Probing FhuA''-''PhoA fusion proteins for the study of FhuA export into the cell envelope of Escherichia coli K12

Summary The FhuA protein (formerly TonA) is located in the outer membrane of Escherichia coli K12. Fusions between fhuA and phoA genes were constructed. They determined proteins containing a truncated but still active alkaline phosphatase of constant size and a variable FhuA portion which ranged from 11%–90% of the mature FhuA protein. The fusion sites were nearly randomly distributed along the FhuA protein. The FhuA segments directed the secretion of the truncated alkaline phosphatase across the cytoplasmic membrane. The fusion proteins were proteolytically degraded up to the size of alkaline phosphatase and no longer reacted with anti-FhuA antibodies. The fusion proteins were more stable in lon and pep mutants lacking cytoplasmic protease and peptidases, respectively. The larger fusion proteins above a molecular weight of 64000 dalton were predominantly found in the outer membrane fraction. They were degraded by trypsin when cells were converted to spheroplasts so that trypsin gained access to the periplasm. In contrast, FhuA protein in the outer membrane was largely resistant to trypsin. It is concluded that the larger FhuA-PhoA fusion proteins were associated with, but not properly integrated into, the outer membrane.     

Die Weichbodenfauna vor der Elbemündung unter dem Einfluß der Klärschlammverklappung

Macrobenthic fauna and sediment structure were studied at 6 stations from June 1979 to April 1981. The investigations were carried out in the outer estuary of the River Elbe at depths of 15–38 m near a sewage sludge dumping site. The sediment near this area consisted of a high portion of silt. Usually, very few species and small numbers of individuals were found near that dumping ground. During summer 1980, a mass development ofAbra alba and several polychaete species caused an increase in species number which broke down during the following winter. The mass development observed in the study area was apparently not directly caused by the sludge dumping itself, but reflects the natural variability of the community structure. The number of species recorded was significantly correlated with a grain size fraction>31 μm, especially in summer. The seasonal and spatial fluctuation of the benthic community could be well documented by the abundance per species (N/S index). The Shannon-Wiener-Index (H′) and eveness (J) were also useful indicators of community variability. The log-normal distribution did not show the seasonal and the spatial differences of the stations. Possible reasons are discussed. The biomass at the stations near the dumping area was less except during the mass development ofA. alba. The average individual weight (B/N) was shown to be a good indicator of population fluctuations; polychaete biomass was positively correlated to a grain size fraction≦6 μm, that of molluscs to a grain size fraction>31 μm.      

The relationship between short-term memory capacity and EEG power spectral density

Multiplying memory span by mental speed, we obtain the information entropy of short-term memory capacity, which is rate-limiting for cognitive functions and corresponds with EEG power spectral density. The number of EEG harmonics (n = 1, 2,, 9) is identical with memory span, and the eigenvalues of the EEG impulse response are represented by the zero-crossings up to the convolved fundamental, the P300. In analogy to quantum mechanics the brain seems to be an ideal detector simply measuring the energy of wave forms. No matter what the stimulus is and how the brain behaves, the metric of signal and memory can always be understood as a superposition of n states of different energy and their eigenvalues.     

Molecular genetic analysis of 67 patients with duchenne/becker muscular dystrophy

Summary A total of 56 Duchenne muscular dystrophy (DMD) patients and 11 Becker muscular dystrophy (BMD) patients was analyzed by extended multiplex amplification of the DMD/BMD gene; deletions were found in 60% of these patients. The data obtained were used to test the frameshift hypothesis and to compare the distribution of familial versus isolated cases. A significant correlation was found between deletions and isolated cases. Additional experiments were performed in order to determine the deletion breakpoints more precisely. These data are a prerequisite for carrier analysis in the respective families by detection or exclusion of aberrant cDNA fragments derived from ectopic lymphocyte RNA. This diagnostic technique is illustrated by 5 examples.     

Inhibition of microbial growth by interference with siderophore biosynthesis. Oxidation of primary amino groups in aerobactin synthesis by Escherichia coli

Abstract The first step of aerobactin biosynthesis, oxidation of an aliphatic primary amino group to an N -hydroxy-amino compound seems to be involved in the biosynthesis of most of the hydroxamatetype siderophores which are widely distributed among bacteria and fungi. Therefore, the first step of aerobactin biosynthesis, oxidation of lysine to N ⁶-hydroxylysine was studied as a model reaction using a strain of Escherichia coli that contains the first gene aerA of aerobactin synthesis on a multi-copy plasmid and which is lacking the gene for the subsequent step in the pathway. In addition, culture conditions are described which lead to the secretion of N ⁶-hydroxylysine into the medium in amounts that can easily be quantitatively determined by a simple, reliable chemical assay. This assay can be used for screening inhibitors of the oxidation of α-amino groups, which should interfere with the biosynthesis of siderophore hydroxamates and thus should create bacteriostatic conditions.     

Characterization of the genetic system of the xylose-fermenting yeast Pichia stipitis

High mutant frequencies indicated that the wild-type strains of Pichia stipitis are haploid. Sporulation ability of these clones pointed to a homothallic life cycle. Mating was induced by cultivation under nutritionally poor conditions on malt extract medium. Conjugation was followed immediately by sporulation. However, hybrids could be rescued by transferring the nascent zygotes to complete medium before meiosis had started. Under rich nutritional conditions, hybrids were mitotically stable and did not sporulate. The segregation pattern of auxotrophic markers of diploid zygotes indicated regular meiosis, although asci contained preferentially spore dyads. Received: 29 February 1996 / Accepted: 29 March 1996