5S ribosomal gene clusters in wheat: pulsed field gel electrophoresis reveals a high degree of polymorphism

Summary The long-range structure of 5S rRNA gene clusters has been investigated in wheat (Triticum aestivum L.) by means of pulsed field gel electrophoresis. Using aneuploid stocks, 5S rRNA gene clusters were assigned to sites on chromosomes 1B, 1D, 513 and 5D. Cluster sizes were evaluated and the copy number of 5S DNA repeats was estimated at 4700-5200 copies for the short repeating unit (410 bp) and about 3100 copies for the long repeat (500 bp) per haploid genome. A comparison of wheat cultivars revealed extremely high levels of polymorphism in the 5S rRNA gene clusters. With one restriction enzyme digest all varieties tested gave unique banding patterns and, on a per fragment basis, 21-fold more polymorphism was detected among cultivars for 5S DNA compared to standard restriction fragment length polymorphisms (RFLPs) detected with single copy clones. Experiments with aneuploid stocks suggest that the 5S rRNA gene clusters at several chromosomal sites contribute to this polymorphism. A number of previous reports have shown that wheat cultivars are not easily distinguished by isozymes or RFLPs. The high level of variation detected in 5S rRNA gene clusters therefore offers the possibility of a sensitive fingerprinting method for wheat. 5S DNA and other macro-satellite sequences may also serve as hypervariable Mendelian markers for genetic and breeding experiments in wheat.     

SuperSAGE analysis of the <Emphasis Type="Italic">Nicotiana attenuata</Emphasis> transcriptome after fatty acid-amino acid elicitation (FAC): identification of early mediators of insect responses

Background  

Plants trigger and tailor defense responses after perception of the oral secretions (OS) of attacking specialist lepidopteran larvae. Fatty acid-amino acid conjugates (FACs) in the OS of the Manduca sexta larvae are necessary and sufficient to elicit the herbivory-specific responses in Nicotiana attenuata, an annual wild tobacco species. How FACs are perceived and activate signal transduction mechanisms is unknown.     

Die Apogamie bei Aspidium remotum Al. Br.

Ohne ZusammenfassungMit 2 Textabbildungen und Tafel I–X.     

Rapid production of a chimeric antibody-antigen fusion protein based on 2A-peptide cleavage and green fluorescent protein expression in CHO cells

To enable large-scale antibody production, the creation of a stable, high producer cell line is essential. This process often takes longer than 6 months using standard limited dilution techniques and is very labor intensive. The use of a tri-cistronic vector expressing green fluorescent protein (GFP) and both antibody chains, separated by a GT2A peptide sequence, allows expression of all proteins under a single promotor in equimolar ratios. By combining the advantages of 2A peptide cleavage and single cell sorting, a chimeric antibody-antigen fusion protein that contained the variable domains of mouse IgG with a porcine IgA constant domain fused to the FedF antigen could be produced in CHO-K1 cells. After transfection, a strong correlation was found between antibody production and GFP expression (r = 0.69) using image analysis of formed monolayer patches. This enables the rapid selection of GFP-positive clones using automated image analysis for the selection of high producer clones. This vector design allowed the rapid selection of high producer clones within a time-frame of 4 weeks after transfection. The highest producing clone had a specific antibody productivity of 2.32 pg/cell/day. Concentrations of 34 mg/L were obtained using shake-flask batch culture. The produced recombinant antibody showed stable expression, binding and minimal degradation. In the future, this antibody will be assessed for its effectiveness as an oral vaccine antigen.     

The effect of light on the growth rate of intertidal Acropora pulchra (Brook) from Phuket,Thailand, lat. 8°N

Series of apices of Acropora pulchra from an intertidal reef at Phuket, Thailand, were grown at different depths in the sea, and the length growth was monitored at 12–24 h intervals with laser diffraction. The growth rates of actively growing apices were normally distributed and showed a high variability, with an average coefficient of variation of 58%. There was a highly significant difference in average growth rate between adjacent colonies. A significant linear relationship was found between irradiance and length growth, with a saturating level at 300–400 Wm^-2. At 1000 Wm^-2 length growth was significantly reduced. Under normal daylight conditions, day and night growth rates were equal.     

Wingea,a new genus of the Saccharomycetaceae

The yeast species previously described asPichia robertsii v. d. Walt has been transferred to the new genusWingea. The diagnosis for the genus is given.     

Dose-response function for Painter's SCE-model

Summary The replication model for sister chromatid exchange (SCE), when introduced in 1980 by Painter, was claimed to be consistent with the one hit property of SCE. However, the argument offered in favour of the one hit property was based on a defective dose-response function, as shown in this paper, since dose as the independent parameter of any dose-response function was not included in the considerations. This missing part of the model's dose-response function is added and, by using Bessel functions, a formula for the complete dose-response function is presented. A re-examination of the newly derived function shows that, in the model, linearity holds only under certain restricted circumstances.