Abstract: The present invention is directed to the method and apparatus for the cytoplasmic loading of macromolecules into living cells by an impact-mediated procedure that impacts the cells with a predetermined number of solid particles in a blast of propellant gas. More specifically, the present invention is directed to an impact-mediated procedure that is altered by gravitational conditions and is preferably carried out under hypergravity conditions. Further, the present invention is directed to an IML method and apparatus for consistently and reproducibly loading macromolecules into the cytoplasm of living cells via membrane wounding at significantly higher efficiencies than can be accomplished using existing methodologies. The IML procedure directs a blast of propellant gas through a rupturable membrane on which solid particles are supported in order to achieve insertion of a predetermined number of particles into the propellant blast.

Abstract: The invention relates to the human herpesvirus type 6 protein p100 and parts thereof having its specific immunological properties. It further relates to antibodies directed to them and to the corresponding DNA sequences. They can be used in pharmaceutical or diagnostic compositions, optionally together with other HHV-6 proteins or the corresponding DNA sequences.

Abstract: This invention relates to DNA constructs for inserting heterologous gene sequences into a host genome so as to obtain expression of the heterologous gene, to methods of inserting heterologous gene sequences into a host genome, and to organisms carrying modified host genomes. Specifically, the DNA constructs of this invention contain an expression unit of an internal ribosome binding site (IRES) coupled to a heterologous gene sequence. This expression unit is flanked at the 5' and 3' ends by DNA sequences that enable homologous recombination or integration of the construct with the DNA of a targeted host to obtain expression of the heterologous gene in the host.

Abstract: A pair of degenerate oligonucleotide primers can amplify transglutaminase-specific fragments of known transglutaminase genes. The primers are also used to obtain new transglutaminase gene products. The nucleotide sequence of a novel transglutaminase gene (termed TG.sub.X) is presented.

Abstract: Determining orientation and direction of DNA sequences. A method by which fluorescence in situ hybridization can be made strand specific is described. Cell cultures are grown in a medium containing a halogenated nucleotide. The analog is partially incorporated in one DNA strand of each chromatid. This substitution takes place in opposite strands of the two sister chromatids. After staining with the fluorescent DNA-binding dye Hoechst 33258, cells are exposed to long-wavelength ultraviolet light which results in numerous strand nicks. These nicks enable the substituted strand to be denatured and solubilized by heat, treatment with high or low pH aqueous solutions, or by immersing the strands in 2.times.SSC (0.3M NaCl+0.03M sodium citrate), to name three procedures. It is unnecessary to enzymatically digest the strands using Exo III or another exonuclease in order to excise and solubilize nucleotides starting at the sites of the nicks.

Abstract: The present invention relates, in general, to methods of stimulating phagocytosis and thereby combating infection and/or modulating immune complex disease, in particular, to methods of modulating the number and type of Fc receptors present on cells that normally possess such receptors, including monocytes and macrophages, as well as on cells that normally do not possess Fc receptors, such as fibroblasts, and to compounds and compositions suitable for use in such methods.

Abstract: Disclosed are substantially pure nucleic acid sequences encoding vertebrate cdc37. The protein encoded by this nucleic acid is characterized by the ability to bind specifically to mammalian proteins selected from the group consisting of cdk4, cyclin D, cdk2, Rb, hsp90, hsp60, hsp70, Raf-1, MEK-1, SAPK, MAPK, ERK1, P13-K and the src family of tyrosine kinases. In addition, the protein is further characterized by the ability to bind glycosaminoglycan. Also disclosed are diagnostic and therapeutic applications based on the isolation of the nucleic acid sequences.

Abstract: NIa protease genes of papaya ringspot virus strains FLA83 and USA P-type (HA-attenuated strain) are provided.

Abstract: The present invention describes methods of producing milligram quantities of three forms of purified Stat1 protein from recombinant DNA constructs. In addition, the Stat proteins may be isolated in their phosphorylated or nonphosphorylated forms (Tyr 701). The proteins can be produced in baculovirus infected insect cells, or E. coli. A compact domain in the amino terminus of Stat1.alpha. was isolated and found to enhance DNA binding due to its ability to interact with a neighboring Stat protein. A relatively protease-resistant recombinant truncated form of the Stat protein was isolated in 40-50 mg quantities. Purification of the Stat proteins were performed after modifying specific cysteine residues of the Stat proteins to prevent aggregation. Activated EGF-receptor partially purified from membranes by immunoprecipitation was shown to be capable of in vitro catalysis of the phosphorylation of the tyrosine residue of Stat1 known to be phosphorylated in vivo.

