187 research outputs found

    Microcollinearity in an ethylene receptor coding gene region of the Coffea canephora genome is extensively conserved with Vitis vinifera and other distant dicotyledonous sequenced genomes

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    Background: Coffea canephora, also called Robusta, belongs to the Rubiaceae, the fourth largest angiosperm family. This diploid species (2x = 2n = 22) has a fairly small genome size of approximate to 690 Mb and despite its extreme economic importance, particularly for developing countries, knowledge on the genome composition, structure and evolution remain very limited. Here, we report the 160 kb of the first C. canephora Bacterial Artificial Chromosome (BAC) clone ever sequenced and its fine analysis. Results: This clone contains the CcEIN4 gene, encoding an ethylene receptor, and twenty other predicted genes showing a high gene density of one gene per 7.8 kb. Most of them display perfect matches with C. canephora expressed sequence tags or show transcriptional activities through PCR amplifications on cDNA libraries. Twenty-three transposable elements, mainly Class II transposon derivatives, were identified at this locus. Most of these Class II elements are Miniature Inverted-repeat Transposable Elements (MITE) known to be closely associated with plant genes. This BAC composition gives a pattern similar to those found in gene rich regions of Solanum lycopersicum and Medicago truncatula genomes indicating that the CcEIN4 regions may belong to a gene rich region in the C. canephora genome. Comparative sequence analysis indicated an extensive conservation between C. canephora and most of the reference dicotyledonous genomes studied in this work, such as tomato (S. lycopersicum), grapevine (V. vinifera), barrel medic M. truncatula, black cottonwood (Populus trichocarpa) and Arabidopsis thaliana. The higher degree of microcollinearity was found between C. canephora and V. vinifera, which belong respectively to the Asterids and Rosids, two clades that diverged more than 114 million years ago. Conclusion: This study provides a first glimpse of C. canephora genome composition and evolution. Our data revealed a remarkable conservation of the microcollinearity between C. canephora and V. vinifera and a high conservation with other distant dicotyledonous reference genomes. Altogether, these results provide valuable information to identify candidate genes in C. canephora genome and serve as a foundation to establish strategies for whole genome sequencing. Future large-scale sequence comparison between C. canephora and reference sequenced genomes will help in understanding the evolutionary history of dicotyledonous plants

    Patterns of sequence polymorphism in the fleshless berry locus in cultivated and wild Vitis vinifera accessions

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    <p>Abstract</p> <p>Background</p> <p>Unlike in tomato, little is known about the genetic and molecular control of fleshy fruit development of perennial fruit trees like grapevine (<it>Vitis vinifera </it>L.). Here we present the study of the sequence polymorphism in a 1 Mb grapevine genome region at the top of chromosome 18 carrying the <it>fleshless berry </it>mutation (<it>flb</it>) in order, first to identify SNP markers closely linked to the gene and second to search for possible signatures of domestication.</p> <p>Results</p> <p>In total, 62 regions (17 SSR, 3 SNP, 1 CAPS and 41 re-sequenced gene fragments) were scanned for polymorphism along a 3.4 Mb interval (85,127-3,506,060 bp) at the top of the chromosome 18, in both <it>V. vinifera cv</it>. Chardonnay and a genotype carrying the <it>flb </it>mutation, <it>V. vinifera cv</it>. Ugni Blanc mutant. A nearly complete homozygosity in Ugni Blanc (wild and mutant forms) and an expected high level of heterozygosity in Chardonnay were revealed. Experiments using qPCR and BAC FISH confirmed the observed homozygosity. Under the assumption that <it>flb </it>could be one of the genes involved into the domestication syndrome of grapevine, we sequenced 69 gene fragments, spread over the <it>flb </it>region, representing 48,874 bp in a highly diverse set of cultivated and wild <it>V. vinifera </it>genotypes, to identify possible signatures of domestication in the cultivated <it>V. vinifera </it>compartment. We identified eight gene fragments presenting a significant deviation from neutrality of the Tajima's D parameter in the cultivated pool. One of these also showed higher nucleotide diversity in the wild compartments than in the cultivated compartments. In addition, SNPs significantly associated to berry weight variation were identified in the <it>flb </it>region.</p> <p>Conclusions</p> <p>We observed the occurrence of a large homozygous region in a non-repetitive region of the grapevine otherwise highly-heterozygous genome and propose a hypothesis for its formation. We demonstrated the feasibility to apply BAC FISH on the very small grapevine chromosomes and provided a specific probe for the identification of chromosome 18 on a cytogenetic map. We evidenced genes showing putative signatures of selection and SNPs significantly associated with berry weight variation in the <it>flb </it>region. In addition, we provided to the community 554 SNPs at the top of chromosome 18 for the development of a genotyping chip for future fine mapping of the <it>flb </it>gene in a F2 population when available.</p

    Development of a prototype superconducting radio-frequency cavity for conduction-cooled accelerators

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    The higher efficiency of superconducting radio-frequency (SRF) cavities compared to normal-conducting ones enables the development of high-energy continuous-wave linear accelerators (linacs). Recent progress in the development of high-quality Nb3_3Sn film coatings along with the availability of cryocoolers with high cooling capacity at 4 K makes it feasible to operate SRF cavities cooled by thermal conduction at relevant accelerating gradients for use in accelerators. A possible use of conduction-cooled SRF linacs is for environmental applications, requiring electron beams with energy of 1101 - 10 MeV and 1 MW of power. We have designed a 915 MHz SRF linac for such an application and developed a prototype single-cell cavity to prove the proposed design by operating it with cryocoolers at the accelerating gradient required for 1 MeV energy gain. The cavity has a 3\sim 3 μ\mum thick Nb3_3Sn film on the inner surface, deposited on a 4\sim4 mm thick bulk Nb substrate and a bulk 7\sim7 mm thick Cu outer shell with three Cu attachment tabs. The cavity was tested up to a peak surface magnetic field of 53 mT in liquid He at 4.3 K. A horizontal test cryostat was designed and built to test the cavity cooled with three Gifford-McMahon cryocoolers. The rf tests of the conduction-cooled cavity, performed at General Atomics, achieved a peak surface magnetic field of 50 mT and stable operation was possible with up to 18.5 W of rf heat load. The peak frequency shift due to microphonics was 23 Hz. These results represent the highest peak surface magnetic field achieved in a conduction-cooled SRF cavity to date and meet the requirements for a 1 MeV energy gain

