Abstract: DNA is efficiently transformed into a host by electroporation in the presence of a methylation package, which greatly improves the efficiency of the transformation. The methylation package comprises a source of cysteine, such as cysteine, homocysteine, or glutathione, with optional iron and magnesium ions.

Abstract: The invention relates to a method for influencing the p53 binding to a target gene, wherein the conformations of p53 and the target gene are coordinated especially by means of conformation modulators and the binding of p53 can be directly or indirectly detected.

Abstract: A novel co-culture system using human brain endothelial cells (HUBEC) which promotes the expansion of human CD34+CD38− cells consistent with the PMVEC system is disclosed. HUBEC were isolated from cadaveric donors, passed in primary culture, cloned and found to be Von Willebrand Factor positive. Cultivation of purified bone marrow CD34+ cells on HUBEC monolayers supplemented with GM-CSF+IL-3+IL-6+SCF+flt-3 ligand caused a 14.5-fold increase in total cells, an 6.6-fold increase in CD34+ cells, and, most remarkably, a 440-fold increase in CD34+CD38− cells after 7 days. Further, CFU-GM production increased 15.1-fold, BFU-E increased 8-fold, and CFU-Mix increased 5.2-fold. Optimal generation was dependent upon the continued presence of exogenous supplied cytokines. Moreover, we found that non-brain human endothelial cells isolated from the same donors supported neither the expansion nor the maintenance of human CD34+CD38− cells.

Abstract: The present invention provides a method of isolation of compounds that bind specifically to RNA or that alter the affinity of known compounds for RNA. The method utilizes changes in the levels of a fluorescent marker protein expressed in the same cell as a putative RNA-binding compound to detect those compounds of interest.

Abstract: The present invention provides a DNA molecule comprising a radiation responsive enhancer-promoter operatively linked to an encoding region that encodes at least one polypeptide. An encoding region can comprise a single encoding sequence for a polypeptide or two or more encoding sequences encoding DNA binding, activation or repression domains of a transcription factor. Processes for regulating polypeptide expression and inhibiting tumor growth using such DNA molecules are also provided.

Abstract: Disclosed is a process for identifying clones having a specified activity of interest, which process comprises (i) generating one or more expression libraries derived from nucleic acid directly isolated from the environment; and (ii) screening said libraries utilizing a fluorescence activated cell sorter to identify said clones. More particularly, this is a process for identifying clones having a specified activity of interest by (i) generating one or more expression libraries derived from nucleic acid directly or indirectly isolated from the environment; (ii) exposing said libraries to a particular substrate or substrates of interest; and (iii) screening said exposed libraries utilizing a fluorescence activated cell sorter to identify clones which react with the substrate or substrates.

Abstract: The subject invention lies in the field of microorganism mutation and selection of the mutants. In particular, the invention is directed at obtaining metabolic mutants in a simple, direct and specific manner. In a preferred embodiment it is also possible to obtain desired mutants not comprising recombinant DNA, thereby facilitating incorporation thereof in products for human consumption or application, due to shorter legislative procedures. The method according to the invention involves random mutation and specific selection of the desired metabolic mutant. Knockout mutants wherein a gene associated with metabolism is absent or inactivated and mutants with increased or decreased DNA binding capacity are also claimed.

Abstract: The present invention provides a method of efficiently screening for steroid sulfatase activity-inhibiting compounds which are useful for the treatment of hormone-dependent diseases such as breast cancer, etc. by causing a test compound to act on cells carrying an introduced steroid sulfatase gene and estimating the inhibiting activity of the compound on the growth of the cells.

Abstract: An improved method of gene targeting, referred to as PCR-based gene targeting is disclosed, which generates cell lines or mice in which at least one allele of a specific gene is disrupted by double homologous recombination of a PCR-derived targeting vector with chromosomal DNA. The method is especially applied to murine macrophage cytokine-inducible nitric oxide synthase (MøiNOS).

