Abstract: The invention relates to a process for the preparation of a fermentation broth containing coenzyme B.sub.12, with a new, anaerobic, mesophilic, methane-producing mixed micropopulation, under anaerobic, septic conditions, using a new broth containing methanol, precursors and partially known nutrients,In the new process according to the invention the improvement consists in(a) removing 5 to 15 volume percent of the inoculum fermentation broth containing the anaerobic, mesophilic, methan-producing new mixed micropopulation and replacing same with an equal volume of a broth containing cornsteep liquor hydrolysate and/or corn slops of hydrolysate thereof and other known nutrients, the number and concentration of which has been reduced, for 6 to 8 days, and if desired, further manufacturing the fermentation broth removed,(b) after reaching an assimilation velocity of at least 0.15 to 0.2 g methanol/lit.

Abstract: This invention provides a method for high frequency plant regeneration from somatic stem donor tissue of field grown Zea diploperennis, a diploid, perennial corn ancestor with high tillering capacity. This species is used as a parent in a maize improvement strategy to transfer the unique traits of high tillering and plantlet regeneration capacity into cultivated corn. After 3-4 subcultures of cultured somatic tissues on a primary medium, small callus fragments are transferred to secondary medium devoid of the auxin, 2,4-D. After a few days, numerous shoots regenerate and develop into normal plantlets which are then separated and transferred to a tertiary medium for root development. The selection of somaclonal variants form cultured somatic cells of interspecific hybrids between corn and teosinte are used for the synthesis of unique breeding lines suited for development of improved corn varieties. A protocol for gene transfer employing recombinant DNA techniques is also described.

Abstract: Vectors are disclosed for cloning any foreign DNA in prokaryotic and eukaryotic organisms, which vectors comprise a restriction site bank containing a large number of unique restriction sites. The disclosed vectors permit the cloning of large DNA fragments and complete genes without the risk of unwanted DNA cleavage by restriction enzymes. Also disclosed are methods for preparing the aforementioned vectors and processes using the vectors for the cloning of DNA, for the production of polypeptides, and the like.

Abstract: The invention relates to yeasts of the genus Kluyveromyces, in particular K. lactis, acting as hosts for the expression of preprothaumatin or its various allelic and modified forms or their maturation forms, which yeasts contain an rDNA plasmid comprising a structural gene encoding for a thaumatin-like protein, an autonomous replicating sequence derived from K. lactis (KARS), a yeast regulon derived from the GAPDH-genes of S. cerevisiae, and a selection marker, e.g. the trp-1 or the lac-4 gene from S. cerevisiae and K. lactis, respectively. Optionally the plasmid also contains a transcription terminator from a yeast. Further the preparation of such yeasts and the preparation of thaumatin-like proteins with such yeasts and the proteins so obtained are claimed. The yeast described give a better expression than obtained with E. coli as described in EP-PA (A2) 0 054 330 and 0 054 331, published on June 23, 1982.

Abstract: A method for improving the yield of heterologous protein such as IFN-.alpha., IFN-.beta., and IL-2, produced by recombinant bacteria by supplementing the nutrient medium in which the bacteria are grown with a water soluble alcohol and/or amino acid mixture during the terminal phase of the cultivation.

Abstract: This invention provides novel mutant strains of microorganisms (e.g., Pseudomonas putida Biotype A) which are capable of converting substrates such as toluene, p-xylene, catechol and 4-methylcatechol to 2-hydroxymuconic semialdehyde or substituted analog of 2-hydroxymuconic semialdehyde quantitatively by the meta (catechol 2,3-oxygenase) pathway.No active 2-hydroxymuconic semialdehyde-metabolizing enzymes are induced in the microorganism, thereby permitting a 2-hydroxymuconic semialdehyde type metabolite to be produced and accumulated in a bioconversion medium containing the microorganism.

Abstract: The present invention discloses novel selectable recombinant DNA cloning vectors for use in Streptomyces, E. coli and related organisms. The invention further comprises transformants of the aforementioned vectors.

Abstract: A bacterial product is made by transforming a temperature sensitive lysogen with a DNA molecule which codes, directly or indirectly, for the product, culturing the transformant under permissive conditions and externalizing the product by raising the temperature to induce phage encoded functions.

Abstract: A method and apparatus for positive selection of viable cells based upon differing chemical or physical properties or dynamic processes using a focussed radiant energy beam, such as a laser beam (11) to kill unwanted cells is described. The apparatus includes a microscope (14) with a photomultiplier (27) which is used to selectively detect light from certain cells produced by an attenuated laser beam irradiating the cells for identifying the cells to be killed or saved. Based upon identification, selected cells to be killed are exposed to the unattenuated laser beam.

Abstract: A special description is given of plasmids pSM15, pSM16 and pSM17, which are hybrid plasmids derived by coupling pUB110 and modified pE194. The described plasmids are cloning vectors in Bacillus subtilis and are characterised by a marker of resistance to Kanamycin and an EcoRI site located near the end of the ribosome recognition sequence; they can also be used to induce the coded heterologous protein from cloned DNA with sub-inhibiting doses of erythromycin.These plasmids are prepared via the formation of plasmids pSM4, pSM5 or pSM6, which can be expressed either in B. subtilis or in E. coli.

Abstract: A method and apparatus for position selection of viable cells based upon differing chemical or physical properties or dynamic processes using a focussed radiant energy beam, such as a laser beam (11) to kill unwanted cells or to isolate wanted cells is described. The apparatus includes microscope means (14) and cell detection means (27) for identifying the cells to be killed or saved and attenuator means (12) for modifying the beam to prevent destruction of cells to be saved.

Abstract: A plasmid pOA15 in isolated form having the property of conferring pock-forming ability on Streptomyces griseus (ATCC 10137) and a molecular length of about 10 kb, and showing such sensitivity that(a) it has one restriction site recognized by BamHI,(b) it has one restriction site recognized by EcoRI,(c) it has 7 restriction sites recognized by SalI, and(d) it is not cleaved by BglII, HindIII, KpnI, PstI and XbaI,and a biologically pure culture of a microorganism containing the above plasmid.

Abstract: A process is defined for providing plants of a nonhalophytic crop wherein the resulting plants have an enhanced ability to grow successfully in a soil comprising a normally deleterious concentration of a plurality of inorganic salts. Tissue culturing of a large quantity of plant cells initially is carried out under conditions wherein substantial disorganized cell growth takes place which is accompanied by genetic variation (e.g., via somaclonal variation), and the resulting cells subsequently are tissue cultured in a medium which is substantially lacking in a hormone component comprising a plurality of inorganic salts which are provided in a concentration which is normally injurious to cells of the crop wherein a substantial quantity of cells are killed and at least one cell survives and multiplies which has undergone genetic variation wherein an atypical functioning single gene is present which is dominant for increased tolerance to the totality of the inorganic salts present.

Abstract: Provided is a method of obtaining endomycorrhizian fungi with vesicles and arbuscules in vitro. The method comprises producing dicotyledone roots which have been genetically converted by inserting genes of root inducing or tumor inducing plasmid into the genome of dicotyledone roots, and then inoculating the converted roots with endomycorrhiza spores. Application of the present method can be very useful in agriculture and horticulture.

Abstract: The present invention provides human Interleukin 2-encording messenger RNA which is free of human cells, a method of producing the same from the cells which are capable of producing human Interleukin 2 and a method of producing human Interleukin 2 using the same in an in vitro protein synthesis system.