Abstract: DNA strand having an ability in biotechnological production of .alpha.-acetolactate decarboxylase is disclosed. The DNA strand is characterized in that it has a nucleotide sequence coding for a polypeptide whose amino acid sequence is substantially from A to B of FIG. 1 and which has .alpha.-acetolactate decarboxcylase activity. Also disclosed is a yeast which belongs to Saccharomyces cerevisiae and which has been transformed by the DNA strand. The yeast is characterized by the face that its .alpha.-acetolactate producing ability is reduced, and will thus produce an alcoholic liquor such as beer which contains no or little diacetyls which have come from their precursor, namely .alpha.-acetolactate.

Abstract: Novel DNA constructs comprising regulatory regions plus the structural coding region for hepatitis B surface antigen (HBsAg) are disclosed. The regulatory regions employed are responsive to methanol, non-catabolite repressing carbon sources and catabolite repressing carbon sources followed by carbon source starvation. The novel constructs are incorporated into a variety of linear and circular plasmids. Such plasmids are used to transform suitable hosts and ultimately used for the production and isolation of hepatitis B surface antigen in high yields.

Abstract: Surface antigens of swine mycoplasma, such antigens prepared by recombinant DNA methods, a swine mycoplasma vaccine, based on such antigens, methods of treating swine to prevent enzootic pneumonia using that vaccine, and diagnostic tests based on these antigens or antibodies raised against them for detecting the presence of swine mycoplasma infections in swine herds, and DNA sequences that code for such antigens are disclosed.

Abstract: An immunogenicity increasing carrier substance for immunogenic determinants is composed of particles derived from gram-negative cells substantially devoid or depleted of their natural immunogenic determinants, more particularly prepared by stripping the O-antigen and core region off the lipid A by chemical cleavage (acid hydrolysis). These socalled "naked bacteria" are chemically treated with a linking reagent to provide covalently bonded intermediate linking moieties while preserving the adjuvant effect of the particles. The linking moieties have functional groups. These serve for the covalent bonding of selected haptens or antigens comprising the desired immunogenic determinants against which antibodies are to be elicited in living antibody-producing cells.

Abstract: Modified Pseudomonas exotaxins which comprise deletions in at least domain 1A are taught. The toxins exhibit reduced cytotoxicity.

Abstract: A novel hydrophobic polymeric matrix is provided which is suitable for use after implantation in vivo in a subject for the controlled release and delivery of biologically active substances such as drugs, antibiotics, steroids and the like. Alternatively, the matrix can be used outside the body for release of fragrances, pesticides and the like. The implantable matrix comprises a polymeric polyanhydride formulation whose internal anhydride linkages are hydrolytic in nature in varying degrees in accordance with the chemical composition of the backbone, pH and temperature of the environment. As the individual anhydride linkages become hydrolyzed, the matrix erodes predominantly by surface erosion into non-toxic biocompatible degradation products with concomitant release of the biologically active substance.

Abstract: A large DNA cloning system is disclosed which is based on yeast artificial chromosomes. Cloning vectors are disclosed which allow the cloning of large segments of greater than 50 kb of exogenous DNA. The cloning vector comprises DNA sequences of an autonomous replication sequence, a centromere, a selectable yeast marker, two sequences that seed telomere function in vivo, and a cloning site within an interruptible yeast gene for insertion of the exogenous DNA segments.

Abstract: Immunotoxins having improved toxic activity have the composition ##STR1## wherein Toxin-NH is a ribosome-inactivating protein containing no accessible sulfhydryl groups, n is an integer from 1 to 5, m is an integer from 1 to 5, and NH-Antibody is a monoclonal antibody specific to eucaryotic cells or antigens associated therewith. The immunotoxins are made by reacting the Toxin-NH with an iminothiolester salt to form a first conjugate, reacting the NH-Antibody with N-succinimidyl-3-(2-pyridyldithio) propionate to form a second conjugate, and reacting the two conjugates to form the immunotoxin.

Abstract: A method of preparing inocula of low water activity is disclosed. The inocula are prepared by admixing microorganisms with a polymer gel and lowering and maintaining the water activity below 0.3, preferably below 0.1. These inocula are useful in the treatment of plant disease, particularly crown gall and Dutch elm diseases, as well as in the biological control of insects.

Abstract: The whole DNA sequence coding for flagellin of E. coli (hag gene) cloned in phage .lambda. was determined. The hag gene is introduced into pBR322 and the region of the hag gene concerning the antigencity of flagella is lacked, into which linkers are inserted. Where the vector in which foreign DNA is inserted in the linker is introduced in E. coli forming no flagella, the E. coli can form flagella and possesses motility. The flagella comprises flagellin fused with foreign peptide encoded by the foreign DNA. Namely, the foreign peptide is excreted outside of bacteria. This system can be utilized in determination of epitope and preparation of antigen as well as excretion of peptide.