Abstract: A process is provided for the bioconversion of a carbon substrate to 1,3-propanediol by a single organism utilizing either microorganisms containing the genes encoding for an active glycerol or diol dehydratase enzyme by contacting these organisms with a carbon substrate under the appropriate fermentation conditions.

Abstract: Method of modifying DNA by subjecting the DNA to a mutation-inducing treatment. The method includes the steps of bringing the DNA to be mutated and a gene encoding a mutation-inducing non-DNA polymerase protein together in cells, growing the cells in the presence of a stress factor, and selecting the mutant cells which have developed a desirable trait in the presence of the stress factor.

Abstract: The present invention relates to a method by which one can target an undesired target molecule or target antigen, preferably an endogenous protein. The method comprises the intracellular expression of an antibody capable of binding to the target. A DNA sequence is delivered to a cell, the DNA sequence contains a sufficient number of nucleotides coding for the portion of an antibody capable of binding to the target operably linked to a promoter that will permit expression of the antibody in the cell(s) of interest. The antibody is then expressed intracellularly and binds to the target, thereby disrupting the target from its normal actions.

Abstract: The present invention provides liver parenchymal cells having a clonal growth ability, which possesses at least one of the cell biological properties such as: presence of peroxysome; being positive to hepatocyte-markers; being partially positive to neoplastic hepatocyte-markers or immature hepatocyte-markers; being positive to antibodies against the surface antigens of ovall cells; and being partially positive to bile duct cell-markers. The present invention also provides a method for obtaining such cells and a method for subculturing such cells.With the liver parenchymal cells above, it will be possible to research in detail the process of development and differentiation, the mechanisms of growth and functional expression of hepatic cells, and to open up a new way to clalification of mechanisms or hepatoma and various other diseases and to development of therapeutic method against these disseases.

Abstract: Accessory functions capable of supporting efficient recombinant AAV (rAAV) virion production in a suitable host cell are provided. The accessory functions are in the form of one or more vectors that are capable of being transferred between cells. Methods of producing rAAV virions are also provided. The methods can be practiced to produce commercially significant levels of rAAV particles without also generating significant levels of infectious helper virus or other contaminating by-products.

Abstract: The present invention provides a composition of matter, comprising: DNA encoding a viral Vpx protein fused to DNA encoding a virus inhibitory protein. In another embodiment of the present invention, there is provided a composition of matter, comprising: DNA encoding a viral Vpr protein fused to DNA encoding a virus inhibitory protein. Also provided are various methods of delivering a virus inhibitory molecule to a target in an animal. Further provided is a pharmaceutical composition.

Abstract: Methods for screening for a drug that increases expression of the human apolioprotein (apo) AI gene and related DNA constructs. A first method has the steps of: (a) introducing into mammalian cells a DNA construct including, functionally joined together in the 5'.fwdarw.

Abstract: The invention relates to inhibitors suppressing the activity of transacting proteins which are essential for the general control of vertebrate MHC class II gene expression and methods for identifying the same. The invention additionally relates to pharmaceutical compositions containing said inhibitors, preferably for the treatment of diseases which are associated with an aberrant expression of MHC class II genes.

Abstract: Novel deletion mutants of the class II transactivator protein (CIITA) are disclosed. Isolated DNA encoding such mutants are also disclosed, as are recombinant DNA vectors containing said DNA, and antibodies that specifically bind the mutant CIITA proteins. Additionally, methods of treating disorders characterized by abnormal expression of major histocompatibility complex (MHC) antigens by administering novel mutants of the present invention are also disclosed.

Abstract: The present invention provides a system for controlled transgene transcription using chimeric activator and repressor proteins functioning in a novel regulatory network. The target transgene is actively silenced in non-inducing conditions by a novel class of chimeric proteins consisting of the DNA-binding tetracycline repressor fused to distinct multimerized eukaryotic transcriptional repression domains. In the presence of a tetracycline "inducer", the repressor does not bind to the promoters for both the target gene and for another regulatory gene encoding a transactivator (e.g., GAL4-VP16). Under these inducing conditions, the transactivator activates expression of the target transgene and of its own gene, in an additional autoregulatory positive feedback loop.

Abstract: This invention relates to the production and use of immortalized cell lines from primary chicken embryonic fibroblasts. The cells are useful as substrates for virus propagation, recombinant protein expression and recombinant virus production.