Abstract: A chemically-synthesized oligonucleotide composing a portion of the nucleotide sequence of the human IL-2 is employed as a probe to isolate the gene coding for human IL-2. The human IL-2 gene is selected from a cDNA library prepared from RNA produced by mitogen-stimulated Jurkat cells. Double-stranded cDNA is prepared from polyadenylated RNA extracted from bovine cells thought to produce interleukin-2. Such cDNA is inserted within a plasmid vector and the recombinant plasmid employed to transform hosts. Plasmid DNA, prepared from pools of the transformed hosts, is hybridized with a probe composed of a large portion of the coding sequence of the human IL-2 gene. Pools of host cells that provide signal to the human cDNA probe are identified, subdivided, and rescreened until a single positive colony is identified. Bovine plasmid cDNA is prepared from this colony, and the bIL-2 gene is sequenced.

Abstract: The present invention discloses several DNA mutagenesis processes using a DNA template containing several uracil residues in place of thymine, which can be applied without selection techniques to produce altered DNA sequences with approximately 10-fold greater efficiency than current methods of site-specific mutagenesis.This template has relatively normal coding potential in the in vitro reactions typical of standard site-directed mutagenesis protocols but is not biologically active upon transfection into a wild type (i.e., ung.sup.+) E. coli host cell. Expression of a desired change, present in the newly synthesized non-uracil-containing covalently closed circular complementary strand, is thus favored. The procedure has been applied to mutations introduced via both obligonucleotides and error-prone polymerization. The inclusion of two additional simple treatment steps before transfection results in a site-specific mutation frequency approaching 100%.

Abstract: Recombinant herpes viruses useful as effective ingredients in vaccines against both virulent HSV-1 and HSV-2 are disclosed. Methods of preparing the recombinant viruses, vaccines incorporating the viruses, and methods of immunizing a human host by inoculation with the vaccines are also disclosed.The genomes of the recombinant viruses each comprise a mutant of the HSV-1 genome from which a portion thereof responsible for neurovirulence yet nonessential for growth is deleted. A gene from the HSV-2 genome responsible for coding an immunity-inducing glycoprotein is inserted in the mutant genome between the end points of deletion therein.

Abstract: The DNA coding for human recombinant growth hormone releasing factor (GRF) cloned into a bacterial plasmid expression vector by assembling in one step selected synthetic DNA fragments.

Abstract: The present invention presents a DNA containing a synthetic gene for expression of human epidermal cell growth factor of the DNA sequence; ##STR1## The adoption of the DNA of the present invention improves the stability of mRNA, the translation efficiency and the influence of the high-dimensional structure which is determined by the mRNA sequence, and made it possible to produce hEGF efficiently.

Abstract: Disclosed herein is a method for amplifying and expressing transferred genes by temperature induction.

Abstract: Regulation of eucaryotic gene expression is controlled by procaryotic peptides. The peptides recognize specific DNA sequences present in the gene, which may be derived from procaryotic genes, and either activate or repress gene transcription. Hybrid procaryotic peptides may be used containing both repressor and activator peptides.

Abstract: The invention concerns the intercistronic DNA sequence of the atp operon of Escherichia coli for the subunit c of the ATP synthase, a DNA structure and an expression vector which are characterized by the said intercistronic DNA sequence, and a process for the production of proteins using the same. The DNA sequence enhances the rate of synthesis of the proteins.

Abstract: Disclosed are cultured mouse cells with amplified adenosine deaminase genes which greatly overproduce adenosine deaminase. The cells were isolated by growth in media containing alanosine, uridine, and a cytotoxic concentration of adenosine. Maximum enhancement of adenosine deaminase activity was achieved by also adding deoxycoformycin to the media and then serially increasing the deoxycoformycin concentration at 4 to 6 week intervals. Also disclosed is a cDNA homologous to mouse adenosine deaminase mRNA. The cDNA was produced by synthesis against RNA extracted from the cells containing amplified adenosine deaminase genes and then inserted into a bacterial plasmid. Multiple copies of the cDNA were produced by replication of the bacteria into which the plasmids were inserted.

Abstract: A process and means for increasing the production of protein translated from eukaryotic messenger ribonucleic acid (mRNA) comprising transferring a regulatory nucleotide sequence from a viral coat protein mRNA to the 5' terminus of a gene or complementary deoxyribonucleic acid (cDNA) encoding the protein to be produced to form a chimeric DNA sequence. The regulatory DNA sequence are generated de novo using genetic engineering procedures to produce synthetic double-stranded oligonucleotides representing the regulatory viral sequence. Nucleotide sequences which encode a regulatory sequence or structure conferring enhanced competitive activity and increased rate of translation upon the chimeric DNA sequences include the nucleotide sequence preceding the initiator AUG codon at the 5' terminus of coat protein messenger RNA from alfalfa mosaic virus, brome mosaic virus, black beetle virus, turnip yellow mosaic virus, and satellite tobacco necrosis virus.

Abstract: A gene of varicella-zoster virus (VZV) which encodes immunogenic outer surface viral proteins has been identified by DNA sequence analysis. This gene can hybrid select messenger RNA which encodes and expresses a protein which reacts with human convalescent zoster sera and with polyclonal monospecific antisera which neutralize viral infectivity. These proteins are useful for the preparation of a vaccine for VZV.

