Abstract: The present invention relates to the production and use of covalently closed circular (ccc) recombinant plasmids, and more particularly to vector modifications that improve expression of said DNA molecules in the target organism.

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: Improved DNA vaccine plasmids are disclosed. The improved plasmids eliminate all extraneous sequences, and have superior eukaryotic expression, improved yield and stability during bacterial production, facilitate flexible targeting of antigens to various intracellular destinations, and novel RNA-based functionality. These vectors are utilized in novel immunization methods wherein combinations of immunostimulatory DNA vaccine plasmids that target antigens to various intracellular destinations are used to elicit improved immune response.

Abstract: The present invention relates to the propagation of covalently closed circular recombinant DNA molecules such as plasmids, cosmids, bacterial artificial chromosomes (BACs), bacteriophages, viral vectors and hybrids thereof, and more particularly is strain modifications that improve strain viability, plasmid stability, plasmid production yield, and plasmid-directed protein production yield, using said DNA molecules in fermentation culture.

Abstract: The present invention relates to recombinant DNA molecules such as plasmids, non-viral vectors, viral vectors and hybrids thereof, and more particularly to vector modifications that improve expression of said DNA molecules in cell lines and organisms.

Abstract: Improved DNA vaccine plasmids are disclosed that contain novel immunostimulatory RNA compositions. The improved plasmids eliminate all extraneous sequences, incorporate a novel antibiotic free short RNA based selectable marker, increase eukaryotic expression using a novel chimeric promoter, improve yield and stability during bacterial production, and improve immunostimulation. These vectors are utilized in immunization to elicit improved immune responses or therapy to induce type 1 interferon production.

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: Improvements in strain engineering technology are needed to insure the economic feasibility of future engineered recombinant organisms for industrial biotechnology. Disclosed herein are rapid, efficient methods (Genome Mass Transfer) that facilitate introduction of new selectable traits into a target microbial host. In one preferred embodiment, methods for high efficiency electroporation mediated transfer of donor DNA into a recipient microbial cell are disclosed.

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: Improved recombinant retrotransposon vectors for gene transfer are disclosed. The synthetic vectors are truncated so as to reduce or altogether eliminate homologous recombination with retroviral helper sequences found in helper cells used to propagate the vectors, making them safer for use in humans and providing more space for therapeutic genes. The vectors transmit foreign DNA efficiently, are stable, enable abundant RNA expression from the retrotransposon transcriptional promoter, and through their diversity permit many useful applications in therapeutics and transgenics. Methods are described for rescuing tissue-specifics promoters obtaining expression in primary cells, mapping the genome and other techniques of therapeutic and transgenic utility.

Abstract: A chimeric viral packaging signal is described for the transmission of genetic materials via retrovirus. The packaging signal contains an essential packaging nucleic acid sequence and a non-essential nucleic acid sequence. The packaging signal lacks gag gene sequences, and has approximately one order of magnitude greater infectivity than retrovirus-derived packaging signals without gag gene sequences. The packaging signal of the invention can be used to transmit genetic material for gene therapy, cell therapy, or other biotechnological applications.

Abstract: Improved recombinant retrotransposon vectors for gene transfer are disclosed. The synthetic vectors are truncated so as to reduce or altogether eliminate homologous recombination with retroviral helper sequences found in helper cells used to propagate the vectors, making them safer for use in humans and providing more space for therapeutic genes. The vectors transmit foreign DNA efficiently, are stable, enable abundant RNA expression from the retrotransposon transcriptional promoter, and through their diversity permit many useful applications in therapeutics and transgenics. Methods are described for rescuing tissue-specifics promoters obtaining expression in primary cells, mapping the genome and other techniques of therapeutic and transgenic utility.

Abstract: Improved recombinant retrotransposon vectors for gene transfer are disclosed. The synthetic vectors are truncated so as to reduce or altogether eliminate homologous recombination with retroviral helper sequences found in helper cells used to propagate the vectors, making them safer for use in humans and providing more space for therpeutic genes. The vectors transmit foreign DNA efficiently, are stable, enable abundant RNA expression from the retrotransposon transcriptional promoter, and through their diversity permit many useful applications in therapeutics and transgenics. Methods are described for rescuing tissue-specific spromoters obtaining expression in primary cells, mapping the genome and other techniques of therapeutic and transgenic utility.

Abstract: A process for the transfer and/or expression of any gene into cells, organisms or species is disclosed. The process involves isolating the gene, introducing the gene into a vector comprised of at least one retrotransposon genetic element to provide a hybrid gene. The hybrid gene is introduced into a donor cell capable of packaging and transmitting the retrotransposon to other cells, organisms or species. The hybrid gene is transferred to a recipient cell wherein the hybrid gene replicates by reverse transcription and is inserted into the recipient cell's genome. The gene is preferably expressed as RNA and/or protein, giving the phenotype of the gene.