Isolation and Characterization of Effector Genes in Pseudomonas Syringae PV. Tomato Strain DC3000 PDF Download
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Languages : en
Pages :
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Languages : en
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Author: Nikolaos Skandalis
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Category :
Languages : en
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Author: Kathy Roberts Munkvold
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Category :
Languages : en
Pages : 286
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Author: Julie Zwiesler-Vollick
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Category : Arabidopsis thaliana
Languages : en
Pages : 412
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Author: Sruti Bandyopadhyay
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Category : Plant-pathogen relationships
Languages : en
Pages : 396
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Author: Dennis P. Jackson
Publisher:
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Category : Pseudomonas syringae
Languages : en
Pages : 340
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Author: Vanessa Revindran
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Category : Biochemistry
Languages : en
Pages : 262
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We have created a yeast madel system to study the action of the plant pathogen effector HopM1 in Saccharomyces cerevisiae. Pseudomonas syringae, causative agent of bacterial speck in tomatoes, utilizes the type III secretion system to shuttle the effector proteins into the host cell. When expressed in yeast, HopM1 is lethal on solid media at 21°C, but not at 30°C and 37°C. The same temperature sensitive ability of HopM1 to cause death on solid media is also observed in liquid. As demonstrated by SDS PAGE-Western blot analysis, HopM1 protein is present at 21°C, 30°C and 37°C. At 21°C, a full-length protein of 78kDA is observed. At 30°C and 37°C, the majority of HopM1 protein exists as degraded fragments. HopM1 containing strains were visualized using the V5 epitope and immunofluorescent microscopy. HopM1 localizes to mitochondria and secretory organelles. This result was confirmed using cellular fractionation and sucrose gradient density centrifugation. When plated on media containg glycerol, we observed no change in expression of HopM1, thus indicating that is it unlikely that binding to mitochondria results in the lethal phenotype. We have isolated 19 spontaneous suppressor strains that are capable of surviving the HopM1 imposed lethality at 21°C. All strains have been examined for HopM1 protein expression, of which 13 express full-length HopM1 at 21°C, and 5 do not. SupM1-16, showed a significant increse in growth rates as compared to the wild type strain expressing HopM1. None of the suppressor strains show a change in localization of HopM1 as compared to wild type. One of the suppressor stains, SupM1-16 was sequenced to identify the gene(s) responsible for the suppression phenotype. Six genes that may be the suppressor gene were identified. The most likely candidate is RSP5; an E3 Ubiquitin Ligase. RSP5 contains a single mutation that changes a Glycine to Valine in the HECT domain. Overall our findings suggest that HopM1 kills the yeast cell by perturbing a secrectory pathway regulator and that mutation of RSP5 alters HopM1 effects on this pathway to allow survival.
Author: Alexandre Robert-Seilaniantz
Publisher:
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Category :
Languages : en
Pages : 234
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Author: Robert N. Goodman
Publisher: American Phytopathological Society
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Category : Medical
Languages : en
Pages : 266
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Nikolai Gogol’s short story is a sublime work of tragi-comedy. In it, he brilliantly ridicules the Ukrainian passion for litigation and reveals life as something really rather absurd. Ivan Ivanovich and Ivan Nikiforovich are the greatest of friends—until the day they begin a foolish quarrel that culminates in that very worst of insults: “And you, Ivan Ivanovich, are a goose.” From that moment on, not another word is spoken between them as they choose instead to fight out their differences in the courts. But it seems theirs is a lawsuit that is set to run for years and years.
Author: Misty D. Janes Wehling
Publisher:
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Category :
Languages : en
Pages : 328
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