Baculovirus superinfection: a probable restriction factor on the surface display of proteins for library screening

In addition to the expression of recombinant proteins, baculoviruses have been developed as a platform for the display of complex eukaryotic proteins on the surface of virus particles or infected insect cells. Surface display has been used extensively for antigen presentation and targeted gene delivery but is also a candidate for the display of protein libraries for molecular screening. However, although baculovirus gene libraries can be efficiently expressed and displayed on the surface of insect cells, target gene selection is inefficient probably due to super-infection which gives rise to cells expressing more than one protein. In this report baculovirus superinfection of Sf9 cells has been investigated by the use of two recombinant multiple nucleopolyhedrovirus carrying green or red fluorescent proteins under the control of both early and late promoters (vAcBacGFP and vAcBacDsRed). The reporter gene expression was detected 8 hours after the infection of vAcBacGFP and cells in early and late phases of infection could be distinguished by the fluorescence intensity of the expressed protein. Simultaneous infection with vAcBacGFP and vAcBacDsRed viruses each at 0.5 MOI resulted in 80% of infected cells coexpressing the two fluorescent proteins at 48 hours post infection (hpi), and subsequent infection with the two viruses resulted in similar co-infection rate. Most Sf9 cells were re-infectable within the first several hours post infection, but the reinfection rate then decreased to a very low level by 16 hpi. Our data demonstrate that Sf9 cells were easily super-infectable during baculovirus infection, and super-infection could occur simultaneously at the time of the primary infection or subsequently during secondary infection by progeny viruses. The efficiency of super-infection may explain the difficulties of baculovirus display library screening but would benefit the production of complex proteins requiring co-expression of multiple polypeptides.

Boots Book-lovers' Library and the novel: The impact of a circulating library market on twentieth-century fiction

In a Report for the Society of Bookmen in 1928, British publishers estimated that between a quarter to two thirds of all the books they published went to four circulating libraries: Boots, Smith’s, Mudie’s, and The Times bookclub. This essay examines the literary impact of one of the largest of these, Boots Book-lovers’ Library (1899-66), which by 1935 had around 400 libraries attached to their high-street pharmacies catering for the tastes of over one million subscribers a year. Compared to the wealth of studies examining the influence of the library market in the Victorian period, the significance of the subscription libraries as key distributors of fiction in the twentieth century is not well known. But private libraries expanded rapidly in the early twentieth century to cater for what Sidney Dark termed a ‘new reading public’, and records in publishers’ archives indicate that authors routinely adapted their unpublished manuscripts in order to meet the perceived demands of this library reader. This article examines the impact of the Boots Book-lovers’ Library market on authors’ practices of writing and revision, and on literary marketing and censorship. It focuses in particular on the author James Hanley (1897-1985), using unpublished correspondence in the Chatto & Windus archive at the University of Reading to demonstrate how the publisher’s sense of the tastes and expectations of the Boots library reader influenced the editorial process.

Construction of a Festuca pratensis BAC library for map-based cloning in Festulolium substitution lines

Introgression in Festulolium is a potentially powerful tool to isolate genes for a large number of traits which differ between Festuca pratensis Huds. and Lolium perenne L. Not only are hybrids between the two species fertile, but the two genomes can be distinguished by genomic in situ hybridisation and a high frequency of recombination occurs between homoeologous chromosomes and chromosome segments. By a programme of introgression and a series of backcrosses, L. perenne lines have been produced which contain small F. pratensis substitutions. This material is a rich source of polymorphic markers targeted towards any trait carried on the F. pratensis substitution not observed in the L. perenne background. We describe here the construction of an F. pratensis BAC library, which establishes the basis of a map-based cloning strategy in L. perenne. The library contains 49,152 clones, with an average insert size of 112 kbp, providing coverage of 2.5 haploid genome equivalents. We have screened the library for eight amplified fragment length polymorphism (AFLP) derived markers known to be linked to an F. pratensis gene introgressed into L. perenne and conferring a staygreen phenotype as a consequence of a mutation in primary chlorophyll catabolism. While for four of the markers it was possible to identify bacterial artificial chromosome (BAC) clones, the other four AFLPs were too repetitive to enable reliable identification of locus-specific BACs. Moreover, when the four BACs were partially sequenced, no obvious coding regions could be identified. This contrasted to BACs identified using cDNA sequences, when multiple genes were identified on the same BAC.

A maize bacterial artificial chromosome (BAC) library from the European flint inbred line F2

We report here the construction and characterisation of a BAC library from the maize flint inbred line F2, widely used in European maize breeding programs. The library contains 86,858 clones with an average insert size of approximately 90 kb, giving approximately 3.2-times genome coverage. High-efficiency BAC cloning was achieved through the use of a single size selection for the high-molecular-weight genomic DNA, and co-transformation of the ligation with yeast tRNA to optimise transformation efficiency. Characterisation of the library showed that less than 0.5% of the clones contained no inserts, while 5.52% of clones consisted of chloroplast DNA. The library was gridded onto 29 nylon filters in a double-spotted 8 × 8 array, and screened by hybridisation with a number of single-copy and gene-family probes. A 3-dimensional DNA pooling scheme was used to allow rapid PCR screening of the library based on primer pairs from simple sequence repeat (SSR) and expressed sequence tag (EST) markers. Positive clones were obtained in all hybridisation and PCR screens carried out so far. Six BAC clones, which hybridised to a portion of the cloned Rp1-D rust resistance gene, were further characterised and found to form contigs covering most of this complex resistance locus.

