Global Analysis of Murine Cytomegalovirus Open Reading Frames Using Yeast Two-Hybrid and Growth Phenotype Analysis PDF Download
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Author: Sean Umamoto
Publisher:
ISBN:
Category :
Languages : en
Pages : 264
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Book Description
Human cytomegalovirus (HCMV), a beta-herpesvirus, is an important opportunistic pathogen that primarily affects individuals with compromised or immature immune systems. It is of great significance in AIDS patients where it can cause serious morbidity through retinitis-associated blindness, and other complications, such as pneumonia and enteritis. In developed nations, it is a leading viral cause of congenital disease, where in-utero infection manifests in mental and behavioral disorders. In order to control infection and HCMV associated disease, new compounds and novel strategies must be developed. Understanding the role viral proteins play during the course of infection will help elucidate the mechanisms of HCMV pathogenesis and provide important information on potential targets for new treatments. However, the strict species specificity of HCMV prevents any studies into the pathogenesis of the virus in an animal host. This limitation can be overcome through the use of murine cytomegalovirus (MCMV). MCMV, like HCMV, is a betaherpesvirus that exhibits similar pathogenesis in mice to HCMV infection in the human host. The genetic structure of MCMV contains significant sequence homology to HCMV AD169 in at least 78 ORFs and can thereby be used as an important tool in elucidating the functions of these ORFs in a complete in vivo system. In our study, we have conducted a comprehensive YTH screen to identify potential interactions between approximately 170 MCMV ORFs. Growth phenotype analysis were also conducted using five different cell lines potentially involved in various aspects of CMV infection. Between these 170 predicted proteins we have identified 94 potential interactions that exhibit varying levels of essentiality depending on the type of cell infected. We aim to understand the nature of the interactions between the viral particle and proteins encoded by the virus in order to elucidate potential mechanisms by which these proteins help to assemble and create new progeny viruses. The interactions that we have identified in this study provide a framework to predict the functions of uncharacterized viral proteins. And understanding the importance of each protein in the context of infection can further help to determine the nature of these unknown viral proteins. Together using information about known viral proteins that interact with these unknown elements, we can develop a better understanding of how all of these components contribute to viral infection which can be used to determine more effective methods to treat or prevent CMV associated diseases.
Author: Sean Umamoto
Publisher:
ISBN:
Category :
Languages : en
Pages : 264
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Book Description
Human cytomegalovirus (HCMV), a beta-herpesvirus, is an important opportunistic pathogen that primarily affects individuals with compromised or immature immune systems. It is of great significance in AIDS patients where it can cause serious morbidity through retinitis-associated blindness, and other complications, such as pneumonia and enteritis. In developed nations, it is a leading viral cause of congenital disease, where in-utero infection manifests in mental and behavioral disorders. In order to control infection and HCMV associated disease, new compounds and novel strategies must be developed. Understanding the role viral proteins play during the course of infection will help elucidate the mechanisms of HCMV pathogenesis and provide important information on potential targets for new treatments. However, the strict species specificity of HCMV prevents any studies into the pathogenesis of the virus in an animal host. This limitation can be overcome through the use of murine cytomegalovirus (MCMV). MCMV, like HCMV, is a betaherpesvirus that exhibits similar pathogenesis in mice to HCMV infection in the human host. The genetic structure of MCMV contains significant sequence homology to HCMV AD169 in at least 78 ORFs and can thereby be used as an important tool in elucidating the functions of these ORFs in a complete in vivo system. In our study, we have conducted a comprehensive YTH screen to identify potential interactions between approximately 170 MCMV ORFs. Growth phenotype analysis were also conducted using five different cell lines potentially involved in various aspects of CMV infection. Between these 170 predicted proteins we have identified 94 potential interactions that exhibit varying levels of essentiality depending on the type of cell infected. We aim to understand the nature of the interactions between the viral particle and proteins encoded by the virus in order to elucidate potential mechanisms by which these proteins help to assemble and create new progeny viruses. The interactions that we have identified in this study provide a framework to predict the functions of uncharacterized viral proteins. And understanding the importance of each protein in the context of infection can further help to determine the nature of these unknown viral proteins. Together using information about known viral proteins that interact with these unknown elements, we can develop a better understanding of how all of these components contribute to viral infection which can be used to determine more effective methods to treat or prevent CMV associated diseases.
