Abstract: The present invention provides an isolated, purified and characterized human tyrosine hydroxylase (hTH) promoter nucleic acid sequence. The invention further provides a method of selecting TH positive (TH+) cells by preparing a construct comprising a hTH promoter operably linked to a heterologous nucleic acid sequence, for example, green fluorescent protein encoding sequence, and transfecting cells, particularly stem cells, with the construct. The invention also provides a hTH promoter, useful in gene therapeutic applications in driving therapeutic genes or other nucleic acid sequences operably linked to the hTH promoter. Additionally, the invention provides cell lines and transgenic animals expressing a transgene comprising the hTH promoter operably linked to a heterologous sequence, which cell lines and transgenic animals are useful for isolating TH+ cells for transplantation or for screening of therapeutic agents that affect TH+ function.

Abstract: A method for introducing and expressing genes in animal cells, in which the animal cells are transfected with bacterial blebs containing a eukaryotic expression cassette encoding the gene. Bacterial blebs comprising a eukaryotic expression cassette, wherein the bacterial blebs are derived from gram negative bacteria.

Abstract: The present invention is directed to methods and compositions for use of homologous recombination for directed evolution, gene reassembly, and directed mutagenesis. One aspect of the present invention relates to methods for use of bacterial conjugative transfer and homologous recombination for directed evolution, gene reassembly, and directed mutagenesis. Another aspect of the present invention relates to compositions for use in or produced by the methods of the present invention, including libraries, archives and databases.

Abstract: A method is described for the rapid identification and isolation of cells based on the presence or absence of an ectopically-expressed N-acetyllactosaminide 3-? Galactosyltransferase (?GT) enzyme for the production of ?Galactosyl-(1,3)Galactosyl (?Gal) epitopes on the surface of ?Gal-negative cells. These cells which are genetically modified to express the ?GT enzyme and ?Gal epitopes on glycosylated lipids and proteins of the cell surface are then labeled via an antibody composition which recognizes and binds the ?Gal epitopes on the cell surface. Cells labeled with the anti-?Gal antibody can be isolated by sorting via fluorescence activated cell sorting (FACS), or by magnetic panning techniques. This method is suitable for the rapid positive or negative selection of ?Gal-positive cells from within a population of ?Gal-negative cells without the need to expose cells to antibiotics for any period of time.

Abstract: Disclosed is the isolation and characterization of EI24, a gene whose 2.4 kb mRNA is induced following etoposide treatment. Induction of EI24 mRNA by etoposide required expression of wild-type p53. Overexpression of functional p53 was sufficient to induce expression of the EI24 mRNA. The EI24 mRNA was also induced in a p53-dependent manner by ionizing irradiation of primary murine thymocytes. The invention is thus directed to an isolated EI24 protein, nucleotide sequences coding for and regulating expression of the protein, antibodies directed against the protein, and recombinant vectors and host cells containing the genetic sequences coding for and regulating the expression of the protein sequence. Antibodies can be used to detect EI24 in biological specimens, including, for example, human tissue samples.

Abstract: The invention relates to a method for the stable inversion of a DNA fragment upon recombinase-mediated rearrangements using two sets of two incompatible site-specific recombinase targeting sites (SSRTS) in the same order but in reverse orientation flanking the DNA fragment to be inverted. The invention also relates to a method for the stable inversion of the DNA fragment upon rearrangement mediated by a recombinase such as Cre recombinase. The invention also relates to a method for obtaining a transgenic cell of which at least one allele of a DNA sequence of interest is invalidated by a process of conditional deletion and the genome of which has a reporter gene inserted at the place of the DNA fragment deleted by the process of conditional deletion. A method to generate targeting sites to perform site-specific recombination mediated cassette exchange is also provided.

Abstract: The invention discloses proteins having ? (1–3)glucanosyltransferase type activities. These proteins can be used for detecting the antifungal activity of molecules.

Abstract: An antibiotic-independent vector for stable vector maintenance and high protein expression and a method for gene expression using this vector are disclosed. The stable maintenance of the vector is due to expression of a glutamic acid racemase to complement a D-glutamic acid auxotroph. No antibiotics, such as ampicillin, are necessary for the stable maintenance of this vector.

Abstract: Disclosed are methods of preventing or treating cardiac arrhythmia. In one embodiment, the methods include administering to an amount of at least one polynucleotide that modulates an electrical property of the heart. The methods have a wide variety of important uses including treating cardiac arrhythmia.

Abstract: Vectors for producing polypeptides heterologous to prokaryotes are described comprising, along with the polypeptide-encoding nucleic acid, anti-termination nucleic acid that inhibits intragenic transcription termination with a non-lambda promoter therefor and/or nucleic acid encoding a GreA or GreB protein and a promoter therefor. Also described are processes for producing a heterologous polypeptide in prokaryotic host cells utilizing such elements to improve the quality and/or quantity of heterologous polypeptide produced.

Abstract: The invention discloses proteins having ?(1-3)glucanosyltransferase type activities. These proteins can be used for detecting the antifungal activity of molecules.

Abstract: A method for producing recombinant adeno-associated virus in the absence of contaminating helper virus or wild-type virus involves culturing a mammalian host cell containing an rAd/AAV hybrid virus, an AAV rep sequence and an AAV cap sequence under the control of regulatory sequences directing expression thereof. The rAd/AAV hybrid virus contains a rAAV construct to be packaged into an AAV virion in an backbone containing the adenoviral sequences necessary to express E1a and E1b gene products and to permit replication of the hybrid virus. The method of the invention permits replication of the hybrid virus and production of rAAV virion in this host cell in the absence of a helper virus and obviates a subsequent purification step to purify rAAV from contaminating virus.

Abstract: Methods for delivering a biologically active molecule into a cell by linking a molecule to the cell surface, wherein the molecule can act as a surface receptor, then complexing the biologically active molecule with a ligand for the surface receptor, and finally contacting the biologically active molecule-ligand complex with the cell surface are disclosed. Delivery of any biologically active molecule, e.g. proteins, enzymes, nucleic acids, hormones, nucleic acids, and oligonucleotides, is contemplated. The use of biotin or biotinylated antibodies as the surface receptor is disclosed. The use of PEI and PEI-avidin conjugates complexed with oligonucleotides for delivery into a directly or indirectly biotinylated cell surface, along with the PEI-avidin-nucleic acid compositions, are disclosed. Primary and cultured cells with a covalently linked surface receptor molecule, such as biotin, on their surfaces are also disclosed.