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Kópavogur, Iceland

Erlendsson L.S.,ORF genetics | Muench M.O.,University of California at San Francisco | Hellman U.,Uppsala University | Hrafnkelsdottir S.M.,ORF genetics | And 5 more authors.
Biotechnology Journal

Biologically active recombinant human Flt3 ligand was expressed and isolated from transgenic barley seeds. Its expression is controlled by a tissue specific promoter that confines accumulation of the recombinant protein to the endosperm tissue of the seed. The recombinant Flt3 ligand variant expressed in the seeds contains an HQ-tag for affinity purification on immobilized metal ion affinity chromatography (IMAC) resin. The tagged protein was purified from seed extracts to near homogeneity using sequential chromatography on IMAC affinity resin and cation exchange resin. We also show that the recombinant Flt3 ligand protein undergoes posttranslational modifications: it is a glycoprotein containing α-1,3-fucose and α-1,2-xylose. The HQ-tagged Flt3 ligand variant exhibits comparable biological activity to commercial Flt3 ligand. This is the first report showing expression and accumulation of recombinant human growth factor in barley seeds with a yield of active protein similar to a bacterial expression system. The present results demonstrate that plant molecular farming is a viable approach for the bioproduction of human-derived growth factors. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA. Source

Olafsson K.,University of Iceland | Olafsson K.,Matis Ltd. | Einarsson S.M.,Institute of Freshwater Fisheries | Gilbey J.,Freshwater Fisheries Laboratory | And 5 more authors.
ICES Journal of Marine Science

The origin and life history of 186 Atlantic salmon caught at sea within Icelandic waters were investigated using microsatellites to assess the origin and scales and otoliths to assess freshwater and sea age. A total of 184 samples were aged using scales or otolithes or both. Most of the samples were from individuals in their first year at sea (72.8%). The freshwater age varied from 1 to 5 years with an average of 2.6 years. The most common freshwater age was 2 years (42%), with a further substantial proportion of 3-year-old fish (28%). Genetic assignment of individual fish to their most likely population of origin was performed using Bayesian genetic individual assignments with a baseline consisting of 284 Eastern Atlantic rivers and 466 sample sites genotyped at the 14 microsatellite. A total of 186 samples of salmon caught at sea were assigned to their origin. Eight samples, from post-smolts and caught close to land, were assigned as having come from Iceland. Of the remaining 178 samples, 121 individuals (68%) were from the Southern Group, i.e. from mainland Europe, the UK, and Ireland, 53 individuals (30%) were from the Northern Group, i.e. Scandinavia and Northern Russia, and 4 individuals were from Iceland (2%). Stock mixture proportions were estimated for four periods using ONCOR and cBAYES. Stock mixture analysis generally supported the individual assignments, but did not suggest a seasonal component to the distribution of salmon stocks. These results indicate that the sea south and east of Iceland are important as feeding areas for migrating Atlantic salmon, particularly for salmon originating in the UK, Ireland, and southern Europe. Furthermore, the lack of adult Icelandic fish so close to Iceland is remarkable and suggests that Atlantic salmon from Icelandic stocks are using different feeding grounds. © 2015 International Council for the Exploration of the Sea 2015. All rights reserved. Source

Plotka M.,University of Gdansk | Kaczorowska A.-K.,University of Gdansk | Morzywolek A.,University of Gdansk | Makowska J.,University of Gdansk | And 12 more authors.

Phage vB-Tsc2631 infects the extremophilic bacterium Thermus scotoductus MAT2631 and uses the Ts2631 endolysin for the release of its progeny. The Ts2631 endolysin is the first endolysin from thermophilic bacteriophage with an experimentally validated catalytic site. In silico analysis and computational modelling of the Ts2631 endolysin structure revealed a conserved Zn2+ binding site (His30, Tyr58, His131 and Cys139) similar to Zn2+ binding site of eukaryotic peptidoglycan recognition proteins (PGRPs). We have shown that the Ts2631 endolysin lytic activity is dependent on divalent metal ions (Zn2+ and Ca2+). The Ts2631 endolysin substitution variants H30N, Y58F, H131N and C139S dramatically lost their antimicrobial activity, providing evidence for the role of the aforementioned residues in the lytic activity of the enzyme. The enzyme has proven to be not only thermoresistant, retaining 64.8% of its initial activity after 2 h at 95°C, but also highly thermodynamically stable (Tm = 99.82°C, ΔHcal = 4.58 × 104 cal mol-1). Substitutions of histidine residues (H30N and H131N) and a cysteine residue (C139S) resulted in variants aggregating at temperatures ≥75°C, indicating a significant role of these residues in enzyme thermostability. The substrate spectrum of the Ts2631 endolysin included extremophiles of the genus Thermus but also Gram-negative mesophiles, such as Escherichia coli, Salmonella Panama, Pseudomonas fluorescens and Serratia marcescens. The broad substrate spectrum and high thermostability of this endolysin makes it a good candidate for use as an antimicrobial agent to combat Gram-negative pathogens. Copyright © 2015 Plotka et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Source

Magnusdottir A.,ORF genetics | Vidarsson H.,ORF genetics | Bjornsson J.M.,ORF genetics | Orvar B.L.,ORF genetics
Trends in Biotechnology

The most popular hosts for recombinant protein production have now in many cases passed their low-hanging-fruit era and their limitations as production hosts are becoming more evident. Therefore, the bioprocessing community is constantly on the lookout for new hosts that can complement the current selection. The development of plant systems is eagerly followed because of the great potential they show, such as much reduced production cost and absence of endotoxins and human pathogens. In this review, we describe the unique barley-based platform for recombinant protein production. We summarize the methods used for gene modification and protein processing and discuss the main advantages of the system. Finally, we discuss the significance of endotoxin-free proteins for cell-based research and therapeutic applications. © 2013 Elsevier Ltd. Source

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