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Additional file 3: Figure S3. Western Blot membranes of ITM2A relative quantification expression in new born mice. Signal was detected by antibody anti-ITM2A AF4876. HEK293 ITM2A mouse GFP is used as control. On the left a-tubulin T9026 detection on the right anti-ITM2A AF4876. Detection with primary antibody was followed by HRP coupled with appropriate antibody. Then, luminescence was quantified.

Additional file 4: Figure S4. Detection limit of mouse ITM2A by western blot in HEK293 overexpressing mITM2A. Signal is quantified with MultiGauge v3.0. Housekeeping protein is ��-tubulin in orange and protein of interest is ITM2A in blue.

Additional file 2: Figure S2. Western Blot membrane of relative quantification of ITM2A protein expression in different cells type. Signal was detected by antibody anti-ITM2A AF4876 followed by HRP coupled anti-sheep antibody. Then, luminescence was quantified.

Additional file 1: Figure S1. Clonal selection of stable HEK293 overexpressing ITM2A. After fluorescence selection, better clones were confirmed by Western Blot. Clones with higher expression and higher luminescent signal were selected. Selected clones are circled in red. A HEK293 ITM2A human C-Ter and Nter GFP clone selection. B HEK293 ITM2A murin Nter GFP clone selection. C HEK293 ITM2A murin C-Ter GFP clone selection. D HEK293 ITM2A human C-Ter HA clone selection. E HEK293 ITM2A human N-Ter HA and WT clone selection.

The last two decades have seen a lot of research on context-aware and context-oriented software development technologies, across subfields of computer science. This research area is slowly starting to mature and researchers currently explore how to unify different solutions proposed in these subfields. We envision that within another decade some of these solutions will make it into mainstream software development approaches, tools and environments. Most end-user software built by that time will be context-aware and able to adapt seamlessly to its context of use (devices, surrounding environment, and users’ preferences). This transition from traditional to context-oriented software also requires a mindset shift in users. If users are to accept adaptive systems, they need to be in control. Context orientation should evolve to become less technology and more user-centric, putting the user back in control. A first step is to provide good feedback to the user about when and what adaptations take place, and mechanisms to allow users to partly control certain adaptations, followed by easily usable and understandable personalisation mechanisms dedicated to each end user. Eventually, when adaptive systems become completely natural and adopted by end users, this will culminate in our vision where users are in full control of relevant features or adaptations of applications of their interest, selected on-demand from online feature clouds, and integrated automatically into the running system.

Oomycetes in the class Saprolegniomycetidae of the Eukaryotic kingdom Stramenopila have evolved as severe pathogens of amphibians, crustaceans, fish and insects, resulting in major losses in aquaculture and damage to aquatic ecosystems. We have sequenced the 63 Mb genome of the fresh water fish pathogen, Saprolegnia parasitica. Approximately 1/3 of the assembled genome exhibits loss of heterozygosity, indicating an efficient mechanism for revealing new variation. Comparison of S. parasitica with plant pathogenic oomycetes suggests that during evolution the host cellular environment has driven distinct patterns of gene expansion and loss in the genomes of plant and animal pathogens. S. parasitica possesses one of the largest repertoires of proteases (270) among eukaryotes that are deployed in waves at different points during infection as determined from RNA-Seq data. In contrast, despite being capable of living saprotrophically, parasitism has led to loss of inorganic nitrogen and sulfur assimilation pathways, strikingly similar to losses in obligate plant pathogenic oomycetes and fungi. The large gene families that are hallmarks of plant pathogenic oomycetes such as Phytophthora appear to be lacking in S. parasitica, including those encoding RXLR effectors, Crinkler's, and Necrosis Inducing-Like Proteins (NLP). S. parasitica also has a very large kinome of 543 kinases, 10% of which is induced upon infection. Moreover, S. parasitica encodes several genes typical of animals or animal-pathogens and lacking from other oomycetes, including disintegrins and galactose-binding lectins, whose expression and evolutionary origins implicate horizontal gene transfer in the evolution of animal pathogenesis in S. parasitica.

Oomycetes in the class Saprolegniomycetidae of the Eukaryotic kingdom Stramenopila have evolved as severe pathogens of amphibians, crustaceans, fish and insects, resulting in major losses in aquaculture and damage to aquatic ecosystems. We have sequenced the 63 Mb genome of the fresh water fish pathogen, Saprolegnia parasitica. Approximately 1/3 of the assembled genome exhibits loss of heterozygosity, indicating an efficient mechanism for revealing new variation. Comparison of S. parasitica with plant pathogenic oomycetes suggests that during evolution the host cellular environment has driven distinct patterns of gene expansion and loss in the genomes of plant and animal pathogens. S. parasitica possesses one of the largest repertoires of proteases (270) among eukaryotes that are deployed in waves at different points during infection as determined from RNA-Seq data. In contrast, despite being capable of living saprotrophically, parasitism has led to loss of inorganic nitrogen and sulfur assimilation pathways, strikingly similar to losses in obligate plant pathogenic oomycetes and fungi. The large gene families that are hallmarks of plant pathogenic oomycetes such as Phytophthora appear to be lacking in S. parasitica, including those encoding RXLR effectors, Crinkler's, and Necrosis Inducing-Like Proteins (NLP). S. parasitica also has a very large kinome of 543 kinases, 10% of which is induced upon infection. Moreover, S. parasitica encodes several genes typical of animals or animal-pathogens and lacking from other oomycetes, including disintegrins and galactose-binding lectins, whose expression and evolutionary origins implicate horizontal gene transfer in the evolution of animal pathogenesis in S. parasitica.

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International audience

With the aim to obtain a cDNA coding for a mammalian methotrexate resistant dihydrofolate reductase (Dhfr) a plasmid ( pQS1 ) harboring the mouse wild type Dhfr cDNA was constructed and used to transform a methotrexate sensitive bacteria: B. subtilis. A plasmid, pQS4 , expressing large amount of Dhfr in both E. coli and B. subtilis was isolated through a two steps selection with two substrate analogues, trimethoprim followed by methotrexate. This new plasmid has a 54 bp duplication including the beta-lactamase promoter and a deletion of 564 bp removing the 5' end of the beta-lactamase coding region. These changes create a new -35 region TTGAAA and a potentially stronger binding site for both E. coli and B. subtilis 16S ribosomal RNA. pQS4 transformed B. subtilis were then grown in the presence of high level of methotrexate and resistant mutants isolated. One of them, pQS6 , which codes for an enzyme about 50 times more resistant to methotrexate than the wild type Dhfr was sequenced. It shows that a point mutation replaces the glutamine residue at position 35 by a proline.