Abstract: This invention pertains to BIV constructs encompassing BIV combination vectors, BIV vectors and BIV packaging vectors and particularly the invention pertains to a three vector system comprising: a) a BIV vector construct including a DNA segment from a BIV genome, a packaging sequence to package RNA into virions; a promoter operably linked to the DNA segment; and a transgene operably linked to a second promoter; b) a BIV packaging vector construct comprising a BIV DNA sequence fragment comprising at least a gag gene or pol gene of BIV; a promoter operably linked to the BIV DNA fragment; and a polyadenylation sequence located downstream of the BIV DNA fragment; and c) an expression vector construct comprising a gene encoding a viral surface protein. Also provided is a method for transferring a gene of interest into a mammalian cell.

Abstract: The present invention provides a general approach for G protein coupled receptors that may be used to define agonists and antagonists, and the specificity of receptor coupling to G protein subunits. Methods of the present invention use small volumes (microliters) and are compatible with high throughput flow cytometry. When assays of the present invention are multiplexed, the specificity of the interactions of a receptor with many G proteins may be determined simultaneously.

Abstract: Nucleic acid labeling compounds containing heterocyclic derivatives are disclosed. The heterocyclic derivative containing compounds are synthesized by condensing a heterocyclic derivative with a cyclic group (e.g. a ribofuranose derivative). The labeling compounds are suitable for enzymatic attachment to a nucleic acid, either terminally or internally, to provide a mechanism of nucleic acid detection.

Abstract: This invention pertains to BIV constructs encompassing BIV combination vectors, BIV vectors and BIV packaging vectors and particularly the invention pertains to a three vector system comprising: a) a BIV vector construct including a DNA segment from a BIV genome, a packaging sequence to package RNA into virions; a promoter operably linked to the DNA segment; and a transgene operably linked to a second promoter; b) a BIV packaging vector construct comprising a BIV DNA sequence fragment comprising at least a gag gene or pol gene of BIV; a promoter operably linked to the BIV DNA fragment; and a polyadenylation sequence located downstream of the BIV DNA fragment; and c) an expression vector construct comprising a gene encoding a viral surface protein. Also provided is a method for transferring a gene of interest into a mammalian cell.

Abstract: A nucleic acid sequence required for regulating the autolytic activity of bacteria is provided. Also provided are polypeptides encoded by the gene or mutant gene as well as vector and host cells for expressing these polypeptides. Methods for identifying and using agents which interact with the gene or mutant gene or polypeptides encoded thereby to inhibit bacterial growth and infectivity are also provided.

Abstract: The present invention relates to multivalent Fvantibody construct having at least four variable domains which are linked with each over via the peptide linkers 1, 2 and 3. The invention also concerns expression plasmids which code for such an Fvantibody construct and a method of producing the Fvantibody constructs as well as their use.

Abstract: The present invention is directed to a transformation systems and vectors for making transgenic organisms that includes a vector containing a modified piggyBac transposon into which is inserted at least one fluorescent protein gene linked to a polyubiquitin promoter sequence. A helper transposase vector that includes an hsp70 promoter sequence upstream of the putative piggyBac promoter that increases the transformation frequency of this system can also be included.

Abstract: This invention pertains to BIV constructs encompassing BIV combination vectors, BIV vectors and BIV packaging vectors and particularly the invention pertains to a three vector system comprising: a) a BIV vector construct including a DNA segment from a BIV genome, a packaging sequence to package RNA into virions; a promoter operably linked to the DNA segment; and a transgene operably linked to a second promoter; b) a BIV packaging vector construct comprising a BIV DNA sequence fragment comprising at least a gag gene or pol gene of BIV; a promoter operably linked to the BIV DNA fragment; and a polyadenylation sequence located downstream of the BIV DNA fragment; and c) an expression vector construct comprising a gene encoding a viral surface protein. Also provided is a method for transferring a gene of interest into a mammalian cell.

Abstract: A display and method of preparing 7-transmembrane and other receptors for real-time kinetic analysis of binding interactions. The invention includes display on beads and in micelles for multi-well and flow cytometric analysis. The invention is useful for ligand discovery and drug action discovery, and G-protein response in particular.

