A BamHI family of highly repeated DNA sequences of Nicotiana tabacum

Summary HRS60.1, a monomer unit (184 bp) of a highly repeated nuclear DNA sequence of Nicotiana tabacum, has been cloned and sequenced. Following BamHI digestion of tobacco DNA, Southern hybridization with HRS60.1 revealed a ladder of hybridization bands corresponding to multiples of the basic monomer unit. If the tobacco DNA was digested with restriction endonucleases which have no target site in HRS60.1, the larger part of DNA homologous to HRS60.1 remained as uncleaved relic DNA. These results suggest a tandem arrangement of this DNA repeat unit. Four other clones of tobacco nuclear DNA cross-hybridized with HRS60.1, thus forming a HRS60-family. Sequencing their inserts has shown their strong mutual homology. HRS60-family comprised about 2% of the nuclear genome of N. tabacum. Computer comparisons with other tandem plant-repeated DNA sequences could not detect any other homologous sequence.     

Expression of the threonine operon fromEscherichia coli inBrevibacterium flavum andCorynebacterium glutamicum

Summary The threonine operon fromEscherichia coli was cloned in plasmid pBR322, subcloned into the shuttle vector pCEM300 and the resulting recombinant plasmid was transferred intoBrevibacterium flavum andCorynebacterium glutamicum. The expression ofE. coli threonine genes in these coryneform bacteria was demonstrated by complementing thethrA andthrB mutations and by assaying homoserine dehydrogenase activity.     

Plasmid cloning vectors replicating in Escherichia coli,amino acid-producing coryneform bacteria and Methylobacillus sp.

Summary Four hybrid plasmids were constructed from the cryptic plasmid pAM330 (from Brevibacterium lactofermentum; 4.5 kb) and the broadhost-range plasmid pGV1106 (9.0 kb; Km^r Sm^r) isolated from Escherichia coli. All of them were mobilized from E. coli into the Gram-negative methylotrophic bacterium Methylobacillus sp. and two of these constructs (pCEM300 and pCEM400) were transferred by transformation into B. flavum and Corynebacterium glutamicum. Their kanamycin-resistance determinant coming from Gram-negative hosts was expressed in these Gram-positive bacteria. Both pCEM300 and pCEM400 are very stably maintained in B. flavum and represent suitable vectors for gene cloning in coryneform producers of amino acids.     

Regulation of branched-chain amino acid biosynthesis in Streptomyces fradiae,a producer of tylosin

Synthesis of threonine dehydratase in Streptomyces fradiae was positively influenced by valine and negatively by isoleucine. However, these two amino acids had no effect on the activity of this enzyme. Synthesis of threonine dehydratase in -aminobutyrate resistant mutants of S. fradiae was pronouncedly less sensitive to the positive effect of valine and this change in regulation led to valine overproduction. Synthesis of acetohydroxy acid synthase is regulated in a similar manner to that of threonine dehydratase, however a lower level of expression was detected in -aminobutyrate resistant mutants. And again, no effect of branched-chain amino acids on acetohydroxy acid synthase activity was observed. It follows that in S. fradiae synthesis of threonine dehydratase is the main regulatory mechanism governing production and the mutual ratio of synthesized valine and isoleucine.Abbreviations -AB -aminobutyrate - AHAS acetohydroxy acid synthase - -KB -ketobutyrate - MNNG N-methyl-N-nitro-N-nitrosoguanidine - TD threonine dehydratase - Trans. B. transaminase of branched-chain amino acids - VDH valine dehydrogenase     

Production of milk-clotting enzymes by basidiomycetes

BasidiomycetesPhellinus chrysoloma, Kuehneromyces mutabilis andGanoderma applanatum produce extracellular milk-clotting enzymes. The enzymes are acid proteinases stable at 40°C and within pH 3–5.5. Only the enzyme preparation fromP. chrysoloma exhibits properties comparable with animal chymosin.     

ΔF508 frequency and associated haplotypes near the cystic fibrosis locus in the Yugoslav population

Summary Chromosomes from 19 unrelated Southern Yugoslav families in which cystic fibrosis (CF) occurs were analysed for the presence of the ΔF508 mutation, using polymerase chain reaction amplification followed by dot blot and polyacrylamide gel analysis. Of the 38 CF chromosomes, 15 (39.5%) carry the ΔF508 deletion. Restriction fragment length polymorphism haplotypes for KM19/PstI, XV2c/TaqI and J3.11/PstI marker loci were determined and are compared for a total of 34 N and 37 CF chromosomes.     

Zur Morphologie und Systematik desTrebouxia-Phycobionten im Thallus vonUsnea longissima (Lecanorales)

In the lichen genusUsnea different species ofTrebouxia-phycobionts as well as different haustorial types are known. The isolated and cultivated phycobiont ofUsnea longissima Ach. was studied by light- and electron microscopy and resembles in cytomorphological details the type ofTrebouxia impressa Ahmad. In addition to simple wall-to-wall contacts between the symbiotic components also intraparietal (=intrawall-)haustoria could be observed as the normal interaction type.

Frau Prof. Dr.Elisabeth Tschermak-Woess zu ihrem 70. Geburtstag gewidmet.     

The temporal region and the supramastoid ridge in mesolithic skulls from Padina in the Iron Gate Gorge of the Danube

Studies of the morphological features of the temporal region of mesolithic skulls from Padina in the Iron Gate Gorge of the Danube revealed a very prominent and large supramastoid ridge which is the most striking feature in skulls of both sexes. Mastoid processes were larger in male skulls, but in 25% of the cases there was an overlap between the size of the processes in male and female specimens. The mastoid ridge was prominent in both sexes. The digastric fossa was always well defined in both sexes and in the two thirds of the skull specimens it was deep. The posterior root of the zygoma was prominent in all the skulls, but it was better developed in the male specimens. The parietotemporal suture in both sexes rises above the level of the pterion. There were no morphological or anthropometrical differences between the left and the right side of individual skulls outside the limits of natural asymmetry. All these morphological characteristics of the temporal region may help in racial and sexual diagnosis of the Mesolithic skulls from the Iron Gate Gorge.     

Immobilization of Escheria alcalescens cells with aspartate ammonia-lyase activity

Summary Immobilization of Escherichia alcalescens cells into genu-carrageenan gel for L-aspartic acid production was studied with respect to the optimized preparation of heterogenous biocatalyst /2.5–3.0% genu-carrageenan, 15% biomass, 50–55 °C, tannin added/.     

Morphological differentiation of immobilizedClaviceps paspali mycelium during semi-continuous cultivation

Summary The morphological development ofClaviceps paspali immobilized in Ca-alginate gel was examined. During consecutive reincubations, the immobilized mycelia differentiated into swollen, arthrosporoid-like cells, which never appeared during fermentation of free mycelium. Such differentiation could be connected with the improved, prologed vitality and metabolic activity of the immobilized cells.