A genealogy of the kings of England in papal Avignon: British Library, Egerton MS. 1500

This is one of three articles in eBLJ (2013) dedicated to the "Abreujamen de las estorias", British Library Egerton MS 1500. The other two articles are: Federico Botana,' The Making of "L'Abreujamen de las estorias": British Library Egerton MS 1500', eblj (2013), article 16. Alexander Ibarz, 'The Provenance of the "Abreujamens de las estorias" (London, British Library, Egerton MS. 1500) and the Identification of Scribal Hands (c. 1323)' eblj (2013), article 17. Leglu's article, eblj (2013), article 18, examines the various political agendas that emerge in the depiction of the kings of Britain and England, from Brutus to Edward II.

Derivation of an urban materials spectral library through emittance and reflectance spectroscopy

Recent advances in thermal infrared remote sensing include the increased availability of airborne hyperspectral imagers (such as the Hyperspectral Thermal Emission Spectrometer, HyTES, or the Telops HyperCam and the Specim aisaOWL), and it is planned that an increased number spectral bands in the long-wave infrared (LWIR) region will soon be measured from space at reasonably high spatial resolution (by imagers such as HyspIRI). Detailed LWIR emissivity spectra are required to best interpret the observations from such systems. This includes the highly heterogeneous urban environment, whose construction materials are not yet particularly well represented in spectral libraries. Here, we present a new online spectral library of urban construction materials including LWIR emissivity spectra of 74 samples of impervious surfaces derived using measurements made by a portable Fourier Transform InfraRed (FTIR) spectrometer. FTIR emissivity measurements need to be carefully made, else they are prone to a series of errors relating to instrumental setup and radiometric calibration, which here relies on external blackbody sources. The performance of the laboratory-based emissivity measurement approach applied here, that in future can also be deployed in the field (e.g. to examine urban materials in situ), is evaluated herein. Our spectral library also contains matching short-wave (VIS–SWIR) reflectance spectra observed for each urban sample. This allows us to examine which characteristic (LWIR and) spectral signatures may in future best allow for the identification and discrimination of the various urban construction materials, that often overlap with respect to their chemical/mineralogical constituents. Hyperspectral or even strongly multi-spectral LWIR information appears especially useful, given that many urban materials are composed of minerals exhibiting notable reststrahlen/absorption effects in this spectral region. The final spectra and interpretations are included in the London Urban Micromet data Archive (LUMA; http://LondonClimate.info/LUMA/SLUM.html).

Sir Bernard Spilsbury: a survey and catalogue of his autopsy case cards from the Wellcome Library, London

During his lifetime, Sir Bernard Spilsbury was referred to as the ‘‘father of forensic medicine.’’ He became a household name as a result of several famous cases. Several articles have been written about his life and work, but an objective assessment has proved difficult because of the lack of available material that Spilsbury himself produced. His main legacy has been a series of case cards, but for many years these were unavailable to the researcher. In 2008, a collection of some 4000 of Spilsbury’s case cards was bought by The Wellcome Library in London and therefore entered the public domain. In this article, we report our study of 650 of these cards. We discuss trends in Spilsbury’s work and several specific cases in more detail. These cards allow an objective view to be taken of Spilsbury’s everyday work, and we feel that some reappraisal of his legacy is now timely

The Italian Academies, 1525-1700: a themed collection database, London, British Library

A database of books published by Italian Academies between 1525 and 1700. The Italian Academies Themed Collection provides a detailed searchable database for locating printed material relating to the Italian learned Academies active in Avellino, Bari, Benevento, Bologna, Brindisi, Caltanissetta, Catania, Catanzaro, Enna, L’Aquila, Lecce, Mantua, Naples, Padua, Palermo, Rome, Salerno, Siena, Syracuse, Trapani, and Venice in the period 1525-1700 and now held in the collections of the British Library.

The Environmental Data Abstraction Library (EDAL): a modular approach to processing and visualising large environmental data

The Environmental Data Abstraction Library provides a modular data management library for bringing new and diverse datatypes together for visualisation within numerous software packages, including the ncWMS viewing service, which already has very wide international uptake. The structure of EDAL is presented along with examples of its use to compare satellite, model and in situ data types within the same visualisation framework. We emphasize the value of this capability for cross calibration of datasets and evaluation of model products against observations, including preparation for data assimilation.

The Devil’s Daughters and a question of translation between Occitan and Anglo-Norman French: ‘De las .vii. filhas del diable’ (British Library Add. MS 17920).

The evidence for vernacular-to-vernacular translation is hard to demonstrate in medieval Romance languages. This article analyses a hypothesis published a century ago that there is an identifiable Anglo-Norman source for an Occitan prose text. Both texts spring from a Latin exemplum in which the seven capital vices are personified as the Devil's daughters, married off to seven social categories (the clergy, knights, peasants, etc.). Although the hypothesis is disproved, it remains that the dialogue between Anglo-Norman French and Occitan has been overlooked, and deserves further exploration.