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丁香实验推荐阅读
Interactions of Legionella Effector Proteins with Host Phosphoinositide Lipids

By means of the Icm/Dot type IV secretion system Legionella pneumophila translocates several effector proteins into host cells, where they anchor to the cytoplasmic face of the LCV membrane by binding to phosphoinositide (PI) lipids. Thus, phosphatidylinositol-4-phosphate anch ...

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Methods to Study Legionella Transcriptome In Vitro and In Vivo

The study of transcriptome responses can provide insight into the regulatory pathways and genetic factors that contribute to a specific phenotype. For bacterial pathogens, it can identify putative new virulence systems and shed light on the mechanisms underlying the regulation of vi ...

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Co-immunoprecipitation: ProteinRNA and ProteinDNA Interaction

Transcriptional and posttranscriptional regulators play a critical role in allowing a bacterium to adapt to the diverse environments and conditions it encounters. In order to characterize the role of these regulators the identification of their specific interaction partners is ...

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Identification of Legionella Effectors Using Bioinformatic Approaches

Legionella pneumophila the causative agent of Legionnaires’ disease, actively manipulates host cell processes to establish a replication niche inside host cells. The establishment of its replication niche requires a functional Icm/Dot type IV secretion system which transloc ...

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Measurements of Vaccinia Virus Dissemination Using Whole Body Imaging: Approaches for Predicting of Lethality in Challenge Models and Testing of Vacci

Preclinical evaluation of novel anti-smallpox vaccines and antiviral treatments often rely on mouse �challenge models using pathogenic vaccinia virus, such as Western Reserve (WR) strain or other orthopoxviruses. Traditionally, efficacy of treatment is evaluated using var ...

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Mousepox, A Small Animal Model of Smallpox

Ectromelia virus infections in the laboratory mouse have emerged as a valuable model to investigate human orthopoxvirus infections to understand the progression of disease, to discover and characterize antiviral treatments, and to study the host–pathogen relationship as it rel ...

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Analyzing CD8 T Cells in Mouse Models of Poxvirus Infection

Mouse models of immunology are frequently used to study host responses to poxviruses or poxvirus-based recombinant vaccines. In this context, the magnitude of CD8+ T cell responses is often of interest. Methods to evaluate CD8+ T cell responses extend from those that rely on indirect measurem ...

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Generation and Characterization of Monoclonal Antibodies Specific for Vaccinia Virus

Monoclonal antibodies to specific vaccinia virus (VACV) proteins are valuable reagents in studies of VACV. In this chapter, we describe methods of generating a panel of monoclonal antibodies that recognize a variety of VACV proteins in their native conformation in infected cells. The ant ...

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Bioinformatics for Analysis of Poxvirus Genomes

In recent years, there have been numerous unprecedented technological advances in the field of molecular biology; these include DNA sequencing, mass spectrometry of proteins, and microarray analysis of mRNA transcripts. Perhaps, however, it is the area of genomics, which has now genera ...

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Antigen Presentation Assays to Investigate Uncharacterized Immunoregulatory Genes

Antigen presentation to T lymphocytes is the seminal triggering event of the specific immune response, and poxviruses encode immunomodulatory genes that disrupt this process. Discovery of viral proteins that interfere with steps in the antigen presentation process requires a rob ...

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Characterization of Poxvirus-Encoded Proteins that Regulate Innate Immune Signaling Pathways

Innate immune recognition of pathogens is critical to the prompt control of infections, permitting the host to survive to develop long-term immunity via an adaptive immune response. Poxviruses encode a family of proteins that inhibit signaling by Toll-like receptors to their downstre ...

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Application of Quartz Crystal Microbalance with Dissipation Monitoring Technology for Studying Interactions of Poxviral Proteins with Their Ligands

Poxviruses are one of the most complex of animal viruses and encode for over 150 proteins. The interactions of many of the poxviral-encoded proteins with host proteins, as well as with other proteins, such as transcription complexes, have been well characterized at the qualitative level. Some h ...

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Central Nervous System Distribution of the Poxviral Proteins After Intranasal Administration of Proteins and Titering of Vaccinia Virus in the Brain A

Poxviral proteins are known to interact with the immune system of the host. Some of them interact with the transcription factors of the host, whereas others interact with the components of the immune system. Vaccinia virus secretes a 28.8-kDa complement control protein (VCP), which is known to re ...

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Working Safely with Vaccinia Virus: Laboratory Technique and Review of Published Cases of Accidental Laboratory Infections

Vaccinia virus (VACV), the prototype orthopoxvirus, is widely used in the laboratory as a model system to study various aspects of viral biology and virus–host interactions, as a protein expression system, as a vaccine vector, and as an oncolytic agent. The ubiquitous use of VACVs in the laboratory ...

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In-Fusion Cloning with Vaccinia Virus DNA Polymerase

Vaccinia virus DNA polymerase (VVpol) encodes a 3′-to-5′ proofreading exonuclease that can degrade the ends of duplex DNA and expose single-stranded DNA tails. The reaction plays a critical role in promoting virus recombination in vivo because single-strand annealing reactions can th ...

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Genetic Manipulation of Poxviruses Using Bacterial Artificial Chromosome Recombineering

Traditional methods for genetic manipulation of poxviruses rely on low-frequency natural recombination in virus-infected cells. Although these powerful systems represent the technical foundation of current knowledge and applications of poxviruses, they require long (≥ ...

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Easy and Efficient Protocols for Working with Recombinant Vaccinia Virus MVA

Modified vaccinia virus Ankara (MVA) is a highly attenuated and replication-deficient strain of vaccinia virus that is increasingly used as vector for expression of recombinant genes in the research laboratory and in biomedicine for vaccine development. Major benefits of MVA inclu ...

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Screening for Vaccinia Virus Egress Inhibitors: Separation of IMV, IEV, and EEV

Concerns about the possible use of variola virus as a biological weapon as well as the need for therapeutics for the treatment or prevention of naturally acquired poxvirus infections or vaccination complications have led to the search for small molecule inhibitors of poxvirus replicati ...

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Isolation of Recombinant MVA Using F13L Selection

Modified vaccinia virus Ankara (MVA) has become a widely used vector for vaccine and laboratory purposes. Despite significant advances in recombinant MVA technology, the isolation of recombinant viruses remains a tedious and difficult process. This chapter describes the use of an eff ...

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Imaging of Vaccinia Virus Entry into HeLa Cells

The recently developed technique of live cell imaging has found numerous applications, including the detection of virus movements in living cells. To monitor virus motility, viruses or cellular proteins are fused with fluorescence markers and then detected by time-lapse fluoresce ...

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