Abstract: Methods for using reporter mycobacteriophage (RM) and p-nitro-.alpha.-acetylamino-.beta.-hydroxy-propiophenone (NAP) to identify TB complex mycobacteria and distinguish these species from MOTT. RM-infected MOTT show little or no reduction in signal when treated with NAP. In contrast, TB complex mycobacteria infected with RM are distinguishable from RM-infected MOTT by a reduction in signal with NAP treatment.

Abstract: A substrate selective for detection of mammalian 5-C-DNA methyltransferase in the presence of bacterial 5-C-DNA methyltransferase, said substrate comprising oligomeric DNA which contains at least one 5-methylcytosine residue, and at least one cytosine or 5-fluorocytosine residue, each of which are followed in linkage to a guanine residue. The invention also includes a method for measuring the presence of mammalian 5-C-DNA methyltransferase which comprises contacting a sample containing 5-C-DNA methyl transferase with the substrate and also includes a method for inhibiting mammalian 5-C-DNA methyltransferase which comprises contacting a sample containing 5-C-DNA methyl transferase with the substrate.

Abstract: An in vitro method for constructing an oligonucleotide concatamer library is revealed. The method has four steps. The first step involves the generation of a primer-bridged circular template oligonucleotide. In the second step, the primer-bridged circular template is used to generate a single-stranded oligonucleotide concatamer by rolling circle replication. In the third step, the single-stranded oligonucleotide concatamer is converted to a double-stranded oligonucleotide concatamer. Finally, the double-stranded oligonucleotide concatamer is cloned or used directly in in vitro assays which allow molecules of interest to be isolated or amplified.

Abstract: An effective method for the production of a gene product employs recombinant DNA vectors comprising a gene encoding a TIF of a eukaryotic translation factor preceeded by a weak eukaryotic promoter, and at least one internal ribosomal entry site region followed by a gene encoding a desired gene product.

Abstract: This application relates to screening methods for identification of antimycotic agents active in mycotic cell translation, the agents identified thereby, and uses of these agents.

Abstract: The present invention relates to compositions for the diagnosis, prevention and treatment of tumor progression. Novel nucleic acid molecules are identified that are expressed at higher levels in benign (e.g., non-malignant) tumor cells compared to malignant tumor cells exhibiting a high metastatic potential. The nucleic acids and cells including these nucleic acids can be used diagnostically or for therapeutic intervention.

Abstract: A method for forming directionally clonable randomly primed cDNA molecules includes the step of priming first strand synthesis using a set of first strand cDNA primers having the sequence 5'-XXNNNNNN-3' where XX is a constant dinucleotide pair across the set and NNNNNN is a random hexanucleotide, the set including primers representing all random hexanucleotides.

Abstract: The present invention provides novel rotavirus reassortants, vaccines employing the novel reassortants and methods for their preparation and administration. One such reassortant contains the gene encoding the v.p.7 neutralization antigen of a human rotavirus. Another reassortant contains the gene encoding the v.p.4 neutralization antigen of a human rotavirus. The remaining genes are provided solely from the bovine rotavirus WC3 strain, or from both the human and bovine strains.

Abstract: Nucleic acid molecule having an RNA substrate cleaving enzymatic activity which cleaves a separate RNA substrate at a cleavage site. The nucleic acid molecule includes an RNA substrate binding portion which base pairs with the RNA substrate only 3' of the cleavage site, and an enzymatic portion (which may include a part or all of the RNA substrate binding portion) having the enzymatic activity. The nucleic acid molecule is able to base pair with the RNA substrate only 3' of the cleavage site, and cause cleavage of the RNA substrate at that cleavage site. The nucleic acid molecule can be either linear or circular. A general method for forming circular RNA in vivo and in vitro is provided.

Abstract: The present invention provides a process for producing substances such as L-tryptophan and L-threonine, which comprises transforming a serine-requiring microorganism belonging to the genus Corynebacterium or Brevibacterium by incorporation of a recombinant plasmid containing a gene which complements the serine-requirement of the host and a gene which relates to the biosynthesis of a desired substance; culturing the obtained transformant in a culture medium; allowing the substance produced by the transformant to accumulate in the culture; and recovering the substance from the culture.

