Abstract: A method of introducing expressible heterologous DNA into Prevotella ruminicola is provided. The method involves conjugal transfer of a shuttle vector comprising the heterologous DNA operatively linked to a promoter functional in P. ruminicola. The invention also provides shuttle vectors for use in the method and P. ruminicola produced by the method. The invention further provides a tetracycline resistance gene of the TetQ class, or fragments thereof that confer tetracycline resistance, and a protein of the TetQ class that provides resistance to tetracycline by protecting ribosomes from tetracycline, or active fragments thereof. Finally, the invention provides a promoter functional in P. ruminicola and an engineered P. ruminicola comprising expressible foreign DNA.

Abstract: An attenuated enterovirus or rhinovirus, suitable for use as a vaccine, has a reversed base pairing in the part, or in a part corresponding to the part, of the 5' non-coding region of the genome of poliovirus type 3 Leon strain shown below: ##STR1## A suitable attenuated poliovirus has the bases G and C at positions 469 and 534 respectively for a type 1 or type 2 poliovirus or at positions 472 and 537 respectively for a type 3 poliovirus.

Abstract: An attenuated enterovirus or rhinovirus, suitable for use as a vaccine, has an attenuating mutation at least at a position which is, or corresponds with, position 471 or 484 of the genome of poliovirus type 3 Leon strain.

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: In a first aspect, the invention involves a reactive support useful for automated synthesis of oligonucleotides. The reactive support comprises a label moiety (e.g. hapten) covalently bonded via a stable bond to a trifunctional spacer. The labeled trifunctional spacer complex is covalently bonded to a solid support via a cleavable bond. One arm of the trifunctional spacer attaches the solid phase; another arm attaches the label; while the third arm provides a hydroxyl group useful for synthesizing a labeled oligonucleotide. Upon synthesis, the cleavable bond is broken, yielding the labeled oligonucleotide. Methods for labeling oligonucleotides and useful kits are also described.

Abstract: Vectors are described that circumvent traditional DNA cloning and subcloning procedures, and that contain a unique DNA cartridge that permits both cloning of DNA directly into DNA sequences present within the cartridge, and in vivo removal and circularization of the cartridge thereby yielding an autonomously replicating structure. Because the DNA cartridge can include a wide variety of functional DNA sequences, the cloned DNA can be subjected to a plethora of molecular biological procedures without having to remove the cloned DNA from the cartridge thereby obviating the need to perform additional subcloning techniques. A particularly useful example of this type of vector is bacteriophage lambda containing the DNA cartridge.

Abstract: An attenuated enterovirus or rhinovirus, suitable for use as a vaccine, has an attenuating mutation at least at a position which is, or corresponds with, position 479 and/or 482 of poliovirus type 3 Leon strain.

Abstract: The subject invention relates to a method referred to as recombination PCR (RPCR). In the method, the polymerase chain reaction is utilized to add double-stranded homologous ends to DNA. These homologous ends undergo recombination in vivo following transfection of host cells. The placement of these homologous ends, by the amplifying primers permits the rapid cloning of the desired mutant or recombinant, with a minimal number of steps and primers.

Abstract: Disclosed are (1) a method of producing D-ribose which comprises cultivating a microorganism belonging to the genus Bacillus having D-ribose producing ability in a medium, the microorganism belonging to the genus Bacillus containing a DNA sequence participating in expression of a gluconate operon which is partly or wholly modified so as to highly express the gluconate operon in the microorganism belonging to the genus Bacillus, accumulating D-ribose, and collecting D-ribose thus obtained; (2) a novel microorganism belonging to the genus Bacillus having D-ribose producing ability transformed with DNA which contains a DNA sequence participating in expression of a gluconate operon which is partly or wholly modified so as to highly express the gluconate operon in the microorganism belonging to the genus Bacillus; (3) novel DNA in which a promoter of a gluconate operon of a microorganism belonging to the genus Bacillus is modified so as to highly express said gluconate operon in the microorganism belonging to the

Abstract: Provided are methods for determining whether a vaccinated animal is uninfected or infected with a virulent wild-type virus. Methods, vaccines and viruses are disclosed that are useful to seratologically distinguish infected from uninfected animals in populations of animals vaccinated with a properly incapacitated virus lacking an antigen of the wild-type virus.

Abstract: The present invention relates to a recombinant DNA molecule which contains the telomere and, optionally, the centromere of a higher eukaryote, particularly a plant, the telomere itself, the centromere itself, a method of producing a polypeptide in a recipient cell which utilizes said recombinant DNA molecule, host cells transformed with said recombinant molecule, and uses for said recombinant molecule.

Abstract: DNA plasmid expression vector, pCMX, enables cDNA expression cloning in mammalian cell culture. This novel expression vector exhibits a marked increase in gene expression when compared to its parental plasmid construction.