Abstract: The invention concerns the use of stabilized oligonucleotides comprising at least an octamer motif of the type: 5?-purine-purine-CG-pyrimidine-pyrimidine-X1X2-3? wherein the pair X1-X2- is AT, AA, CT or TT, for preparing a medicine with antitumor activity.

Abstract: LbpB polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing LbpB polypeptides and polynucleotides in the design of protocols for the treatment of neisserial disease, among others, and diagnostic assays for such conditions.

Abstract: A retroviral vector for gene reconstitution is provided that includes a 3? portion of a heterologous nucleic acid sequence 5? of a first att site and a 5? portion of the heterologous nucleic acid sequence 3? of a second att site. A sub-portion of the 3? portion of the heterologous nucleic acid sequence and a sub-portion the 5? portion of the heterologous nucleic acid sequence are direct repeats. A retroviral vector for gene reconstitution is also provided that includes a 3? portion of a heterologous nucleic acid sequence inserted into or adjacent to a 5? retroviral terminal repeat of the retroviral vector, and a 5? portion of the heterologous nucleic acid sequence inserted into or adjacent to a 3? retroviral terminal repeat of the retroviral vector, wherein the 3? and the 5? retroviral terminal repeats each comprise an att site. Methods and kits are also provided.

Abstract: Proteinase inhibitors comprising a Kunitz domain are disclosed. The Kunitz domain comprises a sequence of amino acid residues as shown in SEQ ID NO:3, wherein the sequence is at least 80% identical to residues 6 through 56 of SEQ ID NO:2. Also disclosed are methods for making the proteinase inhibitors, and expression vectors and cultured cells that are useful within the methods. The proteinase inhibitors may be used as components of cell culture media, in protein purification, and in certain therapeutic and diagnostic applications.

Abstract: The present invention relates to an expression system for the expression of proteins and peptides in a methanotrophic bacterium, preferable the M. capsulatus. Further, the invention relates to the exportation and display of said peptides and proteins on the surface of said bacteria. The invention also describes a method for the production of a desired protein in the M. capsulatus.

Abstract: The invention provides BASB033 polypeptides and polynucleotides encoding BASB033 polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are diagnostic, prophylactic and therapeutic uses.

Abstract: The present invention relates to means for cleaving a nucleic acid cleavage structure in a site-specific manner. Enzymes, including 5? nucleases and 3? exonucleases, are used to detect and identify nucleic acids derived from microorganisms. Methods are provided which allow for the detection and identification of bacterial and viral pathogens in a sample.

Abstract: To provide DNA comprising mutant FRT sequence which causes recombination reaction between two mutant FRT sequences having an identical sequence to each other but does not cause recombination reaction with a wild-type FRT sequence, in the presence of FLP recombinase; and a method for performing high-efficiency, gene insertion or gene replacement. A DNA comprising a mutant FRT sequence. A DNA comprising a mutant FRT sequence possessing (A) causing no specific DNA recombination reaction with wild type FRT, even if FLP recombinase is present, and (B) causing specific DNA recombination reaction with another mutant FRT sequence having an identical sequence thereto in the presence of recombinase FLP; gene replacement method using the DNA in the presence of recombinase FLP; and a specific DNA recombination method, characterized in that a specific DNA recombination reaction is carried out by using two mutant FRT sequences in the presence of recombinase FLP.

Abstract: The present invention provides novel isolated nucleic acids encoding antigenic proteins derived from Sarcocystis neurona, or unique fragments thereof. In particular, the invention provides novel isolated nucleic acids encoding membrane-associated polypeptides SnSAG2, SnSAG3, and SnSAG4. Also provided are purified antigenic polypeptide fragments encoded by the novel nucleic acid sequences set forth herein that encode for SnSAG2, SnSAG3, and SnSAG4. Also provided are isolated nucleic acids capable of selectively hybridizing with the nucleic acid from Sarcocystis neurona. The invention also provides vectors comprising the nucleic acids of the invention encoding an antigenic protein derived from Sarcocystis neurona or a unique fragment thereof and provides the vector in a host capable of expressing the polypeptide encoded by that nucleic acid.

