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  • 1
    Online Resource
    Online Resource
    Multiplex Sequencing of 1.5 Mb of the Mycobacterium leprae  Genome
    Cold Spring Harbor Laboratory ; 1997
    In:  Genome Research Vol. 7, No. 8 ( 1997-08-01), p. 802-819
    Details
    In: Genome Research, Cold Spring Harbor Laboratory, Vol. 7, No. 8 ( 1997-08-01), p. 802-819
    Abstract: The nucleotide sequence of 1.5 Mb of genomic DNA from Mycobacterium leprae was determined using computer-assisted multiplex sequencing technology. This brings the 2.8-Mb M. leprae genome sequence to ∼66% completion. The sequences, derived from 43 recombinant cosmids, contain 1046 putative protein-coding genes, 44 repetitive regions, 3 rRNAs, and 15 tRNAs. The gene density of one per 1.4 kb is slightly lower than that of Mycoplasma (1.2 kb). Of the protein coding genes, 44% have significant matches to genes with well-defined functions. Comparison of 1157 M. leprae and 1564 Mycobacterium tuberculosis proteins shows a complex mosaic of homologous genomic blocks with up to 22 adjacent proteins in conserved map order. Matches to known enzymatic, antigenic, membrane, cell wall, cell division, multidrug resistance, and virulence proteins suggest therapeutic and vaccine targets. Unusual features of the M. leprae genome include large polyketide synthase (pks) operons, inteins, and highly fragmented pseudogenes. [The sequence data described in this paper have been submitted to GenBank under accession nos. L78811 – L78829 , U00010 – U00023 , U15180 – U15184 , U15186 , U15187 , L01095 , L01536 , L04666 , and L01263 . On-line supplementary information for Table 1 is available at http://www.cshl.org/gr .]
    Type of Medium: Online Resource
    ISSN: 1088-9051 , 1549-5469
    URL: Article
    DOI: 10.1101/gr.7.8.802
    RVK:
    XA 10000
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 1997
    detail.hit.zdb_id: 1483456-X
    SSG: 12
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  • 2
    Online Resource
    Online Resource
    An Efficient, Automatable Template Preparation for High Throughput Sequencing
    Mary Ann Liebert Inc ; 1998
    In:  Microbial & Comparative Genomics Vol. 3, No. 4 ( 1998-01), p. 237-241
    Details
    In: Microbial & Comparative Genomics, Mary Ann Liebert Inc, Vol. 3, No. 4 ( 1998-01), p. 237-241
    Type of Medium: Online Resource
    ISSN: 1090-6592
    URL: Article
    DOI: 10.1089/omi.1.1998.3.237
    Language: English
    Publisher: Mary Ann Liebert Inc
    Publication Date: 1998
    detail.hit.zdb_id: 2030312-9
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  • 3
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    Online Resource
    A Chemical Genetic Analysis of Platelet Activation.
    American Society of Hematology ; 2009
    In:  Blood Vol. 114, No. 22 ( 2009-11-20), p. 4009-4009
    Details
    In: Blood, American Society of Hematology, Vol. 114, No. 22 ( 2009-11-20), p. 4009-4009
    Abstract: Abstract 4009 Poster Board III-945 Platelets are anucleate cells that are not amenable to traditional forward genetic analysis. In collaboration with the Broad Institute Probe Development Center, we have performed a chemical genetic analysis of platelet activation. Chemical genetics involves exposure of cells to a library of small molecules, identification of compounds that produce a phenotype of interest, and determination of the target of these small molecules. We have used an assay designed both to identify allosteric inhibitors of Protease Activated Receptor-1 (PAR1) and to find inhibitors that selectively target granule release. This assay monitors dense granule secretion mediated by SFLLRN, a PAR1-specific agonist, using a luciferase-based assay system to detect ADP/ATP release. For primary screening, over 300,000 compounds were assayed in duplicate using freshly outdated platelet-rich plasma supplied by several blood banks across the United States. Computational analyses of the primary data demonstrated that approximately 0.2% of compounds showed ≥50% inhibition relative to maximally inhibitory concentrations of the known antiplatelet agent, cilostazol. Secondary screening using 8-point dose response curves were performed on the 629 inhibitory compounds, 742 compounds with inconclusive activity (e.g., ambiguous duplicates in primary screening), and 213 structural analogs of active compounds. These assays identified 367 active compounds with IC50s ≤10 micromolar. Counter screening to exclude luciferase inhibitors demonstrated 137 small molecules that inhibited PAR1-mediated ATP/ADP release without significant inhibition of luciferase. Database mining using PubChem and CAS search engines was performed to assess the selectivity of active compounds. Twenty eight compounds were selected for further testing based on their IC50s in confirmatory assays, lack of activity in unrelated bioassays, and chemical structure. Known platelet inhibitors were excluded. Of the 28 compounds, 16 compounds potently inhibited SFLLRN-induced alpha-granule release from washed platelets, as monitored by P-selectin expression. IC50s for these compounds ranged from 〈 0.3 to 1 micromolar. None of the selected compounds that failed to inhibit alpha-granule release demonstrated significant inhibition of SFLLRN-induced 14C-serotonin release at 10 micromolar. Ongoing studies are directed at selecting best candidates from among the 16 confirmed inhibitory compounds to develop as biological probes that target either PAR1 activation or distal steps in granule secretion. Disclosures: No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V114.22.4009.4009
    RVK:
    XA 33000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2009
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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