Boreal peatlands with low iron availability are a potential, but rarely studied, source for the isolation of bacteria for applications in metal sorption. The present research focused on the isolation and identification of Actinobacteria from northern Finland, which can produce siderophores for metal capture. The 16S rDNA analysis showed that isolated strains belonged to Firmicutes (Bacillus sp.) and Actinobacteria (Microbacterium sp.). The culture most efficiently producing siderophores in the widest array of the media was identified as Microbacterium sp. The most appropriate media for siderophore production by the Microbacterium strain were those prepared with glucose supplemented with asparagine or glutamic acid, and those prepared with glycerol or fructose supplemented with glutamic acid. The microorganism obtained and its siderophores were used to develop Sphagnum moss-based hybrid biosorbents. It was showed that the hybrid sorbent could bind nickel ions and that the nickel removal was enhanced by the presence of siderophores. Bacterial cells did not have a significant effect on sorption efficiency compared to the use of siderophores alone. The microbial biosorbent could be applied in the final effluent treatment stage for wastewater with low metal concentrations.
The polymerase chain reaction (PCR), one of the most commonly applied methods of diagnostics and molecular biology has a frustrating downside known as the false positive signal or contamination. Several solutions to avoid and to eliminate PCR contaminations have been worked out to date but the implementation of these solutions to laboratory practice may be laborious and time consuming. A simple approach to circumvent the problem of persisting PCR contamination is reported. The principle of this approach lies in shortening the steps of denaturation, annealing, and elongation in the PCR thermal cycle. The modification leads to the radical decline of false positive signals obtained for the no-template controls without affecting the detection of target PCR products. In the model experiments presented here, the signal of negative control was shifted by about ten cycles up above those for the examined samples so that it could be neglected. We do not recommend this solution in PCR diagnostics, where the sensitivity of detection is of the highest priority. However, the approach could be useful to pass by the problem of persisting contamination in quantitative PCR, where the range of quantitation is usually much above the limits of detection.Electronic supplementary materialThe online version of this article (doi:10.1007/s13353-015-0336-z) contains supplementary material, which is available to authorized users.
A simple microplate method was designed for rapid testing DNA-binding activity of proteins. The principle of the assay involves binding of tested DNA by his-tagged protein immobilized on a nickel-coated ELISA plate, following colorimetric detection of biotinylated DNA with avidin conjugated to horseradish peroxidase. The method was used to compare DNA mismatch binding activities of MutS proteins from three bacterial species. The assay required relatively low amounts of tested protein (approximately 0.5–10 pmol) and DNA (0.1–10 pmol) and a relatively short time of analysis (up to 60 min). The method is very simple to apply and convenient to test different buffer conditions of DNA–protein binding. Sensitive colorimetric detection enables naked eye observations and quantitation with an ELISA reader. The performance of the assay, which we believe is a distinguishing trait of the method, is based on two strong and specific molecular interactions: binding of a his-tagged protein to a nickel-coated microplate and binding of biotinylated DNA to avidin. In the reported experiments, the solution was used to optimize the conditions for DNA mismatch binding by MutS protein; however, the approach could be implemented to test nucleic acids interactions with any protein of interest.
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