Abstract
As an inanimate virus, herpes simplex virus type 1 (HSV-1) necessarily encodes all of its functions in its DNA. Isolation of pure viral DNA allows multiple downstream applications, including the creation of recombinant HSV strains, cloning of selected regions, and sequencing of viral DNA. The term nucleocapsid refers to the combination of the viral genome with the enclosing capsid; these viral genomes are necessarily linear and have been packaged for egress, even if they are not yet released from the cell. In contrast, viral DNA that is not associated with capsids may include episomal or concatenated forms and may have modifications such as histones that are added within cells. During this protocol, the viral capsid protects the HSV genome from reagents that strip away and destroy most cellular contaminants. This procedure describes the isolation of viral nucleocapsids and their subsequent dissolution to purify clean, linear HSV DNA.
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Acknowledgement
Yolanda Tafuri helped to fine-tune the use of this protocol across many strains of HSV-1. Present and former members of Lynn Enquist’s lab at Princeton University informed this protocol, which is based on earlier work by Greg Smith and others (see References).
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Szpara, M. (2014). Isolation of Herpes Simplex Virus Nucleocapsid DNA. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus. Methods in Molecular Biology, vol 1144. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0428-0_3
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DOI: https://doi.org/10.1007/978-1-4939-0428-0_3
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