Abstract: Disclosed are novel polypeptides possessing part or all of the primary structural conformation and one or more of the biological properties of a mammalian (e.g., human) pluripotent granulocyte colony-stimulating factor ("hpG-CSF") which are characterized in preferred forms by being the product of procaryotic or eucaryotic host expression of an exogenous DNA sequence. Sequences coding for part or all of the sequence of amino acid residues of hpG-CSF or for analogs thereof may be incorporated into autonomously replicating plasmid or viral vectors employed to transform or transfect suitable procaryotic or eucaryotic host cells such as bacteria, yeast or vertebrate cells in culture. Products of expression of the DNA sequences display, i.e., the physical and immunological properties and in vitro biological activities of isolates of hpG-CSF derived from natural sources. Disclosed also are chemically synthesized polypeptides sharing the biochemical and immunological properties of hpG-CSF.

Abstract: The present invention provides a method for site-specific cleavage of double stranded DNA at sequences not less than eight base pairs long, comprising methylating he DNA with a sequence-specific methylase capable of recognizing and methylating a first sequence of the DNA, thereby generating a second sequence of the DNA capable of being recognized by a site-specific endonuclease, the first and second sequences having an overlapping part thereof; the length of the combined methylase and endonuclease recognition sites being not less than eight base pairs long, and cleaving the methylated DNA by treatment with the site-specific endonuclease. This method is useful for increasing the selectivity of cutting DNA stands using restriction endonucleases, thereby permitting the isolation of large DNA fragments and the generation of unique sites in DNA fragments, e.g., cloning vectors.

Abstract: Plasmids useful as cloning vectors carrying an inserted regulatable promoter, transcription from the promoter regulating plasmid replication. Increased transcription leads to a substantially increased or uncontrolled plasmid copy number when host microorganisms containing the plasmid are cultivated under conditions securing such increased transcription. The regulatable promoter is preferably .lambda. P.sub.R, the activity of which is controlled by a temperature-sensitive .lambda. cl repressor. To obtain gene products of the plasmids, host microorganisms to which such plasmids have been transformed are desirably cultivated at about 30.degree. C. to secure a constant, low plasmid copy number during the seeding and multiplication stages of the microorganisms, after which the temperature is preferably shifted to about 36.degree.-42.degree. C. to obtain an uncontrolled plasmid copy number and an amplified amount of gene product.

Abstract: Methods and vectors for high level expression of genes in bacteria are disclosed. A terminal mRNA sequence from a gene coding for a stable bacterial protein mRNA is ligated to a gene coding for the desired protein adjacent the translation termination codon of the gene. The gene for the desired protein and the terminal mRNA sequence are situated in an expression vector in which the gene is operably linked to a transcription promoter.

Abstract: This invention concerns recombinant RNA molecules comprising a recognition sequence for the binding of an RNA-directed RNA polymerase, a sequence for the initiation of product strand synthesis and a heterologous sequence of interest inserted at a specific site in the internal region of the recombinant molecule. Such recombinant RNA molecules are capable of serving as a template for the synthesis of complementary single-stranded molecules by RNA-directed RNA polymerase. The product molecules so formed are also capable of serving as a template for the synthesis of additional copies of the original recombinant RNA molecule. In a preferred embodiment of the invention Q.beta. replicase is used as the RNA-directed RNA polymerase.

Abstract: High yields of active Factor IX are produced by culturing a CHO cell line transfected with chromosomally-integrated Factor IX cDNA in medium to which vitamin K is added.

Abstract: This invention describes the discovery of a novel phenomena in retrovirus transcription, namely transcriptional trans-activation. Described herein are novel trans-acting factors which may be employed to enhance the production of heterologous genes. Described is a novel trans-acting directing gene, designated herein as the "luk" gene and the 35,000 to 45,000, more specifically about 42,000 dalton molecular weight protein encoded thereby.The present invention demonstrates the LTR elements of HTLV can function as transcriptional promoters for heterologous genes on both unintegrated and integrated DNA. In general, the HTLV-1 LTR is a stronger promoter than is the HLTV-II LTR in its requirements for cellular and/or viral trans-acting factors in order to function efficiently. HTLV infection results in the production of trans-acting factors that dramatically increase the rate of HTLV LTR-promoted transcription.

Abstract: The present invention relates to pseudorabies virus mutants containing deletion and/or insertion mutations in a major viral glycoprotein gene, such that no antigenic polypeptides encoded by the viral gene are produced. As a result, animals vaccinated with such do not develop antibodies to the viral glycoprotein and can be distinguished from animals infected with pseudorabies virus field strains and known pseudorabies virus vaccine strains. The present invention also relates to vaccines for pseudorabies disease containing the same, methods for production of the same and methods for use of the same.

Abstract: A novel recombinant DNA containing the base sequence shown in FIG. 1 or a portion thereof which exhibits promoter activity. The base sequence exhibiting a potent promoter activity is obtained from the chromosomal DNA of strain Bacillus by, for example, cloning with a cloning vector. By growing a transformant of Bacillus transformed with the recombinant DNA carrying a peptide encoding nucleotide, the desired peptide may be produced.