Abstract: A stable pharmaceutically acceptable formulation containing a pharmaceutically acceptable amount of a protein is disclosed. Also disclosed are associated means and methods for preparing such formulations.

Abstract: This invention relates to a method for the microbial degradation of trichlorethylene by treating trichloroethylene with Pseudomonas mendocina KR-1 or Pseudomonas putida Y2101 or a microorganism host cell that contains a recombinant plasmid. The recombinant plasmid contains toluene monooxygenase genes from Pseudomonas mendocina KR-1. The microogranism host cell containing the recombinant plasmid must have been treated with an inducer of the toluene monooxygenase genes. The method may be applied to the treatment of loci of trichloroethylene chemical waste in water or soil. More particularly, the method may be applied to degrade trichloroethylene as it may be present as a pollutant or contaminant in water, in industrial effluents, in various land areas such as industrial sites, or in various laboratory or commercial installations.

Abstract: A process for isolating and purifying GM-CSF produced from recombinant sources. The process provides for the isolation and purifying of recombinant GM-CSF produced in E. coli.

Abstract: Disclosed and claimed are DNA gene segments, biologically functional plasmids and recombinant plasmids, and microorganism host cells containing such plasmids, all of which contain toluene monooxygenase genes from Pseudomonas mendocina KR-1 and which are useful in a method for the microbial bioconversion of selected phenyl compounds to selected phenolic compounds. In particular, the method is useful for making p-hydroxyphenylacetic acid which is a valuable chemical intermediate in the preparation of certain antibiotics and certain .beta.-adrenergic blocking agents.

Abstract: A method for reducing stored iron and serum iron in mammals is disclosed. In particular, a mammal having an iron overload disorder is administered a therapeutically effective amount of erythropoietin to increase red blood cell production and the mammal is subsequently phlebotomized to remove the excess red blood cells produced.

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, e.g., 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: Novel yeast cell transformation vectors are manufactured and employed in securing expression of exogenous polypeptides in yeast cells. Vectors include promoter/regulator DNA sequences of yeast glyceraldehyde-3-phosphate dehydrogenase gene origins. In an illustrative preferred embodiment, novel immunologically active hepatitis B surface antigen (HBsAg) preparations are isolated from yeast cells transformed with plasmid A.T.C.C. 40053. These HBsAg preparations of yeast origin may be incorporated into vaccine compositions useful in developing immunological responses protective against infection by hepatitis B virus.

Abstract: In processes for recovery of biologically active polypeptides from fermentation cultures of recombinant host organisms, cell death is frequently a prerequisite for isolation processing of the recombinant product outside the fermentation vessel. Disclosed are improved methods for effecting efficient host cell death inside the fermentation vessel through uniformly contacting host cells in culture with microbicidal concentrations of benzyl alcohol. Illustratively, E. coli, B. subtilis, and P. aeruginosa cultures are advantageously treated with from 0.5 to 10.0% (v/v) of benzyl alcohol in the absence of pH or temperature changes within the fermentor.

Abstract: Methods and constructions for controlling the copy number and for maintaining the stability of a plasmid in yeast cells involving inserting into the plasmid a centromere sequence under the transcriptional control of a regulatable promotor. The promoter is not activated when it is desirable to maintain a copy number of one per haploid cell (e.g. during a culture growth phase) but is activated when expression is desired. When the promoter is inactive, the centromere sequence causes the plasmid to behave as a minichromosome, but upon activation of the promoter transcription through the centromere sequence permits an increase in copy number. Multicopy plasmids are selected for by inserting a G418 resistance gene in the plasmid. Medium concentrations of greater than about 400 .mu.g/ml of G418 will select for cells containing multiple copies of the plasmid while lesser concentrations of G418 select for cells containing a single copy of a plasmid containing G418.sup.

Abstract: A class of subtilisin analogs suitable for admixture to cleaning compositions and having improved stability over naturally occurring Bacillus subtilisins are prepared by expressing a modified gene encoding the subtilisin analog in Bacillus subtilis. The subtilisin analogs are characterized as having a modified calcium binding site to improve calcium binding and either an Asn or a Gly replaced in any Asn-Gly sequences present in the subtilisin.

Abstract: Described are rapid and highly efficient procedures for the total synthesis of linear, double stranded DNA sequences in excess of about 200 base pairs in length, which sequences may comprise entire structural genes. Novel sequences are prepared from two or more DNA subunits provided with terminal regions comprising restriction endonuclease cleavage sites facilitating insertion of subunits into a selected vector for purposes of amplification during the course of the total assembly process. The total, finally-assembled sequences include at least one, and preferably two or more, unique restriction endonuclease cleavage site(s) at intermediate positions along the sequence, allowing for easy excision and replacement of subunits and the correspondingly facile preparation of multiple structural analogs of polypeptides coded for by the sequences.

