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Search Results: 1 - 10 of 456 matches for " Ramona Gruetzner "
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Golden Gate Shuffling: A One-Pot DNA Shuffling Method Based on Type IIs Restriction Enzymes
Carola Engler, Ramona Gruetzner, Romy Kandzia, Sylvestre Marillonnet
PLOS ONE , 2009, DOI: 10.1371/journal.pone.0005553
Abstract: We have developed a protocol to assemble in one step and one tube at least nine separate DNA fragments together into an acceptor vector, with 90% of recombinant clones obtained containing the desired construct. This protocol is based on the use of type IIs restriction enzymes and is performed by simply subjecting a mix of 10 undigested input plasmids (nine insert plasmids and the acceptor vector) to a restriction-ligation and transforming the resulting mix in competent cells. The efficiency of this protocol allows generating libraries of recombinant genes by combining in one reaction several fragment sets prepared from different parental templates. As an example, we have applied this strategy for shuffling of trypsinogen from three parental templates (bovine cationic trypsinogen, bovine anionic trypsinogen and human cationic trypsinogen) each divided in 9 separate modules. We show that one round of shuffling using the 27 trypsinogen entry plasmids can easily produce the 19,683 different possible combinations in one single restriction-ligation and that expression screening of a subset of the library allows identification of variants that can lead to higher expression levels of trypsin activity. This protocol, that we call ‘Golden Gate shuffling’, is robust, simple and efficient, can be performed with templates that have no homology, and can be combined with other shuffling protocols in order to introduce any variation in any part of a given gene.
Assembly of Designer TAL Effectors by Golden Gate Cloning
Ernst Weber,Ramona Gruetzner,Stefan Werner,Carola Engler,Sylvestre Marillonnet
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0019722
Abstract: Generation of customized DNA binding domains targeting unique sequences in complex genomes is crucial for many biotechnological applications. The recently described DNA binding domain of the transcription activator-like effectors (TALEs) from Xanthomonas consists of a series of repeats arranged in tandem, each repeat binding a nucleotide of the target sequence. We present here a strategy for engineering of TALE proteins with novel DNA binding specificities based on the 17.5 repeat-containing AvrBs3 TALE as a scaffold. For each of the 17 full repeats, four module types were generated, each with a distinct base preference. Using this set of 68 repeat modules, recognition domains for any 17 nucleotide DNA target sequence of choice can be constructed by assembling selected modules in a defined linear order. Assembly is performed in two successive one-pot cloning steps using the Golden Gate cloning method that allows seamless fusion of multiple DNA fragments. Applying this strategy, we assembled designer TALEs with new target specificities and tested their function in vivo.
A Modular Cloning System for Standardized Assembly of Multigene Constructs
Ernst Weber,Carola Engler,Ramona Gruetzner,Stefan Werner,Sylvestre Marillonnet
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0016765
Abstract: The field of synthetic biology promises to revolutionize biotechnology through the design of organisms with novel phenotypes useful for medicine, agriculture and industry. However, a limiting factor is the ability of current methods to assemble complex DNA molecules encoding multiple genetic elements in various predefined arrangements. We present here a hierarchical modular cloning system that allows the creation at will and with high efficiency of any eukaryotic multigene construct, starting from libraries of defined and validated basic modules containing regulatory and coding sequences. This system is based on the ability of type IIS restriction enzymes to assemble multiple DNA fragments in a defined linear order. We constructed a 33 kb DNA molecule containing 11 transcription units made from 44 individual basic modules in only three successive cloning steps. This modular cloning (MoClo) system can be readily automated and will be extremely useful for applications such as gene stacking and metabolic engineering.
Verdade e reconcilia??o para as " gera??es roubadas": revisitando a história da Austrália
Vijeyarasa, Ramona;
Sur. Revista Internacional de Direitos Humanos , 2007, DOI: 10.1590/S1806-64452007000200006
Abstract: the tasmanian state government and the australian federal senate have taken recent steps towards setting up a reparations tribunal for aboriginal and torres strait islander (atsi) people who were separated from their families and communities under state-based forced removal policies of the 20th century. this paper proposes a truth and reconciliation commission drawing on international lessons.
