Draft Genome Sequence of Streptomyces sp. Strain ventii, Isolated from a Microbial Mat near Hydrothermal Vents within the Axial Seamount in the Pacific Ocean, and Resequencing of the Type Strains Streptomyces lonarensis NCL 716 and Streptomyces bohaiensis 11A07

The draft genome of Streptomyces sp. strain ventii, an environmental isolate recovered from deep-sea hydrothermal vents in the Pacific Ocean, is presented along with the resequenced draft genomes of the type strains Streptomyces bohaiensis 11A07 and Streptomyces lonarensis NCL 716

Characterization of Lipid-Anchored Inhibitor Rulers as a Measure of Enzyme Topography in Coagulation Enzyme, Factor X

Background: The blood coagulation cascade (BCC) is activated under various circumstances. An injury is a good example of how the process begins. This system is a tightly regulated series of events. The enzymes involved assemble with their respective cofactors on lipid membranes to reach their full pro-coagulant complex potential. Each of these factors is part of the intrinsic pathway and both are required for the activation of factor X. Factor Xa is a very important part of the blood coagulation cascade because its activation is primarily responsible for thrombin generation. Factor Xa, anchored in the plasma membrane, forms a complex with other proteins for example, Factor V. The topography of the Factor Xa alone, and in complex with other factors on the cell surface is poorly understood. This study will focus particularly on the activated form of factor X (fXa).This study will focus particularly on the activated form of factor X (fXa). FX is of the utmost importance in coagulation because it is integral in the generation of thrombin from prothrombin. Kunitz-type protein inhibitors (KPI) are globulin proteins which inhibit trypsin much like the ones that regulate FX. Basic pancreatic trypsin inhibitor (BPTI) is the kunitz-protein inhibitor of particular interest in this study. The goal is to gain understanding of the topography of fXa in the presence of BPTI. More specifically, we will examine the range of reactive heights of the blood clotting factor X/BPTI complex above the membrane surface. Methods: Previous studies suggest that the height and topology of the active site of FXa above the plasma membrane is of importance to its function. Thus, we would like to generate a series of lipid-anchored ‘inhibitor rulers’ to measure the height of the FXa complex above the plasma membrane. A construct encoding BPTI fused with a bacterial lipid-anchor consensus site (LAGC) separated by ten linker sequences (a total of 75Å) is being built. More specifically, a segment coding for an LAGC leader sequence and EA3K linker sequences is being directionally cloned into pET11d E. coli DNA. Oligonucleotides were designed encoding the EA3K linker repeat and are being introduced into the constructed pET11d plasmid via the engineered restriction endonuclease sites existing between the LAGC sequence and the first part of the BPTI sequence. Next, these lipid-anchored inhibitor rulers will be cloned, expressed, and purified using a series of affinity chromatography steps. Specifically, we will immobilize the enzyme trypsin to the solid support, and use this to separate and purify our newly generated inhibitor product. Once the ruler has been constructed, we will use it to determine the height above the cell membrane for FXa, individually and in the context of their pro-coagulant complexes. Using an equilibrium-based chromogenic enzyme assay, we will measure the rates and extent of inhibition for both FXa and the FXa/pro-coagulant complex. The long-term goal will be to change the length of the ruler and measure the effect on inhibition. Conclusions: These sequences have been introduced to the plasmid DNA and ligation experiments using a T4 DNA ligase at a 3:1 insert to vector ratio are taking place. Upon ligation, this DNA will be transformed into E. coli DH5α competent cells. Preliminary tests are being done to confirm any colonies with the proper sequence. If any colonies are confirmed preliminarily, they will be sent for DNA sequencing. Upon sequencing confirmation, we will transform the plasmid into an expression strain and begin phase 2 of the project

Energy and Climate Change: Recommendations for the City of Springfield Regarding Buildings, Electricity, and Transportation

36 pagesThe City of Springfield is interested in learning about its current environmental impacts and ways that it might improve its environmental footprint in the future. The students of University of Oregon course PPPM 607: Energy and Climate Change researched three topics—buildings, electricity, and transportation— related to the City of Springfield’s influence on energy use, climate emissions, and quality of life of the community. Based on this research, groups of students made recommendations in these three areas for how the City of Springfield could reduce environmental impacts

Electric Vehicle design for Shell ECO-Marathon

With recent focus on transportation energy and CO2 emissions, an interest has emerged in finding innovative methods to reduce greenhouse gas emissions from petroleum based fuels. Many alternative fuel options are being researched, with one being electric vehicles (EVs). EVs offer many advantages over typical petroleum-based forms of transportation. They produce no emissions and have high efficiency. Additionally EVs can be charged using alternative power sources, such as wind and solar, making them a truly renewable source of transportation. For 3 year, the Boise State University Horsepower team has been designing, constructing, and competing an electric vehicle in the Shell ECO-Marathon in Houston, Texas. This competition brings together teams from all over the North and South America to test their alternative energy vehicle designs. The winner of the 2012 competition achieved a gas mileage equivalent of 2,189 mpg, almost enough to drive from Boise, Idaho to Houston, Texas on one gallon of gasoline. This year’s Boise State University Horsepower team is focused on improving the frame and shell of the vehicle, improving steering and braking capabilities, and integrating regeneration systems such as solar panels and regenerative braking. The new shell will significantly improve vehicle aerodynamics and reduce drag forces. The frame is being redesigned to fit inside the new shell and reduce overall weight. Improving the steering will reduce the energy wasted during turns due to a skidding problem that hampered the vehicle last year. In addition, regeneration systems will gather energy while the vehicle is operating, to reduce power consumption and boost efficiency. Through this process, the Boise State University Horsepower team aims to improve the efficiency of the current electric vehicle and place well at the competition