Generation Of Analysis Tools Research Articles

High-Yield Expression in Escherichia coli, Purification and Application of Budding Yeast K2 Killer Protein

Saccharomyces cerevisiae K2 toxin is a highly active extracellular protein, important as a biocontrol agent for biotechnological applications in the wine industry. This protein is produced at negligible levels in yeast, making difficult to isolate it in amounts sufficient for investigation and generation of analysis tools. In this work, we demonstrate the use of a bacterial system for expression of the recombinant K2 protein, suitable for generation of antibodies specific for toxin of the yeast origin. Synthesis of the full-length S. cerevisiae K2 preprotoxin in Escherichia coli was found to be toxic to the host cell, resulting in diminished growth. Such effect was abolished by the introduction of the C-terminal truncation into K2 protein, directing it into non-toxic inclusion body fraction. The obtained protein is of limited solubility thus, facilitating the purification by simple and efficient chromatography-free procedure. The protein aggregates were successfully refolded into a soluble form yielding sufficient amounts of a tag-less truncated K2 protein suitable for polyclonal antibody production. Antibodies were raised in rabbit and found to be specific for detection of both antigen and native S. cerevisiae K2 toxin.

Metabolomics for the future of personalized medicine through information and communication technologies.

Although we have empirically found many ways to improve our lives, the challenge of cur­ ing diseases and extending our lifespan poses enormous problems for which we have, at the moment, no adequate answers. For instance, in spite of expenditures estimated to be on the order of €200 billion for cancer research over the last few decades, the definitive cure rates of most common forms of cancer have hardly increased. We will, therefore, need to take both the genetic and environmental effects differing between individuals much more into account. This will not only require a much more detailed characterization of each individual patient but will also require a new generation of ana lysis tools. We will need to convert much larger amounts of data into predictions on the effects of different therapies/prevention strategies. In turn, this will need to rely on a combination of mathematical and probabilistic modeling and advanced statistical inferential methodologies such as multivariate statistics and machine learn­ ing. Therefore, current empirical approaches will have to be replaced (or extended) by compu­ tational models adequate to take into account the complexity of the whole human being. A futuristic, data­intensive strategy to imple­ ment these ideas in the personalized medicine of the future would be the development of an individual model that, gathering together all the data available from the ’omic sciences, will build a ‘virtual patient’ associated to each individual. In this context, metabolomics will have a particular role with respect to the other ’omic sciences, because of its ability to detect, in real time, the adaptive multiparametric response of the organisms to pathophysiological stimuli or genetic modifications [1].

To Make Better Therapeutics, Companies Strive to Increase the Content of Results and Get them Faster.

To Make Better Therapeutics, Companies Strive to Increase the Content of Results and Get them Faster.

Research Needs in Lifeline Earthquake Engineering

Twenty seven research needs were presented at a workshop sponsored by ASCE, and are described and cross-referenced. The needs are referenced by type of research, area of impact, appropriate personnel, and appropriate sponsor. Topics include risk modelling, ground motion effects, equipment and facility response, and system analysis. Research types include analytical, experimental, data collection and product development. Areas of impact include the generation of analysis tools, data assembly, design procedure development, and planning methodologies. The aim is to acquaint researchers, research users, and research sponsors with a spectrum of research topics which are perceived to be of particular merit.