Abstract: Superfragments of chromosomal DNA having 1 million to 100 million base pairs may be formed by irradiating microcells containing a chromosome with a high dose of .gamma.-irradiation. Such superfragments are useful for the rapid cloning of genes.

Abstract: The present invention is directed to recombinant DNA vector molecules and uses therefor. More particularly, the present invention relates to the construction of recombinant vectors useful in the high frequency cloning of cDNA. The present invention also relates to vectors capable of directing the synthesis of single stranded cDNA and thereby permitting enrichment of specific sequences by hybridization/selection.

Abstract: The present invention provides improved methods for manipulating recombinant DNA in gene cloning and expression. More specifically, the invention provides methods capable of altering a nucleic acid sequence present at the termini of a target sequence.

Abstract: A screening method for the selection of mutagenized proteins that are normally secreted by cells is described. The method includes the development of a cloning vector for the expression of secretory proteins as fusion proteins on the cell surface of transfected mammalian cells. The secreted protein is displayed on the cell surface by fusion with the glycophospholipid membrane anchor of decay accelerating factor (DAF). Tissue-type plasminogen activator (t-PA), which is normally secreted, is used as a model protein. PCR mutagenesis is used to generate random mutations within the Kringle 1 (K1) domain of t-PA. Fluorescence activated cell sorting (FACS) is employed to screen for t-PA mutants possessing a loss of an epitope to a specific Mab, whose nonlinear binding domains overlap with the t-PA clearance receptor contact regions novel t-PA mutants designated N115S, N1425S, and K159R were discovered by this method.

Abstract: The present invention provides a recombinant fusion protein comprising an antigenic amino acid sequence fused to at least a portion of the gpX glycoprotein from pseudorabies virus. Such a protein may be formulated into a vaccine and delivered to an animal using a live herpesvirus vector adapted to express the fusion protein.

Abstract: The present invention contemplates a method of physiologic engineering by genetically altering second messenger levels in cells. This method allows the hyperactivation or inhibition of cell function within cells, tissues and animals by introducing a foreign gene that alters a second messenger system. The use of physiologically engineered animals as systems for determining the effectiveness of therapeutic compositions is also contemplated.

Abstract: A process using microorganisms which contain genes, which form an active xylene monooxygenase, which form no effective, chromosomally or plasmid-coded alcohol hydrogenase, and which are, thus, capable of hydroxylating methyl groups on aromatic 5- or 6-atom heterocycles to the corresponding hydroxymethyl derivatives, for the production of hydroxymethylated 5- or 6-atom heterocycles.

Abstract: The nucleotide sequence of the genome for bovine diarrhea virus (BDV) is disclosed. The sequence permits design and construction of vaccines effective against BDV.

Abstract: An rpoH gene that encodes a .sigma..sup.32 protein in which cysteine instead of arginine is at amino acid residue 268 was isolated from a mutant of Escherichia coli strain W3110. The rpoH gene has thymine instead of cytosine at the nucleotide position corresponding to 802 in the wild-type rpoH gene. Purified strains of E. coli W3110 and JM101 having a mutant rpoH gene in which thymine instead of cytosine is at the nucleotide position corresponding to 802 in a wild-type rpoH gene and a method for synthesis of proteins at enhanced levels are also disclosed. The method comprises introducing an expression vehicle into an E. coli strain containing the mutant rpoH gene of this invention.

Abstract: The invention relates to an expression vector system based on the regulation of bacterial luminescence (the lux gene system). The invention further relates to the construction of a precisely regulatable expression vector system which comprises a complete luxR gene in combination with an inactivated luxI gene. If the system is turned off, no significant transcription occurs of any cloned gene product when used in combination with the regulatory scheme of the invention as is demonstrated by using the bacteriophage .lambda. lysis genes. The induction of transcription relies on the addition of exogenous autoinducer which is both inexpensive and easy-to-use and which is required in only minute amounts.

Abstract: Methods of designing or modifying protein structure at the protein or genetic level to produce specified amino-termini in vivo or in vitro are described. The methods can be used to alter the metabolic stability and other properties of the protein or, alternatively, to artificially generate authentic amino-termini in proteins produced through artificial means. The methods are based upon the introduction of the use of artificial ubiquitin-protein fusions, and the discovery that the in vivo half-life of a protein is a function of the amino-terminal amino acid of the protein.

Abstract: Viable bacteria may be detected in biological samples by exposing bacterial cultures obtained from the samples to transducing particles having a known host range. Such transducing particles carry a heterologous gene capable of altering the phenotype of the bacteria in a readily detectable manner. For example, the transducing particles may carry an ice nucleation gene and the alteration of phenotype may be detected using an ice nucleation assay. By employing a panel of phage, unknown bacteria nmay be typed based on the pattern of reactivity observed. The transducing particles may be prepared by introducing a synthetic transposable element carrying the heterologous gene to a host carrying a prophage having the desired host range. After transposition, the host may be induced to a lytic cycle to release the transducing particles carrying the heterologous gene.

Abstract: A genetic element comprising an expression vector and a gene coding for transketolase is utilized to enhance diversion of carbon resources into the common aromatic pathway.

Abstract: The present invention provides improved methods for manipulating recombinant DNA in gene cloning and expression. More specifically, the invention provides methods capable of altering a nucleic acid sequence present at the termini of a target sequence.