Formal Institutions and Regional Development. Considerations Regarding Romania
Ramona FRUNZ?
Theoretical and Applied Economics , 2011,
Abstract: Assuming that a theory of development is incomplete without a theory of institutions, we propose in this paper to analyze, in a first part, the interrelation formal institutions-regional development and the ways trough which these two components sustain themselves, then, in the second part of the paper, we will analyze the degree of convergence of the eight development regions in Romania, taking into account indicators such as: GDP per capita, average net wage gain, unemployment rate, labor productivity, the share of sectors in total employment.The results of our research we hope to lead us, in the final of the paper, to draw some future action directions by which to reduce development disparities between regions, both at national level and at European level NUTS2.
The XX century architectural heritage in the Marche region. The Masters’ drawings and designs. Il patrimonio dell’architettura del XX secolo nelle Marche. Disegni e progetti dei Maestri .
Ramona Quattrini
Abstract: In the history of architecture, Marche is often considered as a region that did not take part in the big developments of national architecture. Hence it’s meaningful to examine the architectonic events of the Region in the XX century and how much the Marche region has been the object of work by the great masters of the past century. A list of architects - called ‘masters’ – has been selected: Franco Albini, Gae Aulenti, Carlo Aymonino, Ernesto Basile, BBPRGuido Canella, Giancarlo De Carlo, Luigi Figini, Massimiliano Fuksas, Roberto Gabetti, Ignazio Gardella, Giorgio Grassi, Vittorio Gregotti, Adalberto Libera, Giovanni Michelucci, Gaetano Minnucci, Carlo Mollino, Luigi Moretti, Pier Luigi Nervi, Giuseppe Pagano, Edoardo Persico, Marcello Piacentini, Renzo Piano, Gio Ponti, Paolo Portoghesi, Franco Purini, Ludovico Quaroni, Mario Ridolfi, Aldo Rossi, Giuseppe Samonà, Antonio Sant’Elia; Alberto Sartoris, Carlo Scarpa, Giuseppe Terragni, Gino Valle, Vittoriano Viganò, Marco Zanuso, The research is a meaningful census of the works of the 20th century, whether they have been realized or not (projects, competitions, studies etc.) and assigned to the masters listed above. Il presente articolo raccoglie e documenta in parte gli esiti della tesi di dottorato della scrivente, condotta con la guida del Prof. Paolo Clini. Nella considerazione di una realtà di provincia, la tesi si è prefissata di indagare gli esiti architettonici del XX secolo nelle Marche a partire da una rosa di architetti, definiti con il termine maestri[i]. La ricerca si proponeva un censimento esaustivo e significativo delle loro opere, realizzate e non (progetti, concorsi, studi, etc…). [i] Vengono qui riportati i nominativi dei progettisti inclusi nell’elenco: Albini, Aulenti, Aymonino, Basile, BBPR, Canella, De Carlo, Figini, Fuksas, Gabetti, Gardella, Grassi, Gregotti, Libera, Michelucci, Minnucci, Mollino, Moretti, Nervi, Pagano, Persico, Piacentini, Piano, Ponti, Portoghesi, Purini, Quaroni, Ridolfi, Rossi, Samonà, Sant’elia; Sartoris, Scarpa, Terragni, Valle, Viganò, Zanuso.
Ilse Lenz (Hg.): Die Neue Frauenbewegung in Deutschland. Wiesbaden: VS Verlag für Sozialwissenschaften 2008
Ramona Bechler
querelles-net , 2009,
Abstract: Mit über 250 zusammengetragenen Quellen versammelt und dokumentiert die Herausgeberin Ilse Lenz eine Vielzahl von Str mungen und Themenfeldern aus 40 Jahren deutscher Frauenbewegung. Eingebettet in fundierte Einführungstexte bietet jedes Kapitel des Bandes bekannte und weniger bekannte Ver ffentlichungen von Akteur/-innen der Frauenbewegung. Als innovativ kann die Einbeziehung der u erungen emanzipierter M nner in die Betrachtung gewertet werden. Das weitgehende Fehlen der staatsunabh ngigen Frauenbewegung in der DDR ist dagegen ein bedauerlicher Mangel. Dennoch darf die vorliegende Quellensammlung als zukünftiges Standardwerk für die Geschichte der deutschen Frauenbewegung gelten. The editor Ilse Lenz collects documents gathered together from multiple thematic trends and fields representing 40 years of the women’s movement in Germany. Embedded in ground-laying introductory texts, each chapter of the volume offers well-known and less known publications by those who took part in the women’s movement. The decision to include emancipated men in the examination can be seen as innovative while the largely absent autonomous women’s movement in the GDR can be seen as an unfortunate flaw. Nevertheless, the source book at hand can be considered a standard work for the history of the German women’s movement.
