Abstract: Promoters capable of ensuring expression in yeast of genes coding for heterologous polypeptides are disclosed, which comprise a DNA fragment or mutants or sub-fragments thereof, said promoter comprising a definite nucleotide sequence commencing with an EcoRI site and terminated by another restriction site. Also disclosed are vector plasmids containing the promoters, transformed yeasts comprising these plasmids, and a process for preparing polypeptides and especially a 1,4-.beta.-N-acetylmuramidase.

Abstract: The invention relates to a strain of Bacillus thuringiensis, GC 91, a sample of which has been deposited under the accession number NCTC 11821, or a derivative or mutant thereof having entomocidal activity against lepidopterous pests. The invention also relates to a process for producing a strain of Bacillus thuringiensis having improved entomocidal properties by combining into a single strain by plasmid transfer the different entomocidal properties of two respective starting strains. The new strains thus produced are useful in entomocidal compositions.

Abstract: Disclosed are discreet functionally translocatable DNA segments, termined Sterol Regulatory Elements (SRE's), which are fused to heterologous structural genes to provided sterol regulatory capability to the thus formed hybrid gene. The hybrid genes respond to sterols by decreasing the production of messenger RNA. The SRE segments contain as their primary functional nucleotide sequence, a 16 bp sequence referred to as direct repeat 2, isolated from the 5' regions of the human LDL receptor gene. DNA segments which include this 16 nucleotide long sequence similarly confer sterol regulatory capability to previously known promoters such as the HSV TK promoter. Also disclosed are discreet sequences which confer positive transcription promotion to heterologous structural genes and promoters without conferring sterol responsivity. Methods are disclosed for employing these genetic control elements in a myriad of embodiments which provide for a fine-tune control of heterologous genes.

Abstract: DNA sequences encoding proteins processable by secretion leaders in recombinant hosts are described. The DNA sequences encode the NH.sub.2 -terminal region of proteins that are cleaved from a secretion leader and may be secreted through the cell membrane and, if present, cell wall in some cases. Proteins encoded by the DNA sequence have an NH.sub.2 -terminal amino acid sequence conforming to a consensus amino acid sequence that is processable by the particular secretion leader. DNA sequences encoding a consensus amino acid sequence processable by the diphterhia toxin secretion leader are disclosed. A novel Pseudomonas exotoxin and CSF-1 having NH.sub.2 -terminal sequences conforming to a consensus sequence are exemplified.

Abstract: The present invention provides a process for stabilizing creatine kinase by disulphide modification, wherein creatine kinase is reacted in any desired sequence(a) with a molar excess of disulphide and/or thiosulphonate and(b) with a molar excess of water-soluble carbohydrate as carrier.

Abstract: Amino acid sequence of N-terminal region of streptococcal acid glycoprotein (SAGP), and antitumor glycoprotein produced by Streptococcus pyogenes Su, was determined, and DNA probes which are complementary to SAGP gene were synthesized.Chromosomal DNA fragment of S.pyogenes Su, which hybridized with those probes was cloned into E.coli, and a restriction map of plasmid DNA harboring SAGP gene was revealed and upon it, DNA sequence or amino acid sequence of SAGP gene was determined.

Abstract: Recombinant plasmids containing the larvicidal delta-endotoxin gene were constructed by inserting HindIII fragments of the Bacillus thuringiensis var. israelensis 72-75 Md plasmid into the Escherichia coli vector pUC12. Two recombinants producing a 27-kdal toxin (pIP173 and pIP174) were indentified by screening clones in an E. coli in vitro transcription-translation system. Both recombinants comprised pUC12 and common 9.7 kb HindIII fragment of the B. thuringiensis plasmid. The 27,340 Da polypeptide synthesized in vito from pIP174 transformed into E. coli JM101 and from B. subtilis 168 and spoOJ87 containing the 1.2 kb TaqI fragment from pIP173 was lethal to mosquito larvae.

Abstract: Human insulin precursors containing the peptide chain B(1-29)-A(1-21) of human insulin and derivatives thereof with a bridging chain connecting the carboxyl terminus of the B(1-29)-chain with the amino terminus of the A(1-21)-chain are prepared by culturing a yeast host transformed with a replicable expression vehicle capable of expressing a DNA-sequence encoding the insulin precursor. The bridging chain is preferably relatively short and contains preferably from 2 to 8 amino acid residues. The bridging chain must not contain two adjacent basic amino acid residues (Lys or Arg) and has one Lys or Arg connected to the amino terminus of the A(1-21)-chain. Human insulin is prepared from the insulin precursors by in vitro conversion.

Abstract: A description is given of a genertic engineering process for the preparation of mesophilic microorganisms which contain a hydantoinase active at elevated temperature, and of DNA sequences which code for this enzyme.

Abstract: A glycoproteinase derived from the culture of Pasteurella haemolytica cleaves carbohydrate antigen markers from glycoproteins forming an integral part of plant and mammal cell membrane without general enzyme digestion of other cell protein. The glycoproteinase is separated from the media containing nutrients which induce Pasteurella haemolytica to produce the glycoproteinase released into the media.

Abstract: Disclosed is a simple method for cultivation of bacteria of the genus Campylobacter with good growth of bacteria which comprises carrying out the cultivation in a sealed container of gas barrier type in which are enclosed a medium inoculated with bacteria and a carbondioxide-generating type oxygen-removing composition packed in a gas permeable packaging material, oxygen concentration and carbon dioxide concentration in the container being adjusted to 0.5-15% and 0.5-22%, respectively within 5 hours from the initiation of the cultivation and these concentrations being kept for at most 72 hours.

Abstract: A purified thermostable enzyme is obtained that has unique characteristics. Preferably the enzyme is isolated from the Thermus aquaticus species and has a molecular weight of about 86,000-90,000 daltons. The thermostable enzyme may be native or recombinant and may be used in a temperature-cycling chain reaction wherein at least one nucleic acid sequence is amplified in quantity from an existing sequence with the aid of selected primers and nucleotide triphosphates. The enzyme is preferably stored in a buffer of non-ionic detergents that lends stability to the enzyme.

Abstract: A process for producing L-tryptophan in which a suitable substrate such as a carbohydrate, indole or anthranilic acid is contacted with Coryneform bacteria, where the Coryneform bacteria bear recombinant DNA constructed by connecting a gene coding for tryptophan synthetase with a plasmid vector capable of proliferating in Coryneform bacteria.

Abstract: Methods of obtaining N-terminal methionine-free proteins are described. The methods employ a novel enzyme, E. coli methionine aminopeptidase either in vitro or in vivo. For in vivo application, plasmid-borne DNA encoding the peptidase is transformed into a bacterial host which produces the desired protein.

Abstract: The cDNA of a new proteinase inhibitor was isolated from a cDNA library of human liver biopsy material using a synthetic Oligonucleotide probe. The cDNA can be expressed by introduction of the cDNA to a microbial host on a DNA vector. Analogs of known proteinase inhibitors can be producing by alteration of the cDNA by substituting codons in the active center of the new proteinase inhibitor.

Abstract: Processes for the preparation of double auxotrophic mutants of Pichia pastoris are provided, as well as novel double auxotrophic mutant strains produced thereby.

Abstract: A method for the preparation of plasmid DNA and the expression products thereof. The method provides cells, enriched in plasmid DNA and expression products thereof, which comprise chromosomal DNA which has been substantially degraded by exposing a plasmid DNA-containing organism to an effective dose of ionizing radiation.