STUDY OF A SINGLE-CHARGED IONS ECR SOURCE MATCHING OF THE EXTRACTED BEAM TO AN ISOTOPE SEPARATOR

A new ECR ion-source has been designed and studied for single-charged ion beams. A very stable regime has been obtained with an ion-source made of two identical stages in cascade. The RF power supplies consist of two 2.45 GHz magnetrons. The discharge chamber is made of two coaxial Pyrex tubes. The external one ensures vacuum and HT insulation. The tubes are aligned inside the two multimode cavities axially limited by three magnetic coils. The ion beam is extracted at 20 kV and focused with electric lenses. For argon and xenon, 1 mA single-charged ion currents have been extracted. The influence of various parameters has been progressively achieved with a set-up including a 60° analyzing magnet and with the 120° on-line isotope separator at SARA. From emittances and images observed it appears difficult to compensate charge space effects. Suggestions and future developments are proposed to improve qualities of the isotopic separation

Helicobacter pylori Type IV Secretion Apparatus Exploits β1 Integrin in a Novel RGD-Independent Manner

Translocation of the Helicobacter pylori (Hp) cytotoxin-associated gene A (CagA) effector protein via the cag-Type IV Secretion System (T4SS) into host cells is a major risk factor for severe gastric diseases, including gastric cancer. However, the mechanism of translocation and the requirements from the host cell for that event are not well understood. The T4SS consists of inner- and outer membrane-spanning Cag protein complexes and a surface-located pilus. Previously an arginine-glycine-aspartate (RGD)-dependent typical integrin/ligand type interaction of CagL with α5β1 integrin was reported to be essential for CagA translocation. Here we report a specific binding of the T4SS-pilus-associated components CagY and the effector protein CagA to the host cell β1 Integrin receptor. Surface plasmon resonance measurements revealed that CagA binding to α5β1 integrin is rather strong (dissociation constant, KD of 0.15 nM), in comparison to the reported RGD-dependent integrin/fibronectin interaction (KD of 15 nM). For CagA translocation the extracellular part of the β1 integrin subunit is necessary, but not its cytoplasmic domain, nor downstream signalling via integrin-linked kinase. A set of β1 integrin-specific monoclonal antibodies directed against various defined β1 integrin epitopes, such as the PSI, the I-like, the EGF or the β-tail domain, were unable to interfere with CagA translocation. However, a specific antibody (9EG7), which stabilises the open active conformation of β1 integrin heterodimers, efficiently blocked CagA translocation. Our data support a novel model in which the cag-T4SS exploits the β1 integrin receptor by an RGD-independent interaction that involves a conformational switch from the open (extended) to the closed (bent) conformation, to initiate effector protein translocation

FLP Recombinase-Mediated Site-Specific Recombination in Silkworm, Bombyx mori

A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they have not yet been established for use in the manipulation of the silkworm Bombyx mori genome. In this study, we achieved site-specific excision of a target gene at predefined chromosomal sites in the silkworm using a FLP/FRT site-specific recombination system. We first constructed two stable transgenic target silkworm strains that both contain a single copy of the transgene construct comprising a target gene expression cassette flanked by FRT sites. Using pre-blastoderm microinjection of a FLP recombinase helper expression vector, 32 G3 site-specific recombinant transgenic individuals were isolated from five of 143 broods. The average frequency of FLP recombinase-mediated site-specific excision in the two target strains genome was approximately 3.5%. This study shows that it is feasible to achieve site-specific recombination in silkworms using the FLP/FRT system. We conclude that the FLP/FRT system is a useful tool for genome manipulation in the silkworm. Furthermore, this is the first reported use of the FLP/FRT system for the genetic manipulation of a lepidopteran genome and thus provides a useful reference for the establishment of genome manipulation technologies in other lepidopteran species

STUDY OF A SINGLE-CHARGED IONS ECR SOURCE MATCHING OF THE EXTRACTED BEAM TO AN ISOTOPE SEPARATOR

Une nouvelle source ECR a été étudiée pour la production d'ions monochargés. Un régime très stable a été obtenu avec une source d'ions à deux étages identiques en cascade. La puissance RF est fournie à l'aide de deux magnétrons à 2.45 GHz. La chambre a été réalisée à l'aide de deux tubes coaxiaux de Pyrex, le tube extérieur assurant l'étanchéité au vide et l'isolement électrique. Les tubes sont alignés à l'intérieur de deux cavités multimodes axialement limités par trois bobines magnétiques. Le faisceau d'ions est accéléré à 20 kV et focalisé à l'aide de lentilles électriques. Pour l'argon et le xénon, des courants de 1 mA d'ions monochargés ont été extraits. L'influence des divers paramètres de la source a été étudiée progressivement sur un ensemble comportant un aimant d'analyse de 60° et sur le séparateur d'isotopes 120° en ligne sur l'accélérateur SARA. Les émittances et les images observées ont montré les difficultés rencontrées pour compenser les effets de charge d'espace. Des suggestions et des développements futurs sont proposés pour obtenir des conditions de séparation isotopique convenables.A new ECR ion-source has been designed and studied for single-charged ion beams. A very stable regime has been obtained with an ion-source made of two identical stages in cascade. The RF power supplies consist of two 2.45 GHz magnetrons. The discharge chamber is made of two coaxial Pyrex tubes. The external one ensures vacuum and HT insulation. The tubes are aligned inside the two multimode cavities axially limited by three magnetic coils. The ion beam is extracted at 20 kV and focused with electric lenses. For argon and xenon, 1 mA single-charged ion currents have been extracted. The influence of various parameters has been progressively achieved with a set-up including a 60° analyzing magnet and with the 120° on-line isotope separator at SARA. From emittances and images observed it appears difficult to compensate charge space effects. Suggestions and future developments are proposed to improve qualities of the isotopic separation