markers in polynucleotide sequences using Type-IIs restriction endonucleases. The methods herein described result in the “capturing” and determination of specific oligonucleotide sequences located adjacent to Type-IIs restriction sites. The resulting sequences are useful as effective markers for use in genetic mapping, screening and manipulation.

Abstract: The present invention describes a single-protein production (SPP) system in living E. coli cells that exploits the unique properties of an mRNA interferase, for example, MazF, a bacterial toxin that is a single stranded RNA- and ACA-specific endoribonuclease, which efficiently and selectively degrades all cellular mRNAs in vivo, resulting in a precipitous drop in total protein synthesis. Concomitant expression of MazF and a target gene engineered to encode an ACA-less mRNA results in sustained and high-level (up to 90%) target expression in the virtual absence of background cellular protein synthesis. Remarkably, target synthesis continues for at least 4 days, indicating that cells retain transcriptional and translational competence despite their growth arrest. SPP technology works well for yeast and human proteins, even a bacterial integral membrane protein.

Abstract: L-lysine-producing strains of corynebacteria with enhanced lysE gene (lysine export carrier gene), in which strains additional genes chosen from the group comprising the dapA gene (dihydrodipicolinate synthase gene), the lysC gene (aspartate kinase gene), the dapB gene (dihydrodipicolinate reductase gene) and the pyc gene, but especially the dapA gene and the lysC gene (aspartate kinase gene), are enhanced and, in particular, over-expressed, and a process for the preparation of L-lysine.

Abstract: A molecule transfer and delivery system includes a loading zone (50) for loading a targeted cargo molecule (30) on a carrier molecule (20) making use of a hybridization nature of nucleotides, and an unloading zone (40) for unloading the targeted cargo molecule from the carrier molecule making use of the strand exchange nature of the nucleotides. Towing nucleotide chains including the first single-stranded nucleotides (21) with the first length are bound to the carrier molecule, and the second single-stranded nucleotides (31) with the second length longer than the first length are bound to the targeted cargo molecule. The third single-stranded nucleotides (41) with the same length as the second single-stranded nucleotides are bound to the unloading zone.

Abstract: The invention relates to a method for the transfer of molecular substances, for example proteins or nucleic acids in cells, in the case of using DNA combined with a possible gene expression. A prokaryotic nucleic acid-binding protein is used for the transfer, which is preferably obtained from a thermostable organism. Where the substance to be transferred is a nucleic acid, the protein forms a reversible complex with the nucleic acid. The prokaryotic protein condenses and compacts the nucleic acids. Said nucleic acids can be taken up in the target cells after suitable incubation.

Abstract: The present invention relates to a method for determining the ideal time for and outcome of reproductive health procedures including in vitro fertilization by establishing a correlation between the successful outcome of said procedure and the spectra of a body fluid obtained using a chosen analytical modality for a population of patients, acquiring for a patient a spectrum of the body fluid of the patient using said chosen modality.

Abstract: A novel nuclear receptor, termed the steroid and xenobiotic receptor (SXR), a broad-specificity sensing receptor that is a novel branch of the nuclear receptor superfamily, has been discovered. SXR forms a heterodimer with RXR that can bind to and induce transcription from response elements present in steroid-inducible cytochrome P450 genes in response to hundreds of natural and synthetic compounds with biological activity, including therapeutic steroids as well as dietary steroids and lipids. Instead of hundreds of receptors, one for each inducing compound, the invention SXR receptors monitor aggregate levels of inducers to trigger production of metabolizing enzymes in a coordinated metabolic pathway. Agonists and antagonists of SXR are administered to subjects to achieve a variety of therapeutic goals dependent upon modulating metabolism of one or more endogenous steroids or xenobiotics to establish homeostasis.

Abstract: The invention concerns making and evaluating synthetic regulatory regions for controlling gene expression. The invention features a method for identifying transcription factor binding sites and a method for evaluating the regulatory functions of synthetic regulatory regions.

Abstract: Methods of screening for compounds which are, for example, capable of modulating amino acid-DNA interaction, modulating DNA replication, modulating cell proliferation, and for identifying compounds which inhibit cellular proliferation caused by cancer, are provided.