    Mouse Dfa Is a Repressor of TATA-box Promoters and Interacts with the Abt1 Activator of Basal Transcription

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    Our study of the mouse Ate1 arginyltransferase, a component of the N-end rule pathway, has shown that Ate1 pre-mRNA is produced from a bidirectional promoter that also expresses, in the opposite direction, a previously uncharacterized gene (Hu, R. G., Brower, C. S., Wang, H., Davydov, I. V., Sheng, J., Zhou, J., Kwon, Y. T., and Varshavsky, A. (2006) J. Biol. Chem. 281, 32559–32573). In this work, we began analyzing this gene, termed Dfa (divergent from Ate1). Mouse Dfa was found to be transcribed from both the bidirectional P_(Ate1/Dfa) promoter and other nearby promoters. The resulting transcripts are alternatively spliced, yielding a complex set of Dfa mRNAs that are present largely, although not exclusively, in the testis. A specific Dfa mRNA encodes, via its 3′-terminal exon, a 217-residue protein termed Dfa^A. Other Dfa mRNAs also contain this exon. DfaA is sequelogous (similar in sequence) to a region of the human/mouse HTEX4 protein, whose physiological function is unknown. We produced an affinity-purified antibody to recombinant mouse DfaA that detected a 35-kDa protein in the mouse testis and in several cell lines. Experiments in which RNA interference was used to down-regulate Dfa indicated that the 35-kDa protein was indeed Dfa^A. Furthermore, Dfa^A was present in the interchromatin granule clusters and was also found to bind to the Ggnbp1 gametogenetin-binding protein-1 and to the Abt1 activator of basal transcription that interacts with the TATA-binding protein. Given these results, RNA interference was used to probe the influence of Dfa levels in luciferase reporter assays. We found that Dfa^A acts as a repressor of TATA-box transcriptional promoters

    An application of tomographic PIV to investigate the spray-induced turbulence in a direct-injection engine

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    Fuel sprays produce high-velocity, jet-like flows that impart turbulence onto the ambient flow field. The spray-induced turbulence augments fuel-air mixing, which has a primary role in controlling pollutant formation and cyclic variability in engines. This paper presents tomographic particle image velocimetry (TPIV) measurements to analyse the 3D spray-induced turbulence during the intake stroke of a direct-injection engine. The spray produces a strong spray-induced jet in the far field, which travels through the cylinder and imparts turbulence onto the surrounding flow. Planar high-speed PIV measurements at 4.8 kHz are combined with TPIV at 3.3 Hz to evaluate spray particle distributions and validate TPIV measurements in the particle-laden flow. An uncertainty analysis is performed to assess the uncertainty associated with vorticity and strain rate components. TPIV analyses quantify the spatial domain of the turbulence in relation to the SIJ and describe how turbulent flow features such as turbulent kinetic energy, strain rate and vorticity evolve into the surrounding flow field. Access to the full tensors facilitate the evaluation of turbulence for individual spray events. TPIV images reveal the presence of strong shear layers (visualized by high S magnitudes) and pockets of elevated vorticity along the immediate boundary of the SIJ. Values are extracted from spatial domains extending in 1mm increments from the SIJ. Turbulence levels are greatest within the 0-1mm region from the SIJ boarder and dissipate with radial distance. Individual strain rate and vorticity components are analyzed in detail to describe the relationship between local strain rates and 3D vortical structures produced within strong shear layers of the SIJ. Analyses are intended to understand the flow features responsible for rapid fuel-air mixing and provide valuable data for the development of numerical models

    Integrative analysis of the human cis-antisense gene pairs, miRNAs and their transcription regulation patterns

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    Cis-antisense gene pairs (CASGPs) can transcribe mRNAs from an opposite strand of a given locus. To classify and understand diverse CASGP phenomena in the human we compiled a genome-wide catalog of CASGPs and integrated these sequences with microarray, SAGE and miRNA data. Using the concept of overlapping regions and clustering of SA transcripts by chromosome coordinates, we identified up to 9000 overlapping antisense loci. Four thousand three hundred and seventy-four of these CASGPs form 1759 complex gene architectures. We found that ∼35% (6347/18160) of RefSeq genes are overlapped with the antisense transcripts. About 30% of Affymetrix U133 microarray initial sequences map transcripts of ∼35% CASGPs and reveal mostly concordant expression in CASGPs. We found strong significant overrepresentation of human miRNA genes in loci of CASGPs. We developed a data-driven model of cross-talk between co-expressed CASGPs and DICER1-mediated miRNA pathway in normal spermatogenesis and in severe teratozoospermia. Specifically, we revealed complex SA structural–functional gene module composing the protein-coding genes, WDR6, DALRD3, NDUFAF3 and ncRNA precursors, mir-425 and mir-191, which could provide downregulation of ncRNA pathway via direct targeting DICER1 and basonuclin 2 transcripts by mir-425 and mir-191 in normal spermatogenesis, but this mechanism is switched off in severe teratozoospermia. The database is available from http://globalisland.bii.a-star.edu.sg/∼jiangtao/sas/index3.php?link =abou
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