Construction of a Z-DNA-specific restriction endonuclease

Novel restriction enzymes can be created by fusing the nuclease domain of FokI endonuclease with defined DNA binding domains. Recently, we have characterized a domain (Zα) from the N-terminal region of human double-stranded RNA adenosine deaminase (hADAR1), which binds the Z-conformation with high specificity. Here we report creation of a conformation-specific endonuclease, Zα nuclease, which is a chimera of Zα and FokI nuclease. Purified Zα nuclease cleaves negatively supercoiled plasmids only when they contain a Z-DNA forming insert, such as (dC-dG)13. The precise location of the cleavage sites was determined by primer extension. Cutting has been mapped to the edge of the B-Z junction, suggesting that Zα nuclease binds within the Z-DNA insert, but cleaves in the nearby B-DNA, by using a mechanism similar to type IIs restriction enzymes. These data show that Zα binds Z-DNA in an environment similar to that in a cell. Zα nuclease, a structure-specific restriction enzyme, may be a useful tool for further study of the biological role of Z-DNA.

Construction of neocentromere-based human minichromosomes by telomere-associated chromosomal truncation

Neocentromeres (NCs) are fully functional centromeres that arise ectopically in noncentromeric regions lacking α-satellite DNA. Using telomere-associated chromosome truncation, we have produced a series of minichromosomes (MiCs) from a mardel(10) marker chromosome containing a previously characterized human NC. These MiCs range in size from ≈0.7 to 1.8 Mb and contain single-copy intact genomic DNA from the 10q25 region. Two of these NC-based Mi-Cs (NC-MiCs) appear circular whereas one is linear. All demonstrate stability in both structure and mitotic transmission in the absence of drug selection. Presence of a functional NC is shown by binding a host of key centromere-associated proteins. These NC-MiCs provide direct evidence for mitotic segregation function of the NC DNA and represent examples of stable mammalian MiCs lacking centromeric repeats.

Construction and characterization of an azurin analog for the purple copper site in cytochrome c oxidase.

A protein analog of a purple copper center has been constructed from a recombinant blue copper protein (Pseudomonas aeruginosa azurin) by replacing the loop containing the three ligands to the blue copper center with the corresponding loop of the CuA center in cytochrome c oxidase (COX) from Paracoccus denitrificans. The electronic absorption in the UV and visible region (UV-vis) and electron paramagnetic resonance (EPR) spectra of this analog are remarkably similar to those of the native CuA center in COX from Paracoccus denitrificans. The above spectra can be obtained upon addition of a mixture of Cu2+ and Cu+. Addition of Cu2+ only results in a UV-vis spectrum consisting of absorptions from both a purple copper center and a blue copper center. This spectrum can be converted to the spectrum of a pure purple copper by a prolonged incubation in the air, or by addition of excess ascorbate. The azurin mutant reported here is an example of an engineered purple copper center with the A480/A530 ratio greater than 1 and with no detectable hyperfines, similar to those of the CuA sites in COX of bovine heart and of Paracoccus denitrificans.

Spatiotemporal characteristics of the Huangtupo landslide in the Three Gorges region (China) constrained by radar interferometry

The Huangtupo landslide is one of the largest in the Three Gorges region, China. The county-seat town of Badong, located on the south shore between the Xiling and Wu gorges of the Yangtze River, was moved to this unstable slope prior to the construction of the Three Gorges Project, since the new Three Gorges reservoir completely submerged the location of the old city. The instability of the slope is affecting the new town by causing residential safety problems. The Huangtupo landslide provides scientists an opportunity to understand landslide response to fluctuating river water level and heavy rainfall episodes, which is essential to decide upon appropriate remediation measures. Interferometric Synthetic Aperture Radar (InSAR) techniques provide a very useful tool for the study of superficial and spatially variable displacement phenomena. In this paper, three sets of radar data have been processed to investigate the Huangtupo landslide. Results show that maximum displacements are affecting the northwest zone of the slope corresponding to Riverside slumping mass I#. The other main landslide bodies (i.e. Riverside slumping mass II#, Substation landslide and Garden Spot landslide) exhibit a stable behaviour in agreement with in situ data, although some active areas have been recognized in the foot of the Substation landslide and Garden Spot landslide. InSAR has allowed us to study the kinematic behaviour of the landslide and to identify its active boundaries. Furthermore, the analysis of the InSAR displacement time-series has helped recognize the different displacement patterns on the slope and their relationships with various triggering factors. For those persistent scatterers, which exhibit long-term displacements, they can be decomposed into a creep model (controlled by geological conditions) and a superimposed recoverable term (dependent on external factors), which appears closely correlated with reservoir water level changes close to the river's edge. These results, combined with in situ data, provide a comprehensive analysis of the Huangtupo landslide, which is essential for its management.

Archeological investigations along the proposed Alibates tour road improvement construction route, Alibates Flint Quarries National Monument, Potter County, Texas /

"Contract no. PX 7029-8-0986."

Final environmental impact statement : proposed issuance of Federal permits to the Pittston Company of New York for the construction of a 250,000 barrel/day oil refinery and marine terminal, Eastport, Maine /

Cover title.