Subsidies in Croatian agriculture and adjustment to the EU
Ramona Frani?
Zbornik Radova Ekonomskog Fakulteta u Rijeci : ?asopis za Ekonomsku Teoriju i Praksu , 2005,
Abstract: After the Croatian independence and re-adoption of the free trade principles, in political debates agriculture got the role of one of the basic pillar in economic development. There is intention to protect domestic agriculture against much more developed western European agriculture and to support its competitiveness, respecting at the same time the requirements of the international trade agreements asking for decreasing domestic protection. The reform of agricultural price policy started in 1988, due to the trade policy reform within the requirements imposed by the membership in the World Trade Organization. Recent changes from 2003 tend to simplify the system of agricultural support and to equalize the level of incentives. Together with existing production subsidies, new models of income support, capital investments and rural development are introduced. The reform predicts harmonization of domestic agricultural policy measures with those conducted or should be conducted in the EU. It means the shift from the old way of supporting European agricultural markets, toward incentives for European farmer, in the way that in total support prevails income, and not production support.
Tesu Ramona
Annals of the University of Oradea : Economic Science , 2012,
Abstract: The present article has as a reserch field theoretical, methodological and practical aspects of the own funds, taking into account the line-up attempts of the romanian banking system to the requirements of the European Union, materialised in the Basel Agreement. The objectives of the research theme, presented in this paper, are mainly oriented towards own funds. We also proposed the following secondary objectives: presenting the concept approaches concerning own funds, emphasysing the tipology of own funds and pointing out their purpose in the banking activity; presenting and analyse the evolution of the own funds in Romania, for a period of 8 years (2004-2011). Even though the actual stage of the research in this field is advanced, in the romanian and foreign literature which dedicate a lot of theoretical and empiric studies concerning own funds. In our study we will use a theoretical and a practical research. The theoretical research describes the laws, reglementations and rules of application in the banking field in our country. In the practical part we will use as a research method the quality-comparative analyse by presenting details regarding bank back-ups. Despite the global financial crisis, the Romanian banking system can be considered as stable as a whole, with levels of capitalization, solvency and liquidity in accordance with consistent with prudential requirements. Tier 1, defined as the core of their sources of credit institutions through the permanent criteria, payment flexibility and capacity to absorb losses continued to overwhelmingly support a total equity. Appropriate quality of own funds is on their structure. The most important component frame our capital plus capital frame attached. The second component in terms is still represented by legal reserves. An important challenge for banks was to maintain a level of own funds accordance, in the conditions witch they were eroded by the current period losses. Tier 2 support rate of about 20 percentages of total equity and subordinated debt were the main components and revaluation reserves. Re-evaluation reserves have preserved funding contributions to Tier 2. An for evaluation of the level of capitalization of credit institutions, the central bank uses solvable indicator, as the only indicator currently regulated for this purpose. It also provides banks in Romania at the conditions for additional capital requirements related to Basel III framework to be taken by the European regulatory Capital Requirement Directive IV. As a final conclusion of the research in the own funds field we want t
PDF/A standard for long term archiving
Ramona Vasilescu
Annals. Computer Science Series , 2009,
Abstract: PDF/A is defined by ISO 19005-1 as a file format based on PDF format. The standard provides a mechanism for representing electronic documents in a way that preserves their visual appearance over time, independent of the tools and systems used for creating or storing the files.
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