Abstract: Current technologies allow the generation of artificial DNA molecules and/or the ability to alter the DNA sequences of existing DNA molecules. With a careful coding scheme and arrangement, it is possible to encode important information as an artificial DNA strand and store it in a living host safely and permanently. This inventive technology can be used to identify origins and protect R&D investments. It can also be used in environmental research to track generations of organisms and observe the ecological impact of pollutants. Today, there are microorganisms that can survive under extreme conditions. As well, it is advantageous to consider multicellular organisms as hosts for stored information. These living organisms can provide as memory housing and protection for stored data or information. The present invention provides well for data storage in a living organism wherein at least one DNA sequence is encoded to represent data and incorporated into a living organism.

Abstract: This invention provides an isolated mammalian nucleic acid molecule encoding an alternatively spliced prostate-specific membrane (PSM?) antigen. This invention provides an isolated nucleic acid molecule encoding a prostate-specific membrane antigen promoter. This invention provides a method of detecting hematogenous micrometastic tumor cells of a subject, and determining prostate cancer progression in a subject.

Abstract: Method and apparatus for constructing a cDNA library by hybridizing mRNA with oligo (dT) on a support and treating with a reverse transcriptase to immobilized complementary DNA, or for constructing a gDNA library by ligating a double-stranded chromosomal DNA library with an oligonucleotide on a support having a restriction enzyme site and then immobilizing the gDNA library.

Abstract: The invention relates to a process for the controlled amplification of at least one part of a starting DNA containing a plurality of restriction sites for a determined specific restriction endonuclease, and of which at least part or its nucleic acid is unknown. Application of this process to human, animal or plant DNA fingerprinting, to identification of restriction fragment length polymorphisms. Kit for the application of the process.

Abstract: A display and method of preparing 7-transmembrane and other receptors for real-time kinetic analysis of binding interactions. The invention includes display on beads and in micelles for multi-well and flow cytometric analysis. The invention is useful for ligand discovery and drug action discovery, and G-protein response in particular.

Abstract: Methods are provided for producing a vector that includes at least one splicable intron. In the subject methods, intron containing vectors are produced from donor and acceptor vectors that each include a site specific recombinase site, where the subject donor and acceptor vectors further include splice donor and acceptor sites that, upon site specific recombination of the donor and acceptor vectors, define an intron in the product vector of the recombination step. Also provided are compositions for use in practicing the subject methods, including the donor and acceptor vectors themselves, as well as systems and kits that include the same. The subject invention finds use in a variety of different applications, including the production of expression vectors that encode C-terminal tagged fusion proteins, the production of expression vectors that encode pure protein and not a fusion thereof, and the like.

Abstract: The present invention relates generally to compositions and methods for enhancing recombinational cloning of nucleic acid molecules. In particular, the invention relates to compositions comprising one or more ribosomal proteins and one or more additional protein components required for recombinational cloning. More particularly, the invention relates to such compositions wherein the ribosomal proteins are one or more E. coli ribosomal proteins, still more particularly wherein the ribosomal proteins are selected from the group of E. coli ribosomal proteins consisting of S10, S14, S15, S16, S17, S18, S19, S20, S21, L20, L21, and L23 through L34, and most particularly S20, L27, and S15. The invention also relates to the use of these compositions in methods for recombinational cloning of nucleic acids, in vitro and in vivo, to provide chimeric DNA molecules that have particular characteristics and/or DNA segments.

Abstract: A unique HCV RNA molecule is provided having an enhanced efficiency of establishing cell culture replication. Novel adaptive mutations have been identified within the HCV non-structural region that improves the efficiency of establishing persistently replicating HCV RNA in cell culture. This self-replicating polynucleotide molecule contains, contrary to all previous reports, a 5?-NTR that can be either an A as an alternative to the G already disclosed and therefore provides an alternative to existing systems comprising a self-replicating HCV RNA molecule. The G->A mutation gives rise to HCV RNA molecules that, in conjunction with mutations in the HCV non-structural region, such as the G(2042)C/R mutations, possess greater efficiency of transduction and/or replication. These RNA molecules when transfected in a cell line are useful for evaluating potential inhibitors of HCV replication.

