Abstract: A process is disclosed for preparing a protein by a eukaryote transformed by multicopy integration of an expression vector into the genome of a yeast, such as Saccharomyces, Hansenula, and Kluyveromyces, or of a mould such as Aspergillus, Rbizopus and Trichoderma, the expression vector containing both an "expressible gene" encoding the protein and a so-called "deficient selection marker meeded for the growth of the yeast or mould in a specific medium", such as the LEU2d, TRP1d or URA3d gene, in combination witha ribosomal DNA sequence, resulting in stable high copy integration of 100-300 copies per cell. This multicopy integration results in an increased production of the desired protein, which can be guar .alpha.-galactosidase, an oxidase or a hydrolytic enzyme such as a lipase.

Abstract: The nucleotide sequence of the P2 gene and the derived amino acid sequence of the P2 protein of Haemophilus influenzae type b are disclosed and the methods used to determine the same. Also disclosed are the methods used to clone and express the P2 gene as well as the purification protocol for the gene and protein. Also disclosed is the synthesis of peptides corresponding to the N-terminal and C-terminal ends of the P2 protein. Also disclosed is the use of the P2 protein as a protective agent for the disease caused by Haemophilus influenzae type b and the use of the protein as a carrier for conjugation with an oligosaccharide derived from Haemophilus to generate a potentially efficacious vaccine against the disease. Also disclosed is the use of P2 peptide-conjugates as immunising agents.

Abstract: The present invention provides compositions and methods for producing a heterologous protein of interest by inserting a copy of a gene encoding the heterologous protein of interest into the chromosome of a host cell, such as E. coli. A chromosomal transfer DNA (a circular, non-self-replicating DNA) is used to integrate the gene encoding the heterologous protein of interest into the host cell chromosome. The chromosomal transfer DNA comprises at least one selectable marker and may optionally include repeated DNA sequences flanking the selectable marker, facilitating chromosomal amplification of the integrated DNA. The gene encoding the protein of interest may be expressed after integration into the chromosome of the host cell; selection for chromosomal amplification may be performed prior to expression of the gene.

Abstract: Recombinant negative-strand viral RNA templates are described which may be used with purified RNA-directed RNA polymerase complex to express heterologous gene products in appropriate host cells and/or to rescue the heterologous gene in virus particles. The RNA templates are prepared by transcription of appropriate DNA sequences with a DNA-directed RNA polymerase. The resulting RNA templates are of the negative-polarity and contain appropriate terminal sequences which enable the viral RNA-synthesizing apparatus to recognize the template. Bicistronic mRNAs can be constructed to permit internal initiation of translation of viral sequences and allow for the expression of foreign protein coding sequences from the regular terminal initiation site, or vice versa.

Abstract: A tetracycline-regulated system which provides autoregulatory, inducible gene expression in cultured cells and transgenic animals is described. In the autoregulatory plasmid pTet-tTAk, a modified tTA gene called tTAk was placed under the control of Tetp. Tetracycline prevents tTA from binding to Tetp, preventing expression of both tTA and luciferase. This negative feedback cycle ensures that little or no tTA is produced in the presence of tetracycline, thereby reducing or eliminating possible toxic effects. When tetracycline is removed, however, this strategy predicts that tiny amounts of tTA protein (which may result from the leakiness of the minimal promoter), will bind to Tet-op and stimulate expression of the tTAk gene. A positive feedforward loop is initiated which in turn leads to higher levels of expression of tTA and thus, luciferase.

Abstract: Recombinantly produced serum albumin is purified in a series of steps, optionally by incubation with an anion-exchange adsorbent, followed by affinity chromatography employing a hydrophobic solid phase and using a water-soluble lipid anion as desorbens in the aqueous phase. This immobile phase comprises a carrier coupled to a 2-mercapto or 2-hydroxy alkanoic acid.

Abstract: A method of preparing biologically active human myeloperoxidase including the steps of preparing a vector for the expression of human myeloperoxidase. The vector includes plasmid pNIV2703. The plasmid includes a HindIII-SnaBI, HindIII-EcoRV or HindIII-HpaI cassette, the cassette including a complete DNA sequence for hMPO as shown in FIG. 1. CHO cells are transformed with the vector. The cells are cultured. The biologically active human myeloperoxidase is recovered.

Abstract: Infection of human fibroblast cells with human cytomegalovirus (HCMV) causes down regulation of cell surface expression of MHC class I. The present invention is directed to a mutant with a 9-kb deletion in the S component of the HCMV genome (including open reading frames IRS1-US9 and US11) which failed to down regulate class I heavy chains. By examining the phenotypes of mutants with smaller deletions with this portion of the HCMV genome, a 7-kb region containing at least 9 open reading frames was shown to contain the genes required for reduction in heavy chain expression. Furthermore, it was determined that two subregions (A and B) of the 7-kb region each contained genes which were sufficient to cause heavy chain down regulation. In subregion B, the US11 gene product is involved. It encodes a endoglycosidase H-sensitive glycoprotein which is intracytoplasmic, similar to the adenovirus type 2 E3-19K glycoprotein which inhibits surface expression of class I heavy chains.

