Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020:2060:153-168.
doi: 10.1007/978-1-4939-9814-2_8.

Rescue, Purification, and Characterization of a Recombinant HSV Expressing a Transgenic Protein

Andrea Vannini  1 , Biljana Petrovic  2 , Valentina Gatta  1 , Valerio Leoni  1 , Simona Pepe  1 , Laura Menotti  3 , Gabriella Campadelli-Fiume  4 , Tatiana Gianni  1
Affiliations

Rescue, Purification, and Characterization of a Recombinant HSV Expressing a Transgenic Protein

Andrea Vannini et al. Methods Mol Biol. 2020.

Abstract

In the previous chapter, we describe the engineering of a HSV-BAC genome by galK recombineering. Here we describe the procedures to reconstitute, or regenerate, the replicating recombinant virus, and the methods to purify it and characterize it for the correct expression of the transgene. We present the example of R-115, a recombinant expressing murine interleukin 12 (mIL12) from the US1-US2 intergenic region. A specific method for the production of highly purified virions by iodixanol gradient, suitable for in vivo applications, is also detailed.

Keywords: HSV rescue; Plaque assay; Plaque purification; Virion purification; mIL12 transgene expression.

PubMed Disclaimer

Publication types

LinkOut - more resources