Bacterial expression of two human aryl sulfotransferases

The effect of replacing a single codon in the N-terminal of human aryl sulfotransferase (HAST) 1 and 3 with one that is more commonly found in E. coli genes was assessed. The pKK233-2 E. coli expression vector was employed and the polymerase chain reaction (PCR) was used to introduce the 5' nucleotide substitution, at the same time maintaining the fidelity of the amino acid sequence. The data indicates that this change had a minimal effect on protein production, subcellular localization or, in the case of HAST3, catalytic activity. In general, the pKK233-2 E. coli vector has been less than optimal for expressing human sulfotransferase cDNAs. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved

Small-scale spatial variability of selected soil biological properties

A strategy for sampling soil from intact monolith lysimeters was established based on measurements of spatial heterogeneity within the lysimeter area. This was part of an ongoing study to determine relationships between soil microbial diversity and nutrient loss by leaching. The sampling protocol had to allow for collection of soil on a regular basis (as opposed to destructive sampling) and ensure high spatial independence of subsamples. On each of two sites (one developed under organic crop management, the other under conventional crop management), ten 15-cm soil cores (sampling points) were taken from three areas (replicates) of 50-cm-diameter (lysimeter surface area) and separately analysed for biotic (microbial biomass carbon and nitrogen; arginine deaminase activity) and abiotic (total carbon and nitrogen) soil properties. The data was tested for variability, expressed as coefficient of variance (biotic and abiotic), and spatial heterogeneity using geostatistics (biotic properties). The biotic soil properties showed significant differences among sampling points, whereas the abiotic parameters were useful in differentiating on a larger scale, i.e. between sites. For all soil properties tested, the differences among the replicates were smaller than those between sites or among points indicating that, in the main experiment, all treatments can be sampled following the same pattern.Geostatistical analysis and fitting of an exponential model showed that a spatial structure exists in the biotic soil properties and that the samples are independent beyond separation distances of 25-30 cm. A revised sampling pattern consisting of 11 samples per lysimeter is described