An efficient selection system for packaging herpes simplex virus amplicons
Zhang, Xiaoliu and O′Shea, Helen and Entwisle, Clare and Boursnell, Mike and Efstathiou, Stacey and Inglis, Stephen,, 79, 125-131 (1998),
doi = https://doi.org/10.1099/0022-1317-79-1-125,
publicationName = Microbiology Society,
issn = 0022-1317,
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 HSVthymidine 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 × 109 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.,
language=,
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