Abstract: A method is provided for forming a graft in heart tissue which comprises the transplantation of cells chosen from cardiomyocytes, fibroblasts, smooth muscle cells, endothelial cells and skeletal myoblasts. The grafts are especially useful in treating scar tissue on the heart.

Abstract: Yeast strains expressing human genes coding regulating cytochrome P450 expression, a method for making same and applications thereof are described.

Abstract: Nucleic acid constructs containing hybrid promoters for use in gene therapy and genetic manipulation. The invention relates to a nucleic acid construct for the precise, regulated expression of genes in host cells, which construct exhibits at least one mutation which inhibits the proper expression of the expressed gene and exhibits at least one additional second mutation which relieves the inhibition due to the first mutation, to an isolated cell which harbors the nucleic acid construct, and to the use of the nucleic acid construct for preparing pharmaceuticals and for treating diseases with excessive cell proliferation.

Abstract: Dominant-negative alleles of human mismatch repair genes can be used to generate hypermutable cells and organisms. By introducing these genes into mammalian cells new cell lines with novel and useful properties can be prepared more efficiently than by relying on the natural rate of mutation or introduction of mutations by chemical mutagens. These methods are useful for generating novel and highly active antimicrobial molecules as well as superior antimicrobial agents from pre-existing chemicals. These methods are also useful for generating cell lines expressing novel antimicrobials that are useful for pharmaceutical manufacturing.

Abstract: Methods for the rapid repression of gene function in eucaryotic cells are disclosed including inducible means for both shutting down a targeted gene's transcription and rapidly removing a targeted gene's polypeptide product.

Abstract: The present invention focuses on the super-expression of foreign genes in transgenic cells by combining within a single cDNA construct and respective RNA transcript, several trans- and cis-acting genetic elements of viral origin which act in concert.

Abstract: The present invention relates to the development of a positive selection vector based on insertional reconstruction of a reporter gene or of a regulatory gene controlling the expression of a reporter gene. The cloning vector carries a reporter gene or a regulatory gene with a mutation rendering the reporter or the regulatory gene protein functionally inactive. A primer carrying a nucleic acid sequence that corrects the mutation is used during PCR amplification of a targeted nucleic acid sequence, and the amplified DNA fragment is then ligated to the said vector thus reconstructing the wild-type reporter or regulatory gene.

Abstract: A solution containing double stranded nucleic acid is heated, thereby dissociating the strands, which in turn are cooled and rebounded. A homoduplex and a heteroduplex are formed when the double stranded nucleic acid is a hetero body, while only a homoduplex is formed when the double stranded nucleic acid is a homo body. The temperature of the solution is increased while measuring ultraviolet absorption, thereby acquiring a thermal melting profile. When the heated and cooled solution contains only a homoduplex, the thermal melting profile has a single Tm temperature or two Tm temperatures close to each other. When the heated and cooled solution contains a homoduplex and a heteroduplex, the thermal melting profile has two separate Tm temperatures or further single Tm temperatures close to each other. The method therefore allows for the detection of a heteroduplex or a mutation in a sample double stranded nucleic acid.

Abstract: The JeT promoter is a recombinant promoter with transcriptional activity comparable to a number of strong mammalian promoters. The promoter consists of five key elements: (1) a TATA box; (2) a transcription initiation site (Inr); (3) a CAT consensus sequence in conjunction with (4) a CArG element and finally, (5) four Sp1 transcription binding sites (GGGCGG) arranged in two tandems.

Abstract: The present invention provides the promoter clone discovery of an alpha-amylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated alpha-amylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.

Abstract: The present invention is directed to nucleic acid sequences encoding a novel regulatory element that induces a high level of expression to operably linked genes, upon exposure to hypoxic conditions. Furthermore, the regulatory element can be used to prepare an expression vector for transforming plant cells, wherein the DNA construct comprises a hypoxia inducible promoter operably linked to a gene sequence.