Abstract: A bacterial preparation capable of packaging phage .lambda. DNA is disclosed. This preparation is in lyophilized form and is stable at ambient temperature. In a preferred form, the preparation contains an over-expressed terminase protein, is prepared from the bacterial strain E. coli Cla [.lambda. c/857 Sam7 .DELTA.(cos-Nu1-A)::Kn.sup.r ]/.lambda. pTER and is capable of a packaging efficiency of at least 1.times.10.sup.8 pfu/.mu.g wild type .lambda. DNA. The present invention is also a method of creating a phage .lambda. DNA packaging extract comprising the steps of preparing a bacterial extract capable of packaging phage .lambda. and lyophilizing said extract.

Abstract: The present invention provides a recombinant infectious bovine rhinotracheitis virus which is characterized by an insertion of an expressible foreign DNA sequence encoding a polypeptide into the BamHI C fragment of the infectious bovine rhinotracheitis virus genome.

Abstract: In many separation techniques, such as field flow fractionation, liquid chromatography and electro-phoresis, chemical species form bands that migrate at different velocities. If the data-digitization rate and excitation intensity are both set to be optimal for the fastest migrating band, to compensate for different band velocities, both the data-digitization rate and the excitation intensity are decreased as a function of time by a factor equal to the migration time of the fastest migrating band to the separation time.

Abstract: The present invention provides a method for the rapid isolation and recovery of a desired target DNA or RNA molecules from a mixture or library containing such molecules. The method involves the use of biotinylated probes and enzymatic repair-cleavage to eliminate undesired library members from a sample.

Abstract: The present invention makes available methods and reagents for novel manipulation of nucleic acids. As described herein, the present invention makes use of the ability of intronic sequences, such as derived from group I, group II, or nuclear pre-mRNA introns, to mediate specific cleavage and ligation of discontinuous nucleic acid molecules. For example, novel genes and gene products can be generated by admixing nucleic acid constructs which comprise exon nucleic acid sequences flanked by intron sequences that can direct trans-splicing of the exon sequences to each other. The flanking intronic sequences can, by intermolecular complementation, form a reactive complex which promotes the transesterification reactions necessary to cause the ligation of discontinuous nucleic acid sequences to one another, and thereby generate a recombinant gene comprising the ligated exons.

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: A promoterless foreign gene is intergrated into the lac operon of Streptococcus thermophilus by transforming a host strain of Streptococcus thermophilus with a donor plasmid which does not replicate in the host strain. The donor plasmid includes a vector backbone and a sequence containing a foreign gene operably intergrated into at least a part of the lac operon of the host strain, in front of at least a part of the lacZ gene of the lac operon, and the sequence conserves the frame of the genomic lac operon of the host strain. Cointegrate transformants are identified in which the complete donor plasmid is integrated into the geonomic lac operon of the host strain, and an integrant transformant is then selected, the genome of which does not include the vector backbone of the donor plasmid but does include the foreign gene, which is operably integrated in front of the lacZ gene of the conserved genomic lac operon and which is stably maintained and expressed upon selective pressure on expression of the lacZ gene.

Abstract: Peptides that form lightly associated homodimers can be used to form dimers and multimers of other molecules and molecular motifs of interest. These association peptides can dimerize regardless of whether motifs are added to the amino-terminus of the peptide, or the carboxy terminus of the peptide, although additions to the carboxy-terminus of the association peptides require the presence of certain acidic residues.

Abstract: Novel compounds that mimic and/or modulate the activity of wild-type nucleic acids. In general, the compounds contain a selected nucleoside sequence wherein the nucleosides are covalently bound through linking groups that contain adjacent nitrogen atoms.

Abstract: The present invention provides vectors for the efficient and position-specific integration of expressible, exogenous nucleotide sequences into cellular genomes. This invention takes advantage of the discovery of a position-specific endonuclease and position-specific insertion markers for the design of said vectors. In addition, a gene comprising a recombinant nucleic acid molecule encoding a polypeptide possessing the biological activity of a position-specific endonuclease, wherein the biological activity of said endonuclease is the catalysis of position-specific insertion of genetic material carried between the position-specific integration markers, is disclosed.

Abstract: A bacteriophage packaging site-based vector system is describe that involves cloning vectors and methods for their use in preparing multiple copy number bacteriophage libraries of cloned DNA. The vector is based on a DNA segment that comprises a nucleotide sequence defining a bacteriophage packaging site located between two termini having ligation means used to concatamerize fragments of DNA having compatible ligation means. Methods for preparing a concatameric DNA for packaging cloned DNA segments, and for packaging the concatamers to produce a library are also described.