Abstract: Analogs of blood factors which are transiently inactive are useful in treatment of diseases characterized by thrombosis. In addition, modified forms of activated blood factors that generate the active blood factor in serum but have extended half-lives are useful in treating hemophilic conditions. These modified forms of the blood factor may be acylated forms which are slowly deacylated in vivo.

Abstract: The present invention provides a virus comprising a DNA sequence essential for replication of a human cytomegalovirus and at least one foreign DNA sequence adapted for expression in a host. The foreign DNA sequence may encode a human immunodeficiency virus anti-sense mRNA sequence or an antigenic polypeptide, e.g., a malarial surface antigen. Also provided are therapeutic compositions and vaccines which comprise the novel viruses of the present invention.

Abstract: This invention relates to the cloning and expression of autogenes encoding RNA polymerases of T7 and T7-like bacteriophages, in which the RNA polymerase gene is transcribed from a promoter which is recognized by the encoded RNA polymerase. Cloning of T7 autogenes was achieved by reducing the activity of the RNA polymerase sufficiently to permit host cell growth. T7 RNA polymerase activity was controlled by combining two independent methods: lac-repression of the recombinant lac operator-T7 promoter in the autogene and inhibition of the polymerase by T7 lysozyme. Expression systems for producing the RNA polymerases of T7 and other T7-like bacteriophages, and expression systems for producing selected gene products are described, as well as other related materials and methods.

Abstract: Recombinant negative-strand viral RNA templates are described which may be sed with purified RNA-directed RNA polymerase complex to express heterologous gene products in appropriate host cells and/or to rescue the heterologous gene in virus particles. The RNA templates are prepared by transcription of appropriate DNA sequences with a DNA-directed RNA polymerase. The resulting RNA templates are of the negative-polarity and contain appropriate terminal sequences which enable the viral RNA-synthesizing apparatus to recognize the template. Bicistronic mRNAs can be constructed to permit internal initiation of translation of viral sequences and allow for the expression of foreign protein coding sequences from the regular terminal initiation site, or vice versa.

Abstract: A novel chromosomal expression vector based on the bacteriophage Mu is disclosed.

Abstract: This invention provides an infectious retrovirus having inserted between the 5' and 3' long terminal repeat sequences of the retrovirus a nucleic acid encoding an anti-HIV-type specific agent under the control of a pol III promoter. Host cells containing the retroviral vectors of this invention also are provided. Further provided are methods of interfering with or preventing HIV viral replication in a cell infected with HIV or likely to be infected with HIV.

Abstract: The present invention provides recombinant infectious bovine rhinotracheitis (IBR) viruses useful in vaccines to protect bovines from infectious bovine rhinotracheitis and other bovine diseases. The present invention further provides methods for distinguishing an animal vaccinated with the vaccine of the present invention from an animal infected with a naturally-occurring IBR virus. The present invention also provides isolated DNA encoding the gpE glycoprotein of IBR virus and isolated DNA encoding the gpG glycoprotein of IBR virus.

Abstract: Nucleic acid encoding a functional HTLV-III/LAV (HIV-1) protein having trans-activating ability, and expression vectors comprising this nucleic acid are described.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: A WCH-1 cDNA probe specific to mRNA expressing a water channel localized in the kidney collecting tubule and complementary to said mRNA, and the sequence has been identified. The same is obtained bya) subjecting a single-chain cDNA prepared from kidney medullary mRNA of a mammal (rat) to PCR using, as degenerate primers, 5'- (T/C)T(T/C/A/G)AA(T/C)CC(T/C/A/G)GC(T/C/A/G)GT (T/C/A/G)AC- 3' (SEQ ID NO:1) and 5'- AA(T/C/A/G)(G/C)(T/A)(T/C/A/G)C(G/T)(T/C/A/G)GC(T/C/A/G) GG(A/G)TT-3' (SEQ ID NO:2), andb) screening a kidney cDNA library of said mammal using a product of said PCR as a probe. WCH-1 protein molecules constituting said water channel can be produced by Escherichia coli producing protein molecules expressed by WCH-1 gene.

Abstract: Episomal plasmids containing a papovavirus origin of replication and a papovavirus large T antigen mutant form are shown to replicate episomally in human cells, and yield levels of gene expression proportional to their episomal copy number. Candidate oncogenes and anti-oncogenes can be identified by transfecting the episomal plasmids containing a candidate oncogene or anti-oncogene into non-tumorigenic cells and screening for a tumorigenic phenotype.

Abstract: A method for improving the transduction efficiency of retroviral vectors into a host cell wherein the retroviral vectors are incubated with deoxyribonucleoside triphosphates prior to transduction into the host cell is provided.