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Author: Vanessa Revindran
Publisher:
ISBN:
Category : Biochemistry
Languages : en
Pages : 262
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Book Description
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: Vanessa Revindran
Publisher:
ISBN:
Category : Biochemistry
Languages : en
Pages : 262
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Book Description
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: Vanessa Revindran
Publisher:
ISBN:
Category : Biochemistry
Languages : en
Pages : 0
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Book Description
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: J. Robin Harris
Publisher: Springer Nature
ISBN: 3030589714
Category : Science
Languages : en
Pages : 580
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Book Description
This book covers important topics such as the dynamic structure and function of the 26S proteasome, the DNA replication machine: structure and dynamic function and the structural organization and protein–protein interactions in the human adenovirus capsid, to mention but a few. The 18 chapters included here, written by experts in their specific field, are at the forefront of scientific knowledge. The impressive integration of structural data from X-ray crystallography with that from cryo-electron microscopy is apparent throughout the book. In addition, functional aspects are also given a high priority. Chapter 1 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Author: Monica A. Hughes
Publisher: Prentice Hall
ISBN:
Category : Science
Languages : en
Pages : 252
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Book Description
This text, concisely sets out the fundamentals required by students in this rapidly growing field. Plant Molecular Genetics is split into four parts: the first deals with the structure and inheritance of plant genomes; the second with the biology of Agrobacterium tumefaciens and its use in plant transformation; the third with key topics in plant molecular biology, including nitrogen fixation, the effect of light on plant development, flowering, breeding systems and disease resistance. The final section provides an overview of plant biotechnology, including a discussion of its future prospects.
Author: Andreas Bresinsky
Publisher: Springer
ISBN:
Category : Science
Languages : en
Pages : 628
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Book Description
Structure, physiology, evolution, systematics, ecology.
Author: Carl Feldherr
Publisher: Academic Press
ISBN: 0323152988
Category : Science
Languages : en
Pages : 385
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Book Description
Nuclear Trafficking is a summary of the state of knowledge in nuclear trafficking, and is organized into five parts. The book begins by discussing the diffusion and signal-mediated transport through the pores. It then looks into the detailed accounts of pore structure and composition, nuclear localization signals, signal binding proteins, RNA efflux, and biochemical factors influencing nucleocytoplasmic exchange. This book will be very useful to those people new to this field of interest.
Author: Thomas Boller
Publisher: Springer Science & Business Media
ISBN: 3709166845
Category : Science
Languages : en
Pages : 368
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Book Description
Many fungi and bacteria that associate with plants are potentially harmful and can cause disease, while others enter into mutually beneficial sym bioses. Co-evolution of plants with pathogenic and symbiotic microbes has lead to refined mechanisms of reciprocal recognition, defense and counter defense. Genes in both partners determine and regulate these mechanisms. A detailed understanding of these genes provides basic biological insights as well as a starting point for developing novel methods of crop protection against pathogens. This volume deals with defense-related genes of plants and their regulation as well as with the genes of microbes involved in their interaction with plants. Our discussion begins at the level of populations and addresses the complex interaction of plant and microbial genes in multigenic disease resistance and its significance for crop protection as compared to mono genic resistance (Chap. 1). Although monogenic disease resistance may have its problems in the practice of crop protection, it is appealing to the experimentalist: in the so-called gene-for-gene systems, single genes in the plant and in the pathogen specify the compatibility or incompatibility of an interaction providing an ideal experimental system for studying events at the molecular level (Chaps. 2 and 4). Good progress has been made in identifying viral, bacterial, and fungal genes important in virulence and host range (Chaps. 3-6). An important aspect of plant-microbe interactions is the exchange of chemical signals. Microbes can respond to chemical signals of plant origin.
Author: Paul Birch
Publisher: Humana Press
ISBN: 9781627039871
Category : Science
Languages : en
Pages : 306
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Book Description
Plant-Pathogen Interactions: Methods and Protocols, Second Edition expands upon the first edition with current, detailed protocols for the study of plant pathogen genome sequences. It contains new chapters on techniques to help identify and characterize effectors and to study their impacts on host immunity and their roles in pathogen biology. Additional chapters focus on protocols to identify avirulence and resistance genes, investigate the roles of effector targets and other defence-associated proteins in plant immunity. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Plant-Pathogen Interactions: Methods and Protocols, Second Edition seeks to aid scientists in the further study of plant immunity.
Author: Nian Wang
Publisher:
ISBN: 9780890545874
Category : Bacterial diseases of plants
Languages : en
Pages : 492
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Book Description
Author: Libo Shan
Publisher:
ISBN: 9781493968596
Category : Botany
Languages : en
Pages : 358
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Book Description
"This volume covers protocols on techniques ranging from MAMP isolations from diverse microorganisms, PRR identifications from different plant species, MAMP-PRR binding, and a series of signaling responses and events revealed by various biochemical, cellular, genetic and bioinformatic tools. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Plant Pattern Recognition Receptors: Methods and Protocolsaims to ensure successful results in the further study of this vital field." -- OCLC.
