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.

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

Andrea Vannini;Biljana Petrovic;Valentina Gatta;Valerio Leoni;Simona Pepe;Laura Menotti;Gabriella Campadelli-Fiume;Tatiana Gianni
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.
Herpes Simplex Virus
153
168
METHODS IN MOLECULAR BIOLOGY
Andrea Vannini, Biljana Petrovic, Valentina Gatta, Valerio Leoni, Simona Pepe, Laura Menotti, Gabriella Campadelli-Fiume, Tatiana Gianni
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/708240
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