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.

Evaluation of genetic markers as instruments for Mendelian randomization studies on vitamin D

Combining SNPs into allele scores provides a more powerful instrument for MR analysis than a single SNP in isolation. Population stratification and the potential for pleiotropic effects need to be considered in MR studies on vitamin D.

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 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.

Samuel Beckett's digital library

A digital reconstruction of Samuel Beckett's personal library, based on the volumes preserved at his apartment in Paris, in archives (Beckett International Foundation) and private collections (James and Elizabeth Knowlson Collection, Anne Atik, Noga Arikha, Terrence Killeen,...).

Sequence and function of lysosomal and digestive cathepsin D-like proteinases of Musca domestica midgut

Musca domestica larvae display in anterior and middle midgut contents, a proteolytic activity with pH optimum of 3.0-3.5 and kinetic properties like cathepsin D. Three cDNAs coding for preprocathepsin D-like proteinases (ppCAD 1, ppCAD 2, ppCAD 3) were cloned from a M. domestica midgut cDNA library. The coded protein sequences included the signal peptide, propeptide and mature enzyme that has all conserved catalytic and substrate binding residues found in bovine lysosomal cathepsin D. Nevertheless, ppCAD 2 and ppCAD 3 lack the characteristic proline loop and glycosylation sites. A comparison among the sequences of cathepsin D-like enzymes from some vertebrates and those found in M. domestica and in the genomes of Aedes aegypti, Drosophila melanogaster, Tribolium castaneum, and Bombyx mori showed that only flies have enzymes lacking the proline loop (as defined by the motif: DxPxPx(G/A)P), thus resembling vertebrate pepsin. ppCAD 3 should correspond to the digestive cathepsin D-like proteinase (CAD) found in enzyme assays because: (1) it seems to be the most expressed CAD, based on the frequency of ESTs found. (2) The mRNA for CAD 3 is expressed only in the anterior and proximal middle midgut. (3) Recombinant procathepsin D-like proteinase (pCAD 3), after auto-activation has a pH optimum of 2.5-3.0 that is close to the luminal pH of M. domestica midgut. (4) Immunoblots of proteins from different tissues revealed with anti-pCAD 3 serum were positive only in samples of anterior and middle midgut tissue and contents. (5) CAD 3 is localized with immunogold inside secretory vesicles and around microvilli in anterior and middle midguit cells. The data support the view that on adapting to deal with a bacteria-rich food in an acid midgut region, M. domestica digestive CAD resulted from the same archetypical gene as the intracellular cathepsin D, paralleling what happened with vertebrates. The lack of the proline loop may be somehow associated with the extracellular role of both pepsin and digestive CAD 3. (C) 2009 Elsevier Ltd. All rights reserved.

Graduate School and University Center Archives Finding Aid - Record Group V-D: Presidents' Files: Kelly

This is part of the finding aid to the Graduate School and University Center (GSUC) Archives. Record Group V-D is the papers of William P. Kelly from when he was president of the GSUC (2005-2013).

O bibliotecário perfeito: o historiador Ramiz Galvão na Biblioteca Nacional

Em 1870, o jovem e praticamente desconhecido Benjamin Franklin Ramiz Galvão foi nomeado para ocupar o cargo de diretor da Biblioteca Nacional, posto do qual saiu doze anos depois, já como intelectual consagrado, para assumir a função de tutor dos netos de D. Pedro II. Durante o período em que esteve à frente desta instituição, Ramiz Galvão projetou fazer daquele espaço a biblioteca da nação brasileira. Isso significava construir uma instituição cuja função fosse salvaguardar o patrimônio documental brasileiro, torná-lo disponível a um público especializado e estimular estudos sobre a história e a geografia do país. O objetivo era fazer da Biblioteca Nacional um lugar de referência, sintonizada com suas congêneres europeias e em diálogo com os meios letrados nacionais. Nesse sentido, a atuação de Galvão foi fundamental para a construção de uma rede de sociabilidade que o ligava a estudiosos, livreiros, bibliófilos e bibliotecários de diversos lugares do Brasil, da América e da Europa. Esta tese analisa a proposta de reformulação da Biblioteca Nacional empreendida por Ramiz Galvão, buscando associá-la a um projeto maior de construção da nação brasileira. Para isso, investigaremos três elementos que consideramos principais desse trabalho de edificação de uma Biblioteca nacional: 1) a construção de uma rotina de serviços, que foi capaz de transformar o pequeno espaço com livros da antiga Biblioteca Real na mais importante biblioteca do Brasil; 2) o esforço de aquisição e de seleção de um patrimônio documental brasileiro e sua divulgação em publicações como os Anais da Biblioteca Nacional e o Catálogo da Exposição de História do Brasil, dois de seus mais importantes empreendimentos como diretor; 3) a atuação de Ramiz Galvão como mediador cultural no ambiente letrado brasileiro do final do século XIX.

Development and characterization of 14 microsatellite loci from an enriched genomic library of Eucalyptus camaldulensis Dehnh

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

SMase II, a new sphingomyelinase D from Loxosceles laeta venom gland: Molecular cloning, expression, function and structural analysis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Molecular characterization of hemoglobin alpha-D chains from Geochelone carbonaria and Geochelone denticulata land turtles

In order to help elucidate the evolution of alpha-globins, the complete cDNA and amino acid sequences of Geochelone carbonaria and Geochelone denticulata land turtles alpha-D chains have been described. In G. carbonaria, the cDNA is 539 bp with ATG start codon located at position 46, TGA stop codon at position 469 and AATAAA polyadenylation signal at position 520. In G. denticulata, the cDNA is 536 bp with ATG start codon located at position 46, TGA stop codon at position 469 and AATAAA polyadenylation signal at position 517. Both cDNAs codify 141 amino acid residues, differing from each other in only four amino acid residues. When comparing with human Hb alpha-chain, alterations in important regions can be noted: alpha110 Ala-Gly, alpha114 Pro-Gly, alpha117 Phe-Tyr and alpha122 His-Gln. There is a high homology between the amino acids of these turtles when compared with chicken alpha-D chains, progressively decreasing when compared with human, crocodile, snake, frog and fish alpha-chains. Phylogenetic analysis of alpha-D chains shows that those of turtles are closer to those of birds than to snakes and lizards. (C) 2002 Elsevier B.V. All rights reserved.

Vitamin D Induces Increased Systolic Arterial Pressure via Vascular Reactivity and Mechanical Properties

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)