1. Rescue, Purification, and Characterization of a Recombinant HSV Expressing a Transgenic Protein
- Author
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Tatiana Gianni, Laura Menotti, Valentina Gatta, Simona Pepe, Biljana Petrovic, Gabriella Campadelli-Fiume, Andrea Vannini, Valerio Leoni, Diefenbach R., Fraefel C. (eds), and Andrea Vannini, Biljana Petrovic, Valentina Gatta, Valerio Leoni, Simona Pepe, Laura Menotti, Gabriella Campadelli-Fiume, Tatiana Gianni
- Subjects
0301 basic medicine ,Virus quantification ,Transgene ,Biology ,Recombinant virus ,Genome ,Recombineering ,law.invention ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,mIL12 transgene expression ,In vivo ,law ,Plaque purification ,030220 oncology & carcinogenesis ,Recombinant DNA ,Interleukin 12 ,HSV rescue ,Virion purification ,Plaque assay - 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.
- Published
- 2019