Windstorm mitigation manual for light frame construction /

Sponsored by Federal Emergency Management Agency, Illinois Emergency Management Agency and State Farm Fire and Casualty Company.

Construction equipment ownership and operating expense schedule.

"September 1997."

Construction equipment ownership and operating expense schedule.

"August 1993."

Construction equipment ownership and operating expense schedule.

"August 1995."

Problematising the construction of journal quality:an engagement with the mainstream

Journal ranking studies have generally adopted citation techniques or academic perceptions as the basis for assessing journal quality. They have traditionally been a source of information about potential research outlets, new journals, and an aid to developing a consensus about the relative merit of publications for promotion decisions. The aim of our research is to address specific shortcomings in the conventional literature and construct an alternative view of how we might more appropriately assess journal ‘quality’. We attempt to engage with the conventional literature by applying an approach that does not privilege either citation techniques or academic perceptions. We have adopted from Zeff (1996) an objective measure of academic journal library holdings, which Zeff describes as a ‘market test’. Our construct provides evidence of an important difference in journal holdings for the Australasian region that could significantly influence further research on journal quality. The method itself is entirely mundane but may be considered to reflect a complex of historic and more contemporary variables which impact on academic and administrative decisions, influencing the makeup of academic library holdings and providing a proxy for journal ‘quality’.

Depth propagation and surface construction in 3-D vision

In stereo vision, regions with ambiguous or unspecified disparity can acquire perceived depth from unambiguous regions. This has been called stereo capture, depth interpolation or surface completion. We studied some striking induced depth effects suggesting that depth interpolation and surface completion are distinct stages of visual processing. An inducing texture (2-D Gaussian noise) had sinusoidal modulation of disparity, creating a smooth horizontal corrugation. The central region of this surface was replaced by various test patterns whose perceived corrugation was measured. When the test image was horizontal 1-D noise, shown to one eye or to both eyes without disparity, it appeared corrugated in much the same way as the disparity-modulated (DM) flanking regions. But when the test image was 2-D noise, or vertical 1-D noise, little or no depth was induced. This suggests that horizontal orientation was a key factor. For a horizontal sine-wave luminance grating, strong depth was induced, but for a square-wave grating, depth was induced only when its edges were aligned with the peaks and troughs of the DM flanking surface. These and related results suggest that disparity (or local depth) propagates along horizontal 1-D features, and then a 3-D surface is constructed from the depth samples acquired. The shape of the constructed surface can be different from the inducer, and so surface construction appears to operate on the results of a more local depth propagation process.

Making the construction industry resilient to extreme weather:lessons from construction in hot weather conditions

The construction industry is susceptible to extreme weather events (EWEs) due to most of its activities being conducted by manual workers outdoors. Although research has been conducted on the effects of EWEs, such as flooding and snowfall, limited research has been conducted on the effects of heatwaves and hot weather conditions. Heatwaves present a somewhat different risk profile to construction, unlike EWEs such as flooding and heavy snowfall that present physical obstacles to work onsite. However, heatwaves have affected the construction industry in the UK, and construction claims have been made due to adverse weather conditions. With heatwaves being expected to occur more frequently in the coming years, the construction industry may suffer unlike any other industry during the summer months. This creates the need to investigate methods that would allow construction activities to progress during hot summer months with minimal effect on construction projects. Hence, the purpose of this paper. Regions such as the Middle East and the UAE in particular flourish with mega projects, although temperatures soar to above 40̊C in the summer months. Lessons could be learnt from such countries and adapted in the UK. Interviews have been conducted with a lead representative of a client, a consultant and a contractor, all of which currently operate on UAE projects. The key findings include one of the preliminary steps taken by international construction companies operating in the UAE. This involves restructuring their entire regional team by employing management staff from countries such as Lebanon, Palestine, Iraq, and their labour force from the sub-continent such as India and Pakistan. This is not only due to the cheap wage rate but also to the ability to cope and work in such extreme hot weather conditions. The experience of individuals working in the region allows for future planning, where the difference in labour productivity during the extreme hot weather conditions is known, allowing precautionary measures to be put in place.

Construction and experimental study of an elevation linear Fresnel reflector

This paper outlines a novel elevation linear Fresnel reflector (ELFR) and presents and validates theoretical models defining its thermal performance. To validate the models, a series of experiments were carried out for receiver temperatures in the range of 30-100 °C to measure the heat loss coefficient, gain in heat transfer fluid (HTF) temperature, thermal efficiency, and stagnation temperature. The heat loss coefficient was underestimated due to the model exclusion of collector end heat losses. The measured HTF temperature gains were found to have a good correlation to the model predictions - less than a 5% difference. In comparison to model predictions for the thermal efficiency and stagnation temperature, measured values had a difference of -39% to +31% and 22-38%, respectively. The difference between the measured and predicted values was attributed to the low-temperature region for the experiments. It was concluded that the theoretical models are suitable for examining linear Fresnel reflector (LFR) systems and can be adopted by other researchers.

Floating Wetlands in the Puget Lowlands: Design, Construction, and Viability

Thesis (Master's)--University of Washington, 2016-06