Abstract: Methods for the detection of nucleic acid using sandwich hybridization are provided. The target nucleic acids are captured by hybridization with oligonucleotides covalently attached to polystyrene solid supports to form complexes which are then hybridized to detection oligonucleotides to facilitate the analysis of the target nucleic acid sequences.

Abstract: Targeted gene insertion methodology can increase the production of an antimicrobial metabolite and comprises the steps of:a) identifying the gene in unaltered chromosomal DNA which encodes an enzyme that catalyzes the rate controlling step in a biosynthetic pathway in the production of increased concentration of a precursor to a core molecule which leads to the antimicrobial metabolite; andb) inserting into unaltered chromasomal DNA a genetic delivery vehicle (such as a vector, phage, or virus) which carries a gene which encodes the enzyme that catalyzes the rate controlling step in the biosynthesis in the production of a core molecule which is a precursor to the antimicrobial metabolite into unaltered chromosomal DNA, said delivery vehicle being compatible with said unaltered chromosomal DNA, the delivery vehicle being generated byinserting at least one exact or modified copy of the gene determining the rate controlling step into the compatible delivery vehicle, andinserting the delivery vehicle containing t

Abstract: Methods and compositions are described for making ribozymes which can release or activate molecules including autocatalytically replicatable RNA such as MDV-1.

Abstract: The present invention provides compositions and methods for inserting a copy of a heterologous gene into the chromosome of a host cell such as E. coli through the use of a chromosomal transfer DNA, a circular, non-self-replicating DNA carrying a site-specific recombination site such as lambda att P. The host cell chromosome contains a second site-specific recombination site. When the chromosomal transfer DNA is introduced into the host cell, expression of an integration enzyme such as integrase causes the integration of the chromosomal transfer DNA into the host cell chromosome at the second recombination site.

Abstract: A method is disclosed for automated multiplex sequencing of DNA with an integrated automated imaging hybridization chamber system. This system comprises an hybridization chamber device for mounting a membrane containing size-fractionated multiplex sequencing reaction products, apparatus for fluid delivery to the chamber device, imaging apparatus for light delivery to the membrane and image recording of fluorescence emanating from the membrane while in the chamber device, and programmable controller apparatus for controlling operation of the system. The multiplex reaction products are hybridized with a probe, then an enzyme (such as alkaline phosphatase) is bound to a binding moiety on the probe, and a fluorogenic substrate (such as a benzothiazole derivative) is introduced into the chamber device by the fluid delivery apparatus.

Abstract: A rapid method for transfection of a cell under physiological conditions suitable to the survival and growth of the cell is disclosed. According to the method, a stable nucleoprotein complex is provided. The nucleoprotein complex comprises a single-stranded DNA sequence in stable combination with RecA protein molecules. Cells to be transformed are cultured in a physiologically suitable medium to which the nucleoprotein complex has been added. As the cells grow and undergo mitosis, the nucleoprotein complex is taken up within some of the cells and becomes integrated into the genome. The method accomplishes transfection without resort to infectious vectors or permeabilization or other manipulation of the cell membrane. According to another object of the invention, a diagnostics method is provided. A directly detectable reporter label or an indirectly detectable ligand is bound to the nucleoprotein complex to provide a DNA probe which then is taken up into the cell and integrates into the cell's genome.

Abstract: A first polynucleotide molecule coding for a transactivator fusion protein comprising the tet repressor and a protein capable of activating transcription in eucaryotes. A second polynucleotide molecule coding for a protein, wherein said polynucleotide is operably linked to a minimal promoter operably linked to at least one tet operator sequence is also disclosed. A method to regulate the expression of a protein coded for by a polynucleotide, by cultivating the eucaryotic cell of the invention in a medium comprising tetracycline or a tetracycline analogue is also disclosed. Kits containing the polynucleotide molecules are also disclosed.

Abstract: Genes are disclosed which are capable of directing the synthesis in a selected host microorganism of two thaumatin I analogues both of which have the amino acid sequence of natural thaumatin I including an aspartate amino acid residue in the 113th position from the amino terminal end of the polypeptide and one of which additionally has a lysine amino acid residue substituted for asparagine in the 46th position from the amino terminal end.

Abstract: A method for making crystals of interferon alpha-2 and the use thereof in depot formulations are disclosed.