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Methods Mol Biol. 2020;2060:219-239. doi: 10.1007/978-1-4939-9814-2_12.

HSV Mutant Generation and Dual Detection Methods for Gaining Insight into Latent/Lytic Cycles In Vivo.

Author information

1
Division of Infectious Diseases, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA. nancy.sawtell@cchmc.org.
2
Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.

Abstract

Two important components of a useful strategy to examine viral gene function, regulation, and pathogenesis in vivo are (1) a highly efficient protocol to generate viral mutants that limits undesired mutation and retains full replication competency in vivo, and (2) an efficient system to detect and quantify 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.

KEYWORDS:

E. coli β-galactosidase; Herpes simplex virus; Histochemistry; Immunohistochemistry; Latency; Local host cellular response; Lytic infection; Marker rescue; Plaque purification; Promoter/reporter; Replication kinetics; Selection marker; Sensory neuron; Tissue culture; Transfection; Trigeminal ganglion; Viral DNA; Viral gene expression; Viral mutant; Whole tissue

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