A general method for chimerization of monoclonal antibodies by inverse polymerase chain reaction which conserves authentic N-terminal sequences.

Chimerization of antibodies (Ab) by cloning the V (variable) regions encoding the light and heavy chains with degenerate oligodeoxyribonucleotide primers matching to framework region 1 and to the joining regions, leads to Ab with altered amino acids at the N-terminus compared to those of the parental Ab. This is due to N-terminal framework 1 sequences in the expression vectors [Larrick et al., Bio/Technology 7 (1989) 937-938; Le Boeuf et al., Gene (1989) 371-377; Orlandi et al., Proc. Natl. Acad. Sci. USA 86 (1989) 3833-3837]. This might lead to Ab with altered affinity to the antigen due to interaction of framework sequences with complementarity determining regions. Moreover, some V regions may be refractory to cloning by this procedure. Here, we describe a method to circumvent these potential problems. The V regions for both chains of the Ab are cloned by inverse polymerase chain reaction (PCR) with primers matching the known constant region sequences of the Ab. After sequencing, PCR fragments corresponding to the V regions of both chains are inserted in-frame into appropriate expression vectors leading to Ab with unaltered N-terminal sequences after expression in mammalian cells. The procedure is illustrated with an Ab directed against the beta chain of the human interleukin-2 receptor.     

Zeitaufwand für das Ernten und Verstecken von Kiefernsamen bei Dünnschn?bligen Tannenh?hern (Nucifraga caryocatactes macrorhynchos) im Fernen Osten Russlands

Zusammenfassung In einem Vorkommen des Dünnschnäbligen Tannenhähers an der Küste des Ochotskischen Meeres im Fernen Osten Sibiriens wurde das Ernten und Verstecken von Samen aus den Zapfen der Zwergzirbelkiefer (Pinus pumila) untersucht. Der Inhalt von durchschnittlich 2,8 Zapfen, das sind etwa 80 Samen, wurde in der gefüllten Kehltasche transportiert und auf eine Anzahl unter niedriger Zwergstrauchvegetation gelegener Bodenverstecke verteilt. Die Verstecke wurden in annähernd linearer Anordnung ohne Bevorzugung einer bestimmten Himmelsrichtung angelegt. Die Versteckserien enthielten im Median 79, maximal mehr als 120 Samen, das Einzelversteck durchschnittlich 19,6 Samen. Das Ernten und Leeren eines Zapfens geschah im Schnitt innerhalb von 47 s. Für das Verstecken einer Füllung des Kehlsacks benötigten die Vögel ca. 170 s. Für das gesamte Beschaffen und Verstecken eines einzelnen Kiefernsamens errechnet sich ein durchschnittlicher Zeitbedarf von 3,26 s. Nach 20 Tagen war der Zapfenvorrat in der lokalen Kiefernpopulation erschöpft. Jeder Häher hat nach den Hochrechnungen bis zu 100.000 Samen vergraben.

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Harvesting and caching capacities of Thin-billed Nutcrackers in the Russian Far East

Summary At the Ochotskian sea coast Thin-billed Eurasian Nutcrackers (Nucifraga caryocatactes macrorhynchos) harvested seeds of ripe cones of the brush pinePinus pumila in late summer. The mean number of seeds carried in their sublingual pouch was 80, which respresents the harvestable contents of 2.8 cones. These were distributed in an average of 5 caches, exclusively in the soil under low tundra vegetation. Caches were organized in nearly straight lines. Series contained a mean of 82.7 seeds, single caches a mean of 19.6 seeds. Plucking one cone and harvesting its seeds took 47 seconds on average. The caching of a complete pouchful took on average 123.4 seconds. The time invested for harvesting and caching one single seed was calculated at 3.26 seconds. Within three weeks in July, an average individual bird was calculated to have cached a total of up to 100,000 seeds.

     

In-situ investigations on small-scale local and short-term changes of sublittoral macrobenthos in Lübeck Bay (western Baltic Sea)

In-situ studies on sublittoral soft bottom macrofauna (depth: 14–16 m) employing the underwater laboratory (UWL) “Helgoland” were carried out. Sets of samples were compared for small-scale local and short-term changes in species richness, faunal abundance, numerical dominance, diversity, evenness, homogeneity, and similarity. It could be shown that minor differences in sediment quality can cause conspicuous heterogeneity within a small sampling area (diameter: ca. 100 m). Both spatfall and mortality of benthic invertebrates can change the faunal structure within a short period (two months). The degree of change varies between species and thus at stations harbouring different faunal assemblages as well.     

Intracellular and extracellular alkaloids of Penicillium roqueforti

The alkaloid composition of mycelium and culture liquid filtrate of the fungus Penicillium roqueforti IBPM-F-141 was studied. The new metabolite--3,12-dihydroroquefortine, a derivative of roquefortine, the main component of the alkaloid fraction of this culture, has been isolated for the first time. The structure of 3,12-dihydroroquefortine was determined by chemical and physico-chemical methods. In addition to roquefortine and 3,12-dihydroroquefortine, representatives of a new alkaloid group, the clavine alkaloids, e. g. isofumigaclavine A, isofumigaclavine B and festuclavine, were also isolated and identified. The data on the content of these compounds in mycelium and culture medium are presented.