Project description:Arctic Mesorhizobium strain N33 was isolated from nodules of the Oxytropis arctobia in Canada’s eastern Arctic. This symbiotic bacterium can grow from 0 to 30°C, is one of the best known cold-adapted rhizobia, and can fix nitrogen at ~10°C. Here, the key molecular mechanisms of cold adaptation were investigated by determining changes in transcript profiles when cells were treated under eight different temperature conditions, including both sustained and transient cold treatments compared with cells grown at room temperature.
Project description:The complete genome of Polaribacter sp. NJDZ03, which was isolated from the surface of Antarctic macroalgae, was analyzed by next-generation sequencing, and a putative carrageenase gene Car3206 was obtained. Car3206 was cloned and expressed in Escherichia coli BL21(DE3). After purification by Ni-NTA chromatography, the recombinant Car3206 protein was characterized and the antioxidant activity of the degraded product was investigated. The results showed that the recombinant plasmid pet-30a-car3206 was highly efficiently expressed in E. coli BL21(DE3). The purified recombinant Car3206 showed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular weight of 45 kDa. The optimum temperature of the recombinant Car3206 was 55°C, and it maintain 60-94% of its initial activity for 4-12 h at 55°C. It also kept almost 70% of the initial activity at 30°C, and more than 40% of the initial activity at 10°C. These results show that recombinant Car3206 had good low temperature resistance and thermal stability properties. The optimum pH of recombinant Car3206 was 7.0. Car3206 was activated by Na+, K+, and Ca2+, but was significantly inhibited by Cu2+ and Cr2+. Thin-layer chromatographic analysis indicated that Car3206 degraded carrageenan generating disaccharides as the only products. The antioxidant capacity of the degraded disaccharides in vitro was investigated and the results showed that different concentrations of the disaccharides had similar scavenging effects as vitamin C on O2•- , •OH, and DPPH•. To our knowledge, this is the first report about details of the biochemical characteristics of a carrageenase isolated from an Antarctic Polaribacter strain. The unique characteristics of Car3206, including its low temperature resistance, thermal stability, and product unity, suggest that this enzyme may be an interesting candidate for industrial processes.
Project description:The bacterial genus Polaribacter is distributed widely in marine environments; however, there have been no reports of phages infecting Polaribacter strains. Here, we describe the isolation and genome sequencing of two lytic siphophages, P12002L and P12002S, that infect Polaribacter sp. strain IMCC12002. The two phages and host strain were isolated from coastal seawater of Korea. Complete genome sequences of the two phages were similar to each other and about 50 kb in length, with a G?+?C content of 28.9 %. The two genomes showed typical characteristics of phage genomes: a modular structure and high proportion of hypothetical proteins. The genome sequences have been deposited in GenBank under accession numbers KR136259 (P12002L) and KR136260 (P12002S).
Project description:C5aR1, a receptor for the complement activation proinflammatory fragment, C5a, is primarily expressed on cells of the myeloid lineage, and to a lesser extent on endothelial cells and neurons in brain. Previous work demonstrated C5aR1 antagonist, PMX205, decreased amyloid pathology and suppressed cognitive deficits in Alzheimer Disease (AD) mouse models. In the Arctic AD mouse model, genetic deletion of C5aR1 prevented behavior deficits at 10 months. However, the molecular mechanisms of this protection has not been definitively demonstrated. To understand the role of microglial C5aR1 in the Arctic AD mouse model, we have taken advantage of the CX3CR1GFP and CCR2RFP reporter mice to distinguish microglia as GFP-positive and infiltrating monocytes as GFP and RFP positive, for subsequent transcriptome analysis on specifically sorted myeloid populations from wild type and AD mouse models. Immunohistochemical analysis of mice aged to 2, 5, 7 and 10 months showed no change in amyloid beta (Ab) deposition in the Arctic C5aR1 knockout (KO) mice relative to that seen in the Arctic mice. Of importance, no CCR2+ monocytes/macrophages were found near the plaques in the Arctic brain with or without C5aR1. RNA-seq analysis on microglia from these mice identified inflammation related genes as differentially expressed, with increased expression in the Arctic mice relative to wildtype and decreased expression in the Arctic/C5aR1KO relative to Arctic. In addition, phagosomal-lysosomal proteins and protein degradation pathways that were increased in the Arctic mice were further increased in the Arctic/C5aR1KO mice. These data are consistent with a microglial polarization state with restricted induction of inflammatory genes and enhancement of clearance pathways.
Project description:Transcriptomic sequencing was performed to obtain the key functional genes involved in the adaptation of oxidative stress induced by hydrogen peroxide (H2O2) in the Arctic bacterium Pseudoalteromonas sp. A2. Exposure to 1 mmol/L H2O2 resulted in large alterations of the transcriptome profile, including significant upregulation of 109 genes and significant downregulation of 174 genes. Functional classification of differentially expressed genes revealed that most of genes affiliated with biological adhesion, negative regulation of biological process, enzyme regulator activity, protein binding transcription factor activity and structural molecular activity were upregulated, and most of genes affiliated with multicellular organismal process and extracellular region were downregulated. It was notably that fifteen genes affiliated with flagella and four genes affiliated with heat shock proteins were significantly upregulated. Meanwhile, nine genes affiliated with cytochrome and cytochrome oxidase, and five genes affiliated with TonB-dependent receptor, were significantly downregulated. However, eighteen genes with antioxidant activity categorized by GO analysis showed differential expressions. This overall survey of transcriptome and oxidative stress-relevant genes can contribute to understand the adaptive mechanism of Arctic bacteria. five significant upregulated genes and five significant downregulated genes were selected using qRT-PCR to cinduct the oxidative stress. overall survey of transcriptomic sequencing by RNA-Seq of the Pseudoalteromonas sp. A2, an isolate from seawater with high activity against H2O2
Project description:The objective of this study was to identify the different functional genes involved in key biogeochemical cycles in thehigh Arctic regions. Understanding the microbial diversity in the Arctic region is an important step to determine the effects of climate change on these areas.
Project description:To carry out population genetics analyses of the Arctic gregion we carried out Illumina Bead-Array-based enotyping on 18 samples from Greenland.