Abstract: The present invention relates to a new tumor suppressor, designated CAR-1, the gene for which is located on the short arm of human chromosome 1. This gene is directly implicated in colon, kidney and breast cancers, and the CAR-1 ubiquitous expression of the corresponding transcript suggests that it may be involved in yet others. Thus, one aspect of the invention is the diagnosis of CAR-1-related malignancies. The full length cDNA for CAR-1, as well as oligonucleotides derived therefrom, are disclosed. Screening methods for modulators of CAR-1 function and expression, as well as methods for cancer therapy, are described.

Abstract: A helper dependent adenoviral vector system is provided. The subject helper dependent adenoviral vector system is made up of: (1) a “gutless” adenoviral vector that include cis-acting human stuffer DNA that provides for in vivo long term, high level expression of a coding sequence present on the vector; (2) an adenoviral helper vector that is characterized by having an adenoviral genome region flanked by recombinase recognition sites, where the helper vectors further include a non-mammalian endonuclease recognition site positioned outside of the adenoviral genome region; and (3) a mammalian cell that expresses the corresponding recombinase and endonuclease, as well as the adenoviral preterminal and polymerase proteins. Also provided are methods of using the subject systems to produce virions having the subject helper dependent adenoviral vectors encapsulated in an adenoviral capsid. In addition, kits for use in practicing the subject methods are provided.

Abstract: Presented are methods and compositions for targeted chromosomal genomic alterations using modified single-stranded oligonucleotides. The oligonucleotides of the invention have at least one modified nuclease-resistant terminal region comprising phosphorothioate linkages, LNA analogs or 2?-O-Me base analogs.

Abstract: The present invention provides nucleic acid molecules which may be used as standards for estimating the size (in base pairs) and mass of linear, double-stranded or single-stranded nucleic acid molecules separated by size. The nucleic acid molecules of the invention may be DNA molecules, RNA molecules or DNA/RNA hybrid molecules, and may be double-stranded or single-stranded. The invention also provides methods for producing nucleic acid sizing ladders from these nucleic acid molecules, ladders produced by such methods, and methods for estimating the size and mass of nucleic acid molecules by comparison to these nucleic acid sizing ladders.

Abstract: A highly homogeneous library can be obtained by cleaving a genomic DNA by a sequence-independent cleavage method, such as sonication. By selecting satellite sequences from such a library, efficiency of isolation is improved. Thus, an efficient method of isolating microsatellite sequences, which are useful as DNA markers, is provided.

Abstract: The invention provides polyamide-oligonucleotide derivatives of the formula: F[(DNA-Li)q(PNA-Li)r(DNA-Li)s(PNA)t]xF?. In the formula, q, r, s, and t are, independently of one another, zero or 1, where the total of two or more adjacent q, r, s, and t is greater than or equal to 2; and x is 1 to 20. In the formula, DNA is a nucleic acid such as DNA or RNA or a known derivative thereof. Li is a covalent linkage between DNA and PNA, where the covalent linkage comprises a bond or an organic radical with at least one atom from the series consisting of C, N, O, or S. PNA is a polyamide structure which contains at least one nucleotide base that is different from thymine. F and F? are end groups and/or are linked together by a covalent bond. The invention also provides physiologically tolerated salts of the above formula. The invention further provides a process for preparation of the polyamide-oligonucleotide derivatives of the invention as well as their use as pharmaceuticals, as gene probes, and as primers.

Abstract: The present invention provides compositions, methods, and kits for covalently linking nucleic acid molecules. The methods include a strand invasion step, and the compositions and kits are useful for performing such methods. For example, a method of covalently linking double stranded (ds) nucleic acid molecules can include contacting a first ds nucleic acid molecule, which has a topoisomerase linked to a 3? terminus of one end and has a single stranded 5? overhang at the same end, with a second ds nucleic acid molecule having a blunt end, such that the 5? overhang can hybridize to a complementary sequence of the blunt end of the second nucleic acid molecule, and the topoisomerase can covalently link the ds nucleic acid molecules. The methods are simpler and more efficient than previous methods for covalently linking nucleic acid sequences, and the compositions and kits facilitate practicing the methods, including methods of directionally linking two or more ds nucleic acid molecules.