Abstract: Described is the production of proteins in a host cell. Even more specifically, described are methods for improving expression of two or more proteins in a cells or host cell. The methods are suited for production of, for example, recombinant antibodies that can be used in pharmaceutical preparations or as diagnostic tools. In certain embodiments, provided are methods for obtaining a cell that expresses two or more proteins comprising providing the cell with two or more protein expression units encoding two or more proteins, characterized in that at least two of the protein expression units comprise at least one STAR sequence.

Abstract: The present invention relates to a method for generating a novel form of life comprising the steps consisting of: a) irreversible alteration of the genome of a microbial clone; b) cultivation of a vast population of microbial cells originating from the altered clone obtained in step a) during numerous generations under conditions allowing selection for a higher and stable proliferation rate; c) isolation of descendant clones within the cultivated population of step b) still bearing the alteration of step a).

Abstract: Disclosed is a method for producing a biological organic material in a liquid droplet comprising a culture medium composed of specified ingredient. The method comprises the steps of: providing a substantially spherical liquid droplet on a substrate in an unadhered state, the substrate preferably having such a water-repellent surface that the water contact angle becomes 150° or greater; and culturing a biological organic material in the liquid droplet.

Abstract: Improvements in plasmid DNA production technology are needed to insure the economic feasibility of future DNA vaccines and DNA therapeutics. General methods are described, by means of which it is possible to dramatically increase plasmid DNA productivity in a fermentor. These processes feature Fed-batch fermentation strategies, combined with novel growth and induction phase temperature shifts.

Abstract: The invention concerns a kit for detecting and/or quantifying a ligant (C), present in an analysis sample, by a receptor (A), characterized in that it comprises the following reagents: a single-stranded or double-stranded nucleotide sequence (B), and said receptor (A) consisting of a monomeric or multimeric proteinic entity including at least a first recognition site specific for said ligand (C) and a second recognition site specific for said nucleotide sequence (B); said ligand (C) being capable of positively or negatively modulating fixing of the receptor (A) by its specific recognition site on the nucleotide sequence (B).

Abstract: A nanostructured molecular delivery vehicle comprising magnetic materials and configured to receive passenger biomolecules. The application of a an appropriate magnetic field having a gradient orients and drives the vehicle into a biological target, which may comprise cells, cell masses, tissue slices, tissues, etc. Under the control of the magnetic field, these vehicles can penetrate cell membranes. Then, the biomolecules carried by the vehicle can be released into the cells to perform their functions. Using this “nanospearing” technique, unprecendented high transfection efficiency has been achieved in several difficult-to-transfect cells. These include, but are not limited to, Bal 17 cells, ex vivo B cells, primary cultured cortical neurons, etc. This method advances the state of the art, providing an improved technique for the introduction of exogenous molecules to cells, with the clinical applications including, but not being limited to, drug delivery, gene therapy, vaccination, etc.

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

Abstract: Improved vector constructs useful in the expression of double-stranded RNA are provided. The constructs are particularly useful for expression of double-stranded RNA in vitro and in vivo.

Abstract: There is provided a method by which multiple types of substances desired to be transferred into cells can be continuously transferred into multiple types of cells by a convenient procedure, a cell in which the substance desired to be transferred into cells has been taken up by this method, and an apparatus for transferring a substance into cells by this method. The foregoing objects can be achieved by electrospraying cells with a liquid free from the substance to be transferred into cells while the cells are kept in contact with the substance to be transferred into cells, or first electrospraying cells with a liquid free from the substance to be transferred into cells and then bringing the cells into contact with the substance to be transferred into cells.

Abstract: The present invention provides a method and apparatus for transferring an agent into a cell. The method includes the steps of providing an agent outside of a cell and generating a vapor bubble and a plasma discharge between an avalanche electrode and a conductive fluid surrounding the cell. The vapor bubble and plasma discharge generate a mechanical stress wave and an electric field, respectively. The combination of this mechanical stress wave and electric field results in permeabilization of the cell, which in turn results in transfer of the agent into the cell.