Abstract: A prothrombin time reagent is disclosed for use in a prothrombin time test. The reagent utilizes recombinant human tissue factor, phospholipids of a natural or synthetic origin, a buffer composition and calcium ion. Stabilizers and salts may also be utilized in the reagent. In addition, a method for creating lipid micelles containing tissue factor is also disclosed.

Abstract: Methods for determining the sequence of nucleic acids by cleaving the nueleic acid unilaterally from a first end with an exonuclease activity to sequentially release individual nucleotides, identifying each of the sequentially release nucleotides by mass spectrometry, and determining the sequence of the nueleic acid from the identified nucleotides are disclosed. The method is amenable to multiplexing for simultaneously determining more than one nucleic acid sequence.

Abstract: An computer-based iterative method for analyzing partial gene sequences. Putative gene assemblies are built from partial gene sequences by the method. A series of pre-processing steps prior to assembly allows efficient and accurate assembly of large amounts of partial gene sequences.

Abstract: The invention pertains to a novel method and plasmid system for producing abundant quantities of CRM197 protein, diphtheria toxin or other CRM proteins related to diphtheria toxin, as well as to microorganisms transformed with the novel plasmid. A particularly preferred DNA plasmid, designated pPX 3511, that combines the gene for CRM197 from the nontoxigenic betaphage and the plasmid pNG2-22 is described. The novel plasmid system is capable of transforming strains of Corynebacterium diphtheriae into strains which are capable of expressing high levels of the CRM197 protein without the use of multiple lysogens. The invention provides an elegant means for increasing protein production without having to manipulate the expression vector, such as by increasing the promoter strength, or removing the promoter from iron regulation.

Abstract: A method of selecting transfected cell lines for heterologous protein expression from a population of cells some of which include an endogenous sequence for an amplificable marker and some of which do not include the sequence. The method is based on a minimum amplifiability index which, contrary to current practice, focuses cell lines having relatively low productivity prior to amplification. Transfected cell lines selected with the instant invention produce greater amounts of heterologous protein than cells selected according to the prior art. In one embodiment, the amplifiability index is used to select for further amplification a cell line transfected with a vector containing the coding sequence for human Factor VIII.

Abstract: The present invention provides a process for producing substances such as L-tryptophan and L-threonine, which comprises transforming a serine-requiring microorganism belonging to the genus Corynebacterium or Brevibacterium by incorporation of a recombinant plasmid containing a gene which complements the serine-requirement of the host and a gene which relates to the biosynthesis of a desired substance; culturing the obtained transformant in a culture medium; allowing the substance produced by the transformant to accumulate in the culture; and recovering the substance from the culture.

Abstract: A DNA sequence derived by the mutation from a ColE1-type replication origin is provided, wherein the sequence is characterized in that it includes at least one of the following mutations, which are defined in relation to the wild ColE1 origin: a Guanine to Adenine transversion in position 2976; and a Guanine deletion in position 3057. This sequence increases the number of copies of the plasmids which include it. Plasmids containing the replication origin, and their uses in the amplification of DNA sequences and the production of polypeptides by genetic engineering, are also disclosed.

Abstract: A method of purifying full length synthetic target oligonucleotides from a mixture of oligonucleotides, particularly from a mixture containing truncated or failed sequences. The method involves attaching a short nucleotide sequence complementary to the 5' end of a target oligonucleotide to a solid support. The complementary between the most 5' nucleotides of the target oligonucleotide and the bound oligonucleotide results in hybridization which serves to retain the target oligonucleotide. Truncated or failed sequences lacking 5' sequences complementary to the attached oligonucleotide, fail to hybridize and therefore are not retained. The method makes it possible to purify gram quantities of synthetic deoxyribonucleic acids or ribonucleic acids and sequences which have modifications, such as on the phosphate backbone. The support-bound nucleotide sequences are stable under conditions of purification and therefore can be reused.