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.
Andrea Vannini, B.P. (2020). Rescue, Purification, and Characterization of a Recombinant HSV Expressing a Transgenic Protein. New York, NY : Humana Press [10.1007/978-1-4939-9814-2_8].
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.| File | Dimensione | Formato | |
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467481_2_En_8_Chapter_Author iris.pdf
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