Abstract: Cytomegalovirus (CMV) Intron A fragments for expressing gene products are disclosed. Also described are expression vectors including the fragments, as well as methods of using the same.

Abstract: Cloning and expression of genes encoding H. somnus transferrin-binding proteins are described. The transferrin-binding proteins can be used in vaccine compositions for the prevention and treatment of H. somnus infections, as well as in diagnostic methods for determining the presence of H. somnus infections.

Abstract: Methods of cloning and/or amplifying toxic genes in bacteria using a vector which amplifies the toxic gene in bacteria and also allows subsequent expression in mammalian systems is provided. A vector having an origin of replication, a first promoter, a polylinker, a second promoter in reverse orientation with respect to the first promoter, a poly adenylation signal, and a gene encoding a selectable marker, and optionally an enhancer operably connected to the first promoter, and/or a nucleotide sequence encoding a toxic protein is also provided.

Abstract: The present invention provides methods for determining the genotype of a nucleic acid at the site of a polymorphism. The methods achieve sensitivities great enough to detect the presence of any difference between the nucleic acids, even single nucleotide polymorphisms. In the methods, the nucleic acid is compared to a reference nucleic acid having a known genotype. The nucleic acids can be of any length, even less than 100 base pairs. In methods, one or more extra mismatches are introduced into the nucleic acids at or near the site of the polymorphism. The nucleic acids are contacted under conditions in which they are capable of forming a stable four-way complex that can be detected to indicate that the nucleic acids differ in genotype.

Abstract: The invention is directed to polynucleotide constructs having at least one transcriptional enhancer operably liked to a rice EPSPS promoter and to a sequence encoding the rice EPSPS chloroplast transit peptide and rice EPSPS enzyme, the encoded enzyme having a modified region that confers herbicide resistance.

Abstract: This invention pertains to BIV constructs encompassing BIV combination vectors, BIV vectors and BIV packaging vectors and particularly the invention pertains to a three vector system comprising: a) a BIV vector construct including a DNA segment from a BIV genome, a packaging sequence to package RNA into virions; a promoter operably linked to the DNA segment; and a transgene operably linked to a second promoter; b) a BIV packaging vector construct comprising a BIV DNA sequence fragment comprising at least a gag gene or pol gene of BIV; a promoter operably linked to the BIV DNA fragment; and a polyadenylation sequence located downstream of the BIV DNA fragment; and c) an expression vector construct comprising a gene encoding a viral surface protein. Also provided is a method for transferring a gene of interest into a mammalian cell.

Abstract: The present invention relates to a process for amplifying DNA of an organism. More particularly, the present invention is directed to a process for amplifying DNA of an organism through Polymerase Chain Reaction(PCR) without any information regarding a primer needed for amplifying DNA of an organism.

Abstract: Protective high molecular weight (HMW) proteins are produced recombinantly by expression from E. coli by using a promoter effective in E. coli and a nucleic acid molecule which contains a modified operon of a non-typeable strain of Haemophilus. The modified operon contains the portion only of the A region which encodes the mature HMW protein and the complete B and C regions of the operon. Enhanced levels of expression of the HMW proteins can be achieved by including the E. coli cer gene, a further copy of the portion of the A region of the operon encoding the mature protein or both in the expression vector. Nucleotide and deduced amino acid sequences of the hmw1 and hmw2 genes and HMW1 and HMW2 proteins, respectively of several non-typeable Haemophilus influenzae strain have been identified.

Abstract: Nucleic acid labeling compounds containing heterocyclic derivatives are disclosed. The heterocyclic derivative containing compounds are synthesized by condensing a heterocyclic derivative with a cyclic group (e.g. a ribofuranose derivative). The labeling compounds are suitable for enzymatic attachment to a nucleic acid, either terminally or internally, to provide a mechanism of nucleic acid detection.

Abstract: The invention is directed to methods for purifying Troponin I, particularly recombinant Tropnin I produced in a bacterial expression system. Recombinant Tropnin I can be advantageously purified after reversibly protecting the free sulfhydryl groups, e.g., by forming sulfates. In a specific example, Tropnin I reacted with sodium tetrafhionate yielded sulfitolyzed Tropnin I, which was purified by chromatography on an anion exchanger, followed by hydrophobic interaction chromatography. Facile deprotection of the sulfhydryl groups yields a highly purified product ready for refolding.