Abstract: Novel DNA sequences which code for the production of Pichia histidinol dehydrogenase, phosphoribosyl-ATP-cyclohydrase and phosphoribosyl-ATP-pyrophosphohydratase are provided. Novel constructs including these sequences, as well as transformed organisms therewith are provided. A method for isolation of functional genes from yeast strains of the genus Pichia is also provided. In addition, process for the integrative transformation of yeast strains of the genus Pichia is provided.

Abstract: Method for the site specific genomic modification of yeasts of the genus Pichia and novel DNA sequences useful therefor are provided. Pichia is transformed with a serially arranged linear DNA fragment comprising first and second insertable DNA fragments, which flank a marker gene. The insertable DNA sequences are homologous to defined portions of the Pichia genome and integrate by recombination at such defined sites. The resulting transformed organism contains the marker gene and any additional DNA sequences which are positioned between the insertable DNA fragments. In addition, the resulting transformed organism has either a disrupted or deleted sequence at the site of the genomic modification. Enhanced production of heterologous gene products is observed when using as the host for expression strains in which the alcohol oxidase gene has been disrupted. Upon disruption of the primary alcohol oxidase gene of Pichia, the existence of a second alcohol oxidase gene in Pichia was also discovered.

Abstract: Methods and compositions are provided for efficient expression of genes in unicellular microoorganisms, particularly yeast. The systems involve an expression system employing transcriptional initiation regions from glycolytic enzymes, particularly a chimeric expression system, having a first region providing for regulatable or constitutive expression, a second region providing for transcriptional initiation, where regions one and two are not found joined together in functional relationship in nature, and optionally a sequence providing for a secretory leader and processing signal, where the expression cassette will be joined to a gene which may be homologous or heterologous to the host. The expression cassette can be used on an extrachromosomal element or integrated into the host genome, whereby continuous expression can be achieved or inducible expression is obtained, by virtue of the presence or absence of an inducer.

Abstract: An aqueous solution containing a physiologically active substance, which is produced by recombinant DNA technique and which has cytotoxic activity against L-M cells and is capable of inducing hemorrhagic necrosis of transplanted Meth A sarcoma in the BALB/c mouse, can be effectively, efficiently purified by column chromatography using a column packed with a dye-bonded crosslinked agarose gel.

Abstract: The present invention relates to plasmids whose hosts can be Escherichia coli and some kinds of yeasts, namely, shuttle vectors, as well as to processes for producing said plasmids. There are provided in the present invention (1) plasmids containing an autonomously replicating sequence of Candida maltosa, Leu 2 gene derived from Saccharomyces cerevisiae and an ampicillin resistance gene and (2) plasmids further containing a tetracycline resistance gene as well as the genes described in (1).The plasmids (shuttle vectors) of the present invention can be utilized as follows. A useful foreign gene is inserted into plasmids of the present invention; using the resulting new plasid, Escherichia coli is transformed and cultured in order to obtain the plasmid in a large amount; and using this plasmid, Saccharomyces cerevisiae or Candida maltosa as a host is allowed to produce useful substances such as hormones and enzymes on a large scale.

Abstract: Process for transforming yeast strains of the genus Pichia is disclosed. Novel yeast strains of the genus Pichia which can be transformed with recombinant DNA material are also disclosed. In addition, a method for isolating functional genes and other functional DNA sequences from yeast strains of the genus Pichia is described.

Abstract: The present invention provides a vector plasmid capable of efficient tumor necrosis factor (TNF) production, a process capable of efficient TNF production in a host transformed with said plasmid and a composition containing the TNF produced by said process.The novel plasmid of the present invention is characterized by having inserted therein a DNA fragment that has a phage-derived promoter region upstream of a structural gene for TNF and in which a DNA fragment containing an E. coli gene-derived transcription termination coding base sequence (terminator) is joined immediately downstream of a base sequence coding for the termination of translation of said structural gene.

Abstract: Expression vectors containing coding sequences under the control of SV40 early and RSV promoters are disclosed as useful in producing proteins in saccharomyces yeasts. Construction of such vectors, and their use in yeast transformations are described.

Abstract: This inverntion concerns a glycoamylase gene cloned into the yeast Saccharomyces cerevisiae, method for cloning such a gene into such yeasts and cloning vehicles containing such a gene, suitable for use in Saccharomyces cerevisiae. Yeast containing a glucoamylase gene are of potential use in the brewing industry.

Abstract: Novel procoagulant proteins are disclosed which comprise the amino acid sequence:A-X-Bwherein region A represents the polypeptide sequence Ala-20 through Arg-759 substantially as shown in Table 1; region B represents the polypeptide sequence Ser-1709 through Tyr-2351 substantially as shown in Table 1; and region X represents a polypeptide sequence comprising up to 949 amino acids substantially duplicative of sequences of amino acids within the sequence SER-760 through Arg-1708 of Table 1, wherein the amino terminus of X is covalently bonded through a peptide bond designated "-" to the carboxy terminus of A, and the carboxy terminus of X is likewise bonded to the amino terminus of B. Methods of making such proteins and their use in pharmaceutical preparations is also disclosed.