A Novel Conductometric Urea Biosensor with Improved Analytical Characteristic Based on Recombinant Urease Adsorbed on Nanoparticle of Silicalite

Development of a conductometric biosensor for the urea detection has been reported. It was created using a non-typical method of the recombinant urease immobilization via adsorption on nanoporous particles of silicalite. It should be noted that this biosensor has a number of advantages, such as simple and fast performance, the absence of toxic compounds during biosensor preparation, and high reproducibility (RSD = 5.1 %). The linear range of urea determination by using the biosensor was 0.05–15 mM, and a lower limit of urea detection was 20 μM. The bioselective element was found to be stable for 19 days. The characteristics of recombinant urease-based biomembranes, such as dependence of responses on the protein and ion concentrations, were investigated. It is shown that the developed biosensor can be successfully used for the urea analysis during renal dialysis

Parallel stranded duplex DNA.

Three linear 21-nt oligonucleotides (C2, C3, C7) have been synthesized with different sequences of A and T residues. One pairwise combination, (C3, C7), hybridizes to form a conventional antiparallel duplex (aps-C3.C7), whereas the pair C2, C3 forms a duplex (ps-C2.C3) in which the two strands are in a parallel orientation and the A.T base-pairs in a reverse Watson-Crick configuration. The existence of the novel ps helical structure was established from the following criteria: (i) The electrophoretic mobilities of the ps and aps duplexes in native and denaturing polyacrylamide gels are similar. (ii) The ps duplex is not a substrate for T4 DNA ligase. (iii) Salt-dependent thermal transitions are observed for the two duplexes, but the melting temperatures of the ps molecules are 15 degrees C lower. (iv) The ultraviolet absorption and circular dichroism spectra of the ps duplex are indicative of a base-paired structure, but differ systematically from that of the aps helix. (v) Based on fluorescent measurements, the bis-benzimidazole drug BBI-258 shows a lower affinity for the ps compared to the aps duplex, whereas the opposite preference holds for the intercalator ethidium bromide. We conclude from the present study that parallel stranded DNA is a stable conformation which can arise by interaction between two conventional strands with appropriate sequence homology

Identification of sulfate-reducing bacteria using 16S rRNA binding oligonucleotide probes

Fluorescent-dye-conjugated oligonucleotides were used as "phylogenetic" probes to identify sulfate reducing bacteria (SRB) in a photosynthetic biofilm. Two samples of biofilm from a trickling filter in a waste-water treatment plant were incubated for 48 hr's; one in darkness, the other in daylight. Chemical gradients of (02, S2-, and pH) in the biofiIn1s were then measured with microelectrodes prior to rapid freezing and slicing on a cryo-microtome. The vertical slices were stained with three different probes: a sulfate reducing bacterial (inclusive for most species of the 8-group of purple bacteria), a Desulfobacter, and a non-hybridising control probe. A good correlation between the vertical distribution of SRB and the measured 02-profile was found, with the SRB being restricted to the deeper anaerobic layers

Community structure of filamentous, sheath-building sulfur bacteria, Thioploca spp, off the coast of Chile

The filamentous sulfur bacteria Thioploca spp, produce dense bacterial mats in the shelf area off the coast of Chile and Peru. The mat consists of common sheaths, shared by many filaments, that reach 5 to 10 cm dean into the sediment, The structure of the Thioploca communities off the Bay of Concepcion was investigated,vith respect to biomass, species distribution, and three-dimensional orientation of the sheaths, Thioploca sheaths and filaments were found across the whole shelf area within the oxygen minimum zone, The maximum wet weight of sheaths, 800 g m(-2), was found at a depth of 90 m, The bacterial filaments within the sheaths contributed about 10% of this weight, The highest density of filaments was found within the uppermost I cm of the mat, On the basis of diameter classes, it was possible to distinguish populations containing only Thioploca spp, from mixed populations containing Beggiatoa spp, Three distinct size classes of Thioploca spp. were found, two of which have been described previously as Thioploca araucae and Thioploca chileae. Many Thioploca filaments did not possess a visible sheath, and about 20% of the sheaths contained more than one Thioploca species. The three-dimensional sheath structure showed that Thioploca filaments can move from the surface and deep into the sediment