Construction and Characterization of a 10-Genome Equivalent Yeast Artificial Chromosome Library for the Laboratory Rat,Rattus norvegicus

Increasing attention has been focused in recent years on the rat as a model organism for genetic studies, in particular for the investigation of complex traits, but progress has been limited by the lack of availability of large-insert genomic libraries. Here, we report the construction and characterization of an arrayed yeast artificial chromosome (YAC) library for the rat genome containing approximately 40,000 clones in the AB1380 host using the pCGS966 vector. An average size of 736 kb was estimated from 166 randomly chosen clones; thus the library provides 10-fold coverage of the genome, with a 99.99% probability of containing a unique sequence. Eight of 39 YACs analyzed by fluorescencein situhybridization were found to be chimeric, indicating a proportion of about 20–30% of chimeric clones. The library was spotted on high-density filters to allow the identification of YAC clones by hybridization and was pooled using a 3-dimensional scheme for screening by PCR. Among 48 probes used to screen the library, an average of 9.3 positive clones were found, consistent with the calculated 10-fold genomic coverage of the library. This YAC library represents the first large-insert genomic library for the rat. It will be made available to the research community at large as an important new resource for complex genome analysis in this species.     

Die durch Röntgenstrahlen induzierte elektrische Leitfähigkeit in dem PVC-Kunststoff „Trovidur“

Zusammenfassung An dem Kunststoff Trovidur, einem Erzeugnis auf der Grundlage von Polyvinylchlorid, wurde die Änderung der Leitfähigkeit bei der Bestrahlung mit hohen Dosisleistungen untersucht. Als Strahlenquelle diente eine im Max Planck-Institut für Biophysik in Frankfurt a. M. konstruierte Hochleistungsröntgenanlage. Während der Bestrahlung mit Dosisleistungen von 3,5.10³ bis 3,8.10⁵ rad/min konnte zwischen dem strahleninduzierten Stromi und der DosisleistungL eine Abhängigkeit gemäß der Beziehung iL^0,75+-0,02 festgestellt werden. Die spezifische Leitfähigkeit nahm bei einer Bestrahlung mit 3,8·10⁵ rad/min um den Faktor 1,3·10⁴ von 8·10^–19 Ohm^–1cm^–1 auf 1·10^–14 Ohm^–1cm^–1 zu. Das zeitliche Abklingen des strahleninduzierten Stromes nach Beendigung der Bestrahlung konnte durch eine Überlagerung zweier hyperbolischer Funktionen mit den Zeitkonstanten 0,1 bzw. 38 min angenähert werden. Zur Deutung der Ergebnisse sind Modellvorstellungen und theoretische Überlegungen vonRose sowie vonBroser undBroser-Warminsky herangezogen worden. Wesentlich ist im vorliegenden Fall die Annahme einer besonderen (exponentiellen) Haftstellen verteilung.Gewidmet Herrn Prof. Dr. Dr. h. c. Dr. h. c. B.Rajewsky zum 25jährigen Bestehen des Max Planck-Instituts für BiophysikHerrn Professor Dr. B.Rajewsky möchte ich für die Anregung zu dieser Arbeit und für deren stete Förderung meinen herzlichen Dank aussprechen. Den Herren Dr. K.Heuss und Dr. A. P.Lotz bin ich für gute Ratschläge, wertvolle Hilfe und Diskussionen zu großem Dank verpflichtet. Dank schulde ich auch Herrn G.Manthey für seine tatkräftige Unterstützung bei den Bestrahlungen.     

The role of prophenoloxidase activation in non-self recognition and phagocytosis by insect blood cells

Experiments indicate that the prophenoloxidase activating system, which is responsible for melanin production, is also involved in immunorecognition in insects. Using haemocyte monolayer preparations of Blaberus craniifer, Galleria mellonella and Leucophae maderae, it was shown that laminarin, a β 1,3-glucan extracted from fungal cell walls and an activator of the prophenoloxidase system, enhanced the phagocytosis of test bacteria.Scanning electron microscopy of haemocyte monolayers showed that incubation of test bacteria with laminarin significantly increased the number of microorganisms attached to both the plasmatocytes and the granular cells. Furthermore with the granular cells, these bacteria became entrapped in an amorphous matrix. This material probably consists of the “sticky” proteins previously reported to be produced by crustacean haemocytes following prophenoloxidase activation. Pretreatment of haemocytes with laminarin abolished the stimulatory effect on ingestion, indicating that these “sticky” proteins are opsonic, since they would have been discharged from the haemocytes onto the glass monolayer leaving few molecules available for subsequent coating of the test particles.Preliminary biochemical studies on the G. mellonella prophenoloxidase system demonstrated that it was activated by trypsin, laminarin and laminarin G, a highly purified β 1,3-glucan, but not by dextran. Serine protease activities were also enhanced by adding laminarin to a haemocyte lysate supernatant, suggesting that the stimulatory mechanism may involve the proteolytic activity of such enzymes.     

A Diazo Coupling Method for the Electron Microscopic Localization of Cholinesterase

The presence of cholinesterase at the myoneural junction of intercostal muscle has been demonstrated in both light and electron microscopic preparations. A new simultaneous diazo coupling technique using α-naphthyl acetate as substrate and "hexazonium pararosanilin" as coupler has been applied to cold formalin-fixed tissues. After postfixation in buffered osmium tetroxide the sites of esterase activity are faithfully demonstrated at a high level of resolution. The details of cholin-esterase distribution and some technical aspects of the procedure are discussed.     

Strong land‐use effects on the dispersal patterns of adult stream insects: implications for transfers of aquatic subsidies to terrestrial consumers

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