Author:
Publisher:
ISBN:
Category :
Languages : en
Pages :
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Book Description
Murine cytomegalovirus (MCMV) in its natural host, the mouse, is an excellent model for studying the biology of cytomegalovirus infection. Mostly this model has been used to study gene homologues of human cytomegalovirus (HCMV). Of the predicted 170 MCMV open reading frames (ORFs) only 78 have significant amino acid identity with genes in HCMV. To better understand the biological mechanisms underlying the differences between the viruses, for example their species specificity and immune evasion genes, MCMV unique ORFs need to be examined. Here the role of m29 and m29.1 ORFs in the MCMV (Smith strain), which have no homology with ORFs of any other cytomegalovirus, have been examined. The m29 and m29.1 ORFs are overlapping and encoded on opposite strands of the double-stranded DNA genome. Sequence analysis over this region showed a discrepancy to the published sequence. An additional G (guanine) nucleotide was found at nucleotide position 36,198 that alters the predicted ORFs, m29 being 242 amino acids shorter and m29.1 210 amino acids longer than the predicted sequence. This was confirmed by sequencing the MCMV Birmingham K181 strain, the Birmingham Smith strain and MCMV wild type isolates- N1, K17A and G4. Transcripts from the newly identified m29 and m29.1 ORFs were confirmed by reverse transcriptase PCR (RT-PCR). They were produced at early (3h) and immediate-early (2h) times post-infection respectively as determined by cycloheximide and phosphonoacetic acid treatment but were continuously expressed up to at least 24h post-infection. 5' and 3'-RACE (rapid amplification of cDNA ends) analysis from m29.1 ORF confirmed the production of a ~2.4 kb transcript and a low abundance spliced transcript from which a 123bp intron had been removed. Mutants of ORF m29 and m29.1 have been produced in which ET recombination was used to introduce stop codon mutations within these overlapping ORFs. This was achieved by single base alterations near to the 5` end of each ORF that prev.
Author: Ann Arvin
Publisher: Cambridge University Press
ISBN: 1139461648
Category : Medical
Languages : en
Pages : 1325
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Book Description
This comprehensive account of the human herpesviruses provides an encyclopedic overview of their basic virology and clinical manifestations. This group of viruses includes human simplex type 1 and 2, Epstein–Barr virus, Kaposi's Sarcoma-associated herpesvirus, cytomegalovirus, HHV6A, 6B and 7, and varicella-zoster virus. The viral diseases and cancers they cause are significant and often recurrent. Their prevalence in the developed world accounts for a major burden of disease, and as a result there is a great deal of research into the pathophysiology of infection and immunobiology. Another important area covered within this volume concerns antiviral therapy and the development of vaccines. All these aspects are covered in depth, both scientifically and in terms of clinical guidelines for patient care. The text is illustrated generously throughout and is fully referenced to the latest research and developments.
Author: National Research Council
Publisher: National Academies Press
ISBN: 0309070864
Category : Nature
Languages : en
Pages : 348
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Book Description
Scientific Frontiers in Developmental Toxicology and Risk Assessment reviews advances made during the last 10-15 years in fields such as developmental biology, molecular biology, and genetics. It describes a novel approach for how these advances might be used in combination with existing methodologies to further the understanding of mechanisms of developmental toxicity, to improve the assessment of chemicals for their ability to cause developmental toxicity, and to improve risk assessment for developmental defects. For example, based on the recent advances, even the smallest, simplest laboratory animals such as the fruit fly, roundworm, and zebrafish might be able to serve as developmental toxicological models for human biological systems. Use of such organisms might allow for rapid and inexpensive testing of large numbers of chemicals for their potential to cause developmental toxicity; presently, there are little or no developmental toxicity data available for the majority of natural and manufactured chemicals in use. This new approach to developmental toxicology and risk assessment will require simultaneous research on several fronts by experts from multiple scientific disciplines, including developmental toxicologists, developmental biologists, geneticists, epidemiologists, and biostatisticians.