Abstract: Constructions and methods for mutagenesis of nucleic acids involve chemical mutagenesis of a cassette comprising a structural gene linked to a non-functional restorable fragment of a marker gene. Mutants are detected by screening for the presence of the reconstituted marker among the ligation products of the cassette to a vector containing the non-functional restorable remainder of the marker gene. Xylose isomerase mutants, characterized by a change from glu (GAG) to lys (AAG) at amino acid position 262 in the xylA protein of E. coli were obtained by partial marker cassette mutagenesis. These mutants enzymes exhibited twice the rate of isomerization of glucose to fructose exhibited by the wild type.

Abstract: A composition of matter consisting of recombinant human immune interferon having a near-UV circular dichroic spectrum in aqueous solution at neutral pH with positive bands at about 259 nm, 266 nm, 280 nm, and 287 nm, and with shoulders at about 270 nm and 292 nm. Also disclosed is a method for purifying human immune interferon in which proper refolding of the interferon is accomplished by unfolding in a denaturant, such as urea, dilution in ammonium acetate to approximately 0.18 mg/ml of interferon (or less), and dialysis of the solution. The properly folded, purified product which results has a four- to eight-fold greater activity than the aggregate which otherwise results.

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: A probe is attached to a stock support to which is bound a single-stranded polynucleotide by hybridizing both the probe and the bound polynucleotide to a connecting strand with which they are both complementary and then ligating an end of the bound polynucleotide and the probe. After denaturation, the probe remains attached to the support by way of the bound polynucleotide. A target polynucleotide is then hybridized with the support-bound probe and with a labelled probe to provide a detectable hybridization complex.

Abstract: Disclosed are DNA sequences comprising structural genes coding for (1) a polypeptide having the amino acid sequence and properties of urogastrone and for (2) polypeptide analogs thereof which differ in terms of the identity and/or location of one or more amino acids, e.g., [Asp.sup.25 ] and Pro.sup.52, Pro.sup.53 ] analogs of urogastrone. Structural gene sequences may be provided with initial and terminal sequences which facilitate production of discrete protein products by selected host microorganisms as well as for expression by host organisms of fusion proteins, e.g., .beta.-lactamase-urogastrone and .beta.-galactosidase-urogastrone from which the desired products may be isolated.

Abstract: A composition and a method for 5'-labelling polynucleotides undergoing solid phase synthesis wherein a phosphoramidite of an .omega.-hydroxylamine is condensed to a support-bound polynucleotide.

Abstract: A method for the isolation and quantitative detection of a selected single-stranded target polynucleotide from solution. The target polynucleotide is hybridized in solution to a single-stranded mediator polynucleotide, a probe polynucleotide, and an immobilized polynucleotide sequence. The sequence of the mediator polynucleotide comprises a first sequence complementary to a first portion of the target polynucleotide sequence and a second nucleotide sequence complementary to a portion of a single-stranded immobilized polynucleotide sequence. The probe polynucleotide, which carries a detectable label, is complementary to a second portion of the necleotide sequence of the target. The immobilized polynucleotide is immobilized by attachment to a solid support, and, through hybridization to the mediator polynucleotide, functions to immobilize the entire immobilized polynucleotide/target polynucleotide/probe polynucleotide "sandwich".

Abstract: A method for the preparation of a 3' end functionalized polynucleotide is disclosed. An amine-functionalized solid phase support is treated sequentially with an anhydride, then with an .omega.-hydroxylamine. A polynucleotide is chemically synthesized on the treated support and is subsequently cleaved therefrom by hydrolysis of the amide bonds. A polynucleotide having a 3' free primary amine is recovered for use in hybridization assays and other uses.

Abstract: Disclosed are novel circular DNA plasmids useful as vectors in recombinant methods to secure high levels of E.coli expression of exogenous genes. Plasmids of the invention comprise discrete DNA sequences operative to: (1) confer upon the plasmid the capacity for autonomous replication in a host cell; (2) control autonomous plasmid replication in relation to the temperature at which host cell cultures are maintained; (3) stabilize maintenance of the plasmid in host cell populations; (4) direct synthesis of a protein product indicative of plasmid maintenance in a host cell population; (5) provide, in series, a plurality of restriction endonuclease recognition sites, unique to the plasmid and facilitative of exogenous gene DNA sequence insertion; and (6) terminate mRNA transcription of adjacent DNA sequences and situated so as to terminate transcription of exogenous gene sequences inserted within the plasmid at said unique restriction endonuclease restriction sites.