A Cell-Based Protein-Protein Interaction Method Using a Permuted Luciferase Reporter

We have developed a novel cell-based protein-protein interaction assay method. The method relies on conversion of an inactive permuted luciferase containing a Tobacco Etch Virus protease (TEV) cleavage sequence fused onto protein (A) to an active luciferase upon interaction and cleavage by another protein (B) fused with the TEV protease. We demonstrate assay applicability for ligand-induced protein-protein interactions including G-protein coupled receptors, receptor tyrosine kinases and nuclear hormone receptors

Properties of rat and mouse [beta]-glucuronidase mRNA and cDNA, including evidence for sequence polymorphism and genetic regulation of mRNA level

cDNA clones containing partial sequences for [beta]-glucuronidase ([beta]G) were constructed from rat preputial gland RNA and identified by their ability to selectively hybridize [beta]G mRNA. One such rat clone was used to isolate several cross-hybridizing clones from a mouse-cDNA library prepared from kidney RNA from androgen-treated animals. Together, the set of mouse clones spans about 2.0 kb of the 2.6-kb [beta]G mRNA. Using these cDNA clones as probes, a genomic polymorphism for DNA restriction fragment size was found that proved to be genetically linked to the [beta]G gene complex. A fragment of [beta]G cDNA was subcloned into a vector carrying an SP6 polymerase promoter to provide a template for the in vitro synthesis of single-stranded RNA complementary to [beta]G mRNA. This provided an extremely sensitive probe for the assay of [beta]G mRNA sequences. Using either nick-translated cDNA or transcribed RNA as a hybridization probe, we found that mouse [beta]G RNA levels are strongly induced by testosterone, and that induction by testosterone is pituitarydependent. During the lag period preceding induction, during the induction period itself, and during deinduction following removal of testosterone, [beta]G mRNA levels paralleled rates of [beta]G synthesis previously measured by in vivo pulse-labelling experiments. Genetic variation in the extent of induction affected either the level of [beta]G mRNA or its efficiency of translation depending on the strain of mice tested.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/25792/1/0000354.pd

Hudson Yards for Whom: Analysis of the Contribution of Hudson Yards in High-quality Public Space to New Yorkers

This thesis examines the extent to which Hudson Yards, a large-scale mixed-use development in New York City (NYC), fulfills its commitment to providing high-quality public spaces for New Yorkers. The study focuses on three main aspects: accessibility, diversity, and place identity. Employing a mixed-method approach, including site observation, GIS service area analysis, site plan analysis, tweets analysis, and an examination of historical public events and programs analysis, the study found that the Backyard at Hudson Yards does not provide sufficient accessibility design for people with disabilities. Although the extension of the 7 subway line has made the site relatively accessible to the general public through public transit, the site does not offer an engaging environment that encourages frequent, recurring visits. The place identity is primarily tourism-centric, as most discussions about the site revolve around landmarks. In conclusion, the Hudson Yards development has not achieved success in providing high-quality public space. This study contributes to the urban planning literature on high-quality public space, large-scale private development, and privately owned public space development (POPS) with a case study. The findings of this study emphasize the need for more inclusive design and planning practices in private developments, as well as the importantce of understanding the implications of such developments on various stakeholders

A recombinant capripoxvirus expressing the F protein of peste des petits ruminants virus and the P12A3C of foot-and-mouth disease virus

Abstract Background Peste des petits ruminants (PPR), foot-and-mouth disease (FMD) and sheep pox and goat pox are three important infectious diseases that infect goats, sheep and other small ruminants. It is well-known that the prevention of three diseases rely mainly on their individual vaccines. However, the vaccines have a variety of different disadvantages, such as short duration of immunity, increasing the number of vaccinations, and poor thermal stability. The purpose of this study is to construct a recombinant goat pox virus (rGPV) capable of expressing the F gene of PPRV and the P12A3C gene of FMDV as a live vector vaccine. Results The IRES, FMDV P12A3C and PPRV F genes into the multi-cloning site of the universal transfer plasmid pTKfpgigp to construct a recombinant transfer plasmid pTKfpgigpFiP12A3C, and transfected GPV-infected lamb testis (LT) cells with liposomes and produced by homologous recombination Recombinant GPV (rGPV/PPRVF-FMDVP12A3C, rGPV). The rGPV was screened and purified by green florescence protein (GFP) and xanthine-guanine-phosphoribosyltransferase gene (gpt) of Escherichia coli as selective markers, and the expression of rGPV in LT cells was detected by RT-PCR and immunofluorescence techniques. The results showed that the virus strain rGPV/PPRVF-FMDVP12A3C containing FMDV P12A3C and PPRV F genes was obtained. The exogenous genes FMDV P12A3C and PPRV F contained in rGPV were normally transcribed and translated in LT cells, and the expression products could specifically react with PPRV and FMDV antiserum. Then, the rGPV was intradermally inoculated with goats, the animal experiments showed that rGPV/PPRVF-FMDVP12A3C could induce high levels of specific antibodies against GPV, PPRV and FMDV. Conclusions The constructed rGPV induced high levels of specific antibodies against GPV, PPRV and FMDV. The study provides a reference for “ one vaccine with multiple uses “ of GPV live vector vaccine

Bulbophyllum reflexipetalum (Orchidaceae, Epidendroideae, Malaxideae), a new species from Xizang, China

Bulbophyllum reflexipetalum, a new species from Motuo County, Southeast Xizang, China, is described and illustrated here. This new species belongs to Bulbophyllum sect. Umbellata Bentham & J. D. Hooker, and it is morphologically similar to B. umbellatum Lindley, B. guttulatum (J. D. Hooker) N. P. Balakrishnan and B. salweenensis X.H. Jin, but is distinguished from them by having reflexed petals, base of dorsal sepal with 1 dentate on each side, lip with significantly revolute margin, adaxially with dark brown spots or patches and one longitudinal groove

Scalable Preparation of the Chemically Ordered Pt–Fe–Au Nanocatalysts with High Catalytic Reactivity and Stability for Oxygen Reduction Reactions

Carbon-supported Au–Pt<i>_x</i>Fe<i>_y</i> nanoparticles were synthesized via microwave heating polyol process, followed by annealing for the formation of the ordered structure. The structure characterizations indicate that Au is alloyed with intermetallic Pt–Fe nanoparticles and therefore the surface electronic properties are tuned. The electrochemical tests show that the microwave heating polyol process is more effective than oil bath heating polyol process for synthesizing the highly active catalysts. The introduction of trace Au (0.2 wt % Au) significantly improves the oxygen reduction reaction (ORR) catalytic activity of Pt<i>_x</i>Fe<i>_y</i> catalysts. Au–PtFe/C–H (0.66 A/mg_Pt) and Au–PtFe₃/C–H (0.63 A/mg_Pt) prepared in a batch of 10.0 g show significantly improved catalytic activities than their counterparts (PtFe/C–H and PtFe₃/C–H) as well as commercial Johnson Matthey Pt/C (0.17 A/mg_Pt). In addition, the as-prepared Au–PtFe/C–H and Au–PtFe₃/C–H display highly enhanced stability toward the ORR compared to the commercial Pt/C. The superior catalytic performance is attributed to the synergistic effect of chemically ordered intermetallic structure and Au. This work provides a scalable synthesis of the multimetallic chemically ordered Au–Pt<i>_x</i>Fe<i>_y</i> catalysts with high ORR catalytic performance in acidic condition