Abstract: This invention provides a method for identifying one or more complexes from a library of complexes, wherein said complex or complexes are selected for their ability to perform a preselected or desired function on a target molecule or by having a pre-selected structure, each complex being designated a morphatide, said method comprising: (a) preparing a library of morphatides, comprised of: (i) a scaffolding component selected from the group consisting of nucleic acid, nucleic acid like molecule or nucleic acid analog having one or more regions of randomized sequence; (ii) one or more linker components; and (iii) one or more agent molecules or type of agent molecules, linked to the scaffolding component by one or more type of linker components; and (b)screening the library of morphatides prepared in step (a) by contacting, binding, or associating the morphatides with one or more suitable target molecules upon which a morphatide performs a preselected or desired function or to which a morphatide binds or associa

Abstract: A method is disclosed herein for monitoring the efficiency of an endonuclease digestion using a plasmid specifically designed for that purpose. The plasmid of the present invention comprises at least two polylinker regions containing a plurality of unique restriction sites distributed so that digestion of the plasmid with any two restriction endonucleases whose sites are represented on the plasmid results in two fragments that are sufficiently different in size from the intact plasmid so as to be readily distinguishable therefrom. To ensure this size differential, the polylinker regions of the plasmid are separated by a spacer segment comprising a restriction site-free nucleic acid sequence that is at least about 15% of the length of the intact plasmid.

Abstract: The present invention provides vectors and methods which improve the efficiency of nucleic acid insertion into circular vectors, which generally facilitate nucleic acid cloning and specifically facilitate the preparation of DNA libraries. In general, the present invention involves separation of the cloning process into two distinct steps: (a) insertion which is done at a high nucleic acid concentration favoring intermolecular joining, and (b) circularization which is performed at a low nucleic acid concentration favoring intramolecular circularization. The present vectors generally have distinct insertion ends and circularization ends which are blocked from covalent joining during the insertion step. Circularization ends contemplated by the present invention include complementary cohesive ends and topoisomerase-linked ends. The present vectors and methods allow minute amounts of nucleic acid inserts to be efficiently cloned.

Abstract: Drug-carrier complexes, drug carriers, pharmaceutical formulations, methods of delivery drugs to an organism or tissue culture, methods of increasing the solubility of a substance, targeted carriers, drug delivery systems and implants are described. The compositions and methods of the invention include forming complexes having reversible associations between nucleotides and drugs. The compositions and methods of the invention can be employed to target drugs to cells, organisms or combinations of cells to treat and to study the underlying mechanisms of diseases, and to test drug candidates.

Abstract: Improved vectors and related materials and methods are disclosed.

Abstract: Methods for the multiplexed detection of the binding of, or interaction between, one or more ligands and target antiligands are provided. Detection involves the release of identifying tags as a consequence of target recognition. The methods include the use of electrophoretic tag probes or e-tag probes, comprising a detection region and a mobility-defining region called the mobility modifier, both linked to a target-binding moiety. In practicing the methods, target antiligands are contacted with a set of e-tag probes and the contacted antiligands are treated with a selected cleaving agent resulting in a mixture of e-tag reporters and uncleaved and/or partially cleaved e-tag probes. The mixture is exposed to a capture agent effective to bind to uncleaved or partially cleaved e-tag probes, followed by electrophoretic separation. In a multiplexed assay, different released e-tag reporters may be separated and detected providing for target identification.

Abstract: The present invention relates to the design, preparation and use of cascade regulatory circuits for amplification of gene expression. The genetic circuit is based on a plurality (e.g., two or more) of regulatory genes organized in a hierarchical order of expression in a genetic construct or constructs, which can be established in a cell, e.g., a gram-negative bacteria, by means of autoreplicative vectors or by chromosomal insertion. In one embodiment, the genetic construct(s) can be stabily maintained in the chromosome without selective pressure, and gene expression induced three orders of magnitude therefrom using economical biodegradable benzoate derivatives.