Author: Shmuel Razin
Publisher: Springer Science & Business Media
ISBN: 0306476061
Category : Science
Languages : en
Pages : 574
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Book Description
was the result of the efforts of Robert Cleverdon. The rapidly developing discipline of molecular biology and the rapidly expanding knowledge of the PPLO were brought together at this meeting. In addition to the PPLO specialists, the conference invited Julius Marmur to compare PPLO DNA to DNA of other organisms; David Garfinkel, who was one of the first to develop computer models of metabolism; Cyrus Levinthal to talk about coding; and Henry Quastler to discuss information theory constraints on very small cells. The conference was an announcement of the role of PPLO in the fundamental understanding of molecular biology. Looking back 40-some years to the Connecticut meeting, it was a rather bold enterprise. The meeting was international and inter-disciplinary and began a series of important collaborations with influences resonating down to the present. If I may be allowed a personal remark, it was where I first met Shmuel Razin, who has been a leading figure in the emerging mycoplasma research and a good friend. This present volume is in some ways the fulfillment of the promise of that early meeting. It is an example of the collaborative work of scientists in building an understanding of fundamental aspects of biology.
Author: James M Cregg
Publisher: Springer Science & Business Media
ISBN: 1588294293
Category : Science
Languages : en
Pages : 553
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Book Description
This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by the methods of restriction enzyme-mediated integration (REMI). Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. This volume fully updates and expands upon the first edition.
Author: Thomas E. Shenk
Publisher: Springer Science & Business Media
ISBN: 3540773495
Category : Medical
Languages : en
Pages : 477
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Book Description
This volume has gathered some of the experts in the field to review aspects of our understanding of CMV and to offer perspectives of the current problems associated with CMV. The editors and authors hope that the chapters will lead to a better understanding of the virus that will assist in the development of new and unique antivirals, a protective vaccine, and a full understanding of CMV's involvement in human disease.
Author: Bernard Roizman
Publisher: Raven Press (ID)
ISBN:
Category : Medical
Languages : en
Pages : 458
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Book Description
Author: Institute of Medicine
Publisher: National Academies Press
ISBN: 0309219396
Category : Science
Languages : en
Pages : 570
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Book Description
Many potential applications of synthetic and systems biology are relevant to the challenges associated with the detection, surveillance, and responses to emerging and re-emerging infectious diseases. On March 14 and 15, 2011, the Institute of Medicine's (IOM's) Forum on Microbial Threats convened a public workshop in Washington, DC, to explore the current state of the science of synthetic biology, including its dependency on systems biology; discussed the different approaches that scientists are taking to engineer, or reengineer, biological systems; and discussed how the tools and approaches of synthetic and systems biology were being applied to mitigate the risks associated with emerging infectious diseases. The Science and Applications of Synthetic and Systems Biology is organized into sections as a topic-by-topic distillation of the presentations and discussions that took place at the workshop. Its purpose is to present information from relevant experience, to delineate a range of pivotal issues and their respective challenges, and to offer differing perspectives on the topic as discussed and described by the workshop participants. This report also includes a collection of individually authored papers and commentary.
Author: Matthew J. Gray
Publisher: Springer
ISBN: 9783319137568
Category : Science
Languages : en
Pages : 246
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Book Description
This is the first book on ranaviruses. Ranaviruses are double-stranded DNA viruses that cause hemorrhagic disease in amphibians, reptiles, and fish. They have caused mass die-offs of ectothermic vertebrates in wild and captive populations around the globe. There is evidence that this pathogen is emerging and responsible for population declines in certain locations. Considering that amphibians and freshwater turtles are suitable hosts and the most imperiled vertebrate taxa in the world, ranaviruses can have significant impacts on biodiversity and ecosystem function. Additionally, many fish that are raised in aquaculture facilities and traded internationally are suitable hosts; thus, the potential economic impact of ranaviruses is significant. Ranaviruses also serve as a model for replication and gene function of large double-stranded DNA viruses. There is an urgent need to assemble the contemporary information on ranaviruses and provide guidance on how to assess their threats in populations. Through the Global Ranavirus Consortium, 24 experts from six countries were organize to write this volume, the first book on ranaviruses. The book begins with a discussion on the global extent of ranaviruses, case histories of infection and disease in ectothermic vertebrates, and current phylogeny. Basic principles of ranavirus ecology and evolution are covered next, with a focus on host-pathogen interactions and how the virus emerges in its environment. There are two chapters that will discuss the molecular biology of ranaviruses, host response to infection, and the genes responsible for immune system evasion. One chapter establishes standards for testing for infection and diagnosing ranaviral disease. The book ends by providing guidance on how to design ranavirus surveillance studies and analyze data to determine risk, and discussing the role of the Global Ranavirus Consortium in organizing research and outreach activities.
