Titlebook: Herpes Simplex Virus; Methods and Protocol Russell J. Diefenbach,Cornel Fraefel Book 2020Latest edition Springer Science+Business Media, LL

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Herpes Simplex Virus978-1-4939-9814-2Series ISSN 1064-3745 Series E-ISSN 1940-6029

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Methods in Molecular Biologyhttp://image.papertrans.cn/h/image/425949.jpg

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https://doi.org/10.1007/978-1-4939-9814-2Nucleocapsid DNA; Bacterial Artificial Chromosomes (BACs); Latent/Lytic Cycles In Vivo; Cryo-EM; CRISPR-

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HSV Mutant Generation and Dual Detection Methods for Gaining Insight into Latent/Lytic Cycles In Viuantify viral promoter activity and gene expression in rare cells in vivo and to gain insight into the surrounding tissue environment. Our strategy and protocols for generating, characterizing, and employing HSV viral promoter/reporter mutants in vivo are provided in this two-part chapter.

Immunization

Characterization of Extracellular HSV-1 Virions by Proteomics,of differential and density centrifugation steps. In addition, L-particles, which are viral particles devoid of a genome and capsid but present in the extracellular milieu, are depleted on Ficoll 400 gradients. As previously reported, the resulting viral particles are free of most contaminants and suitable for mass spectrometry.

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oHSV Genome Editing by Means of , Recombineering,re we describe the engineering of the HSV recombinant BAC 115 by the insertion of a heterologous cassette for the expression of murine interleukin 12 (mIL12) in the intergenic sequence between US1 and US2 ORFs.