Neomycin resistance as a dominant selectable marker for selection and isolation of vaccinia virus recombinants

The antibiotic G418 was shown to be an effective inhibitor of vaccinia virus replication when an appropriate concentration of it was added to cell monolayers 48 h before infection. Genetic engineering techniques were used in concert with DNA transfection protocols to construct vaccinia virus recombinants containing the neomycin resistance gene (neo) from transposon Tn5. These recombinants contained the neo gene linked in either the correct or incorrect orientation relative to the vaccinia virus 7.5-kilodalton gene promoter which is expressed constitutively throughout the course of infection. The vaccinia virus recombinant containing the chimeric neo gene in the proper orientation was able to grow and form plaques in the presence of G418, whereas both the wild-type and the recombinant virus with the neo gene in the opposite polarity were inhibited by more than 98%. The effect of G418 on virus growth may be mediated at least in part by selective inhibition of the synthesis of a subset of late viral proteins. These results are discussed with reference to using this system, the conferral of resistance to G418 with neo as a positive selectable marker, to facilitate constructing vaccinia virus recombinants which contain foreign genes of interest

The effect of spatial position of calorie information on choice, consumption and attention. ESRI WP615, February 2019

We report a “lab-in-the-field” experiment designed to test the impact of posting calories on menus. The study adds substantially to previous work by testing different spatial arrangements of price and calorie information. Choices were real, not hypothetical, and participants were unaware that their lunch choice was part of a study, even though their eye-movements were being tracked. Participants exposed to calorie information ordered 93 fewer calories (11%) relative to a control group. The impact was strongest when calorie information was presented on menus just to the right of the price, in an equivalent font. The difference in number of calories consumed was greater still. These effects were mediated by knowledge of the amount of calories in the meal, implying that calorie posting led to more informed decision making. There was no impact on enjoyment of the meal. Eye-tracking data suggested that this arrangement altered the decision process such that greater decision weight was given to calorie content

Expression of Sindbis virus structural proteins via recombinant vaccinia virus: synthesis, processing, and incorporation into mature Sindbis virions

We have obtained a vaccinia virus recombinant which contains a complete cDNA copy of the 26S RNA of Sindbis virus within the thymidine kinase gene of the vaccinia virus genome. This recombinant constitutively transcribed the Sindbis sequences throughout the infectious cycle, reflecting the dual early-late vaccinia promoter used in this construction. The Sindbis-derived transcripts were translationally active, giving rise to both precursor and mature structural proteins of Sindbis virus, including the capsid protein (C), the precursor of glycoprotein E2 (PE2), and the two mature envelope glycoproteins (E1 and E2). These are the same products translated from the 26S mRNA during Sindbis infection, and thus these proteins were apparently cleaved, glycosylated, and transported in a manner analogous to that seen during authentic Sindbis infections. By using epitope-specific antibodies, it was possible to demonstrate that recombinant-derived proteins were incorporated into Sindbis virions during coinfections with monoclonal antibody-resistant Sindbis variants. These results suggest that all the information necessary to specify the proper biogenesis of Sindbis virus structural proteins resides within the 26S sequences and that vaccinia may provide an appropriate system for using DNA molecular genetic manipulations to unravel a variety of questions pertinent to RNA virus replication

Human fur gene encodes a yeast KEX2-like endoprotease that cleaves pro-beta-NGF in vivo.

Extracts from BSC-40 cells infected with vaccinia recombinants expressing either the yeast KEX2 prohormone endoprotease or a human structural homologue (fur gene product) contained an elevated level of a membrane-associated endoproteolytic activity that could cleave at pairs of basic amino acids (-LysArg- and -ArgArg-). The fur-directed activity (furin) shared many properties with Kex2p including activity at pH 7.3 and a requirement for calcium. By using antifurin antibodies, immunoblot analysis detected two furin translation products (90 and 96 kD), while immunofluorescence indicated localization to the Golgi apparatus. Coexpression of either Kex2p or furin with the mouse beta-nerve growth factor precursor (pro-beta-NGF) resulted in greatly enhanced conversion of the precursor to mature nerve growth factor. Thus, the sequence homology shared by furin and the yeast KEX2 prohormone processing enzyme is reflected by significant functional homology both in vitro and in vivo

Preliminary Canopy Removal Experiments in Algal Dominated Communities Low on the Shore and in the Shallow Subtidal on the Isle of Man

The algal dominated communities immediately above and below the low-water spring level on a moderately exposed Manx shore were investigated by canopy removal experiments. Fucus serratus, Laminaria digitata and L. hyperborea were removed. Competition was shown to be important in determining the zonation of L. digitata and the distribution along the wave exposure gradient of other species such as Alaria esculenta, Desmarestia aculeata and D. viridis, and L. saccharina. Many species of algal epiphytes were early colonizers of canopy removal areas suggesting that competition from canopy algae usually restricts them to an epiphytic habit. The results indicate that interactions between macrophytes are much more important than grazing in structuring these communities

Thermoresponsive Gels based on ABC Triblock Copolymers: Effect of the Length of the PEG Side Group

ABC triblock copolymers of varying compositions and lengths of the PEG side groups were fabricated and their thermoresponsive behaviour was thoroughly investigated.</p

Spin Readout Techniques of the Nitrogen-Vacancy Center in Diamond

The diamond nitrogen-vacancy (NV) center is a leading platform for quantum information science due to its optical addressability and room-temperature spin coherence. However, measurements of the NV center's spin state typically require averaging over many cycles to overcome noise. Here, we review several approaches to improve the readout performance and highlight future avenues of research that could enable single-shot electron-spin readout at room temperature.Comment: 21 pages, 7 figure