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An efficient selection system for packaging herpes simplex virus amplicons.

X Zhang·H O'Shea·C Entwisle·M Boursnell·S Efstathiou·S Inglis
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Abstract

Due to their simplicity and flexibility of genomic construction, herpes simplex virus (HSV) amplicon-based vectors are attractive vehicles for gene delivery. However, a significant problem faced in the generation of amplicon stocks is the low amplicon to helper virus (A/H) ratio. In order to improve the proportion of amplicons generated, a selection system for amplicon production was developed in which the HSV thymidine kinase (TK) gene is inserted into an amplicon plasmid and an HSV mutant with both TK and glycoprotein H (gH) genes deleted is used as a helper virus. Using a protocol in which amplicon stocks are passaged 2-3 times in BHK cells of TK- and gH+ genotype in the presence of selection medium containing methotrexate, stock preparations with high A/H ratio (up to 5:1) and high amplicon titre (>1 x 10(9) infectious units/ml) were generated. In vitro characterization demonstrated that a high level of biologically functional products can be efficiently produced from these amplicon constructs.

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AnimalsCell LineChlorocebus aethiopsCricetinaeCulture MediaGene ExpressionGenetic VectorsHelper VirusesHerpesvirus 1, HumanHumansInterleukin-2MutagenesisPlasmidsThymidine KinaseVero CellsViral Envelope ProteinsJournal Article

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X Zhang, H O'Shea, C Entwisle, M Boursnell, S Efstathiou, S Inglis. 1998. An efficient selection system for packaging herpes simplex virus amplicons.. https://doi.org/10.1099/0022-1317-79-1-125

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