11 research outputs found
Systems Approach to Arms Control Verification
Using the decades of experience of developing concepts and technologies for verifying bilateral and multilateral arms control agreements, a broad conceptual systems approach is being developed that takes into account varying levels of information and risk. The IAEA has already demonstrated the applicability of a systems approach by implementing safeguards at the state level, with acquisition path analysis as the key element. In order to test whether such an approach could also be implemented for arms control verification, an exercise was conducted in November 2014 at the European Commission Joint Research Centre, Ispra, Italy. Based on the scenario of a hypothetical treaty between two model nuclear weapons states that aims to cap their nuclear arsenals at existing levels, the goal of this exercise was to explore how to use acquisition path analysis (APA) in an arms control context. Our contribution will present the scenario, objectives and results of this exercise
Developing a Systems Concept to Arms Control Verification — Results from the ESARDA VTM WG Workshop
Applying State-level Approaches to Arms Control Verification – a Systems Concept, VTM Working Group Workshop Summary
Enhanced plasmid production in miniaturized high-cell-density cultures of Escherichia coli supported with perfluorinated oxygen carrier
Optimization of fermentation conditions for an Escherichia coli strain engineered using the response surface method to produce a novel therapeutic DNA vaccine for rheumatoid arthritis
De novo creation of MG1655-derived E. coli strains specifically designed for plasmid DNA production
The interest in plasmid DNA (pDNA) as a biopharmaceutical has been increasing over the last several years, especially after the approval of the first DNA vaccines. New pDNA production strains have been created by rationally mutating genes selected on the basis of Escherichia coli central metabolism and plasmid properties. Nevertheless, the highly mutagenized genetic background of the strains used makes it difficult to ascertain the exact impact of those mutations. To explore the effect of strain genetic background, we investigated single and double knockouts of two genes, pykF and pykA, which were known to enhance pDNA synthesis in two different E. coli strains: MG1655 (wild-type genetic background) and DH5α (highly mutagenized genetic background). The knockouts were only effective in the wild-type strain MG1655, demonstrating the relevance of strain genetic background and the importance of designing new strains specifically for pDNA production. Based on the obtained results, we created a new pDNA production strain starting from MG1655 by knocking out the pgi gene in order to redirect carbon flux to the pentose phosphate pathway, enhance nucleotide synthesis, and, consequently, increase pDNA production. GALG20 (MG1655ΔendAΔrecAΔpgi) produced 25-fold more pDNA (19.1 mg/g dry cell weight, DCW) than its parental strain, MG1655ΔendAΔrecA (0.8 mg/g DCW), in glucose. For the first time, pgi was identified as an important target for constructing a high-yielding pDNA production strain.MIT-Portugal ProgramFundação para a Ciência e a Tecnologia (project PTDC/ EBB-EBI/113650/2009, PhD grant SFRH/BD/33786/2009