Abstract: A cDNA encodes p107; a cell contains recombinant p107-encoding DNA; and substantially all of the cells of a nonhuman mammal contain recombinant p107-encoding DNA. Also, a method for diagnosing a condition of tumorigenicity in a subject, includes the steps of obtaining a tissue sample from the subject and detecting the presence of non wild-type p107-encoding gene in the sample, or detecting the absence of wild-type p107-encoding gene in the sample; or extracting DNA from the sample and detecting the presence of non wild-type p107-encoding gene or the absence of wild-type p107-encoding gene in the DNA. Also, a nucleic acid probe is complementary to a portion of a human mutant p107 gene.

Abstract: A method for determining the developmental stage of a tumor is disclosed. The method comprises the steps of isolating tumor cells, dividing the cells into two samples, transducing one sample with a reporter gene, transducing the other sample with both a reporter gene and a staging gene (wherein the staging gene corresponds to a gene that is differentially expressed in neoplastic and normal cells and is chosen by its ability to be predictive of a certain stage of neoplastic development), analyzing both samples for the level of expression of the reporter gene and comparing the expression level in the samples. In a particularly advantageous embodiment of the invention, the staging gene is JR-ras and the reporter gene is lux.

Abstract: A process for expression of a protein product in Aspergillus is disclosed. The process comprises transforming an Aspergillus strain with a vector system comprising DNA-sequences encoding a promoter including upstream activating sequences derived from an A. niger amylase, a suitable marker for selection of transformants, and a DNA-sequence encoding the desired protein product. The process enables industrial production of many different polypeptides and proteins in Aspergillus, preferably A. niger. Examples of such products are chymosin or prochymosin and other rennets, proteases, lipases and amylases. Also disclosed is an effective promoter for expression of a protein in Aspergillus, preferably Aspergillus niger being derived from a gene encoding an A. niger amylase. The A. niger amylases are the neutral and acid stable .alpha.-amylases and a new amylase not so far described and designated XA amylase. Also disclosed is the novel amylase from A. niger XA amylase.

Abstract: A method of extracting a particular nucleic acid fragment containing a nucleic acid sequence of interest from a nucleic acid or nucleic acids mixture, comprising the steps of:(1) digesting the nucleic acid or nucleic acids mixture with restriction enzymes to obtain a mixture of nucleic acid fragments, said restriction enzymes consisting of (A) two different enzymes capable of producing the particular nucleic acid fragment bearing predetermined and distinct restriction ends on its 5' and 3' terminals and (B) one or more restriction enzymes different from (A), for which the particular nucleic acid fragment contains no relevant restriction sites;(2) preparing two distinct DNA linkers capable of binding to the respective restriction ends of the particular nucleic acid fragment;(3) allowing the linkers to react with the mixture of nucleic acid fragments;(4) subjecting the resulting reaction mixture to the first hybridization with an immobilized probe complementary to one of the linkers;(5) isolating the hybridized

Abstract: The invention relates to a method for selectively cloning exons in which an ukaryotic genomic DNA fragment to be assayed is cloned into a shuttle vector in a cloning site of an intron flanked by exons. Upon expression in eukaryotic cells, exons present in the cloned DNA are spliced to the exons flanking the intron. After amplification of the cDNA of the expressed mRNA with primers specific for the exons in the vector by PCR, selectively cloned exons can be detected due to the size of the PCR product. The invention also relates to vectors used to carry out the method of the invention as well as to exon libraries.

Abstract: Gene therapy involves the transfer and stable insertion of new genetic information into cells. The present invention is directed to safe vectors for gene therapy and thus provides hybrid parvovirus vectors which are capable of site-specific integration into a mammalian chromosome without substantial cytotoxicity, and which direct erythroid cell-specific expression of heterologous genes. The hybrid vector is useful in gene therapy, particularly in the treatment of hemoglobinopathies and other hematopoietic diseases. A method of delivery of constitutive levels of a pharmaceutical product and a method of producing a recombinant protein are also provided.

Abstract: The diagnosis of patients with cancer and other proliferative disorders may be effectively established by the estimation of the expression of prothymosin alpha.

Abstract: The subject invention provides an enzymatically active nonglycosylated, recombinant, human acetylcholinesterase comprising at least one polypeptide characterized by an amino acid sequence which is substantially identical to the amino acid sequence of naturally-occurring human acetylcholinesterase. The subject invention additionally provides an enzymatically active recombinant human acetylcholinesterase comprising at least one polypeptide characterized by an amino acid sequence in which serine is substituted for cys 611 in the sequence of naturally-occurring, human acetylcholinesterase and an enzymatically active recombinant human acetylcholinesterase comprising at least one polypeptide characterized by the presence of a methionine of the N-terminus of the amino acid sequence of naturally-occurring human acetylcholinesterase.