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J Gen Virol 41 (1978), 573-586; DOI 10.1099/0022-1317-41-3-573
© 1978 Society for General Microbiology

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Mapping of Adenovirus Type 5 Temperature-sensitive Mutations by Marker Rescue in Enhanced Double DNA Infections

Janet E. Arrand

National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, U.K.

Adenovirus type 5 DNA has low infectivity (Graham & van der Eb, 1973) which can be increased by various techniques, one of which is the dimethyl sulphoxide (DMSO) boost (Stow & Wilkie, 1976). In this report, it is shown that DMSO treatment of adenovirus 5 DNA-infected HeLa cells results in a 10-fold increase in plaque formation, and that this can be used to facilitate marker rescue experiments. Double DNA infections were performed by the calcium phosphate method, co-precipitating intact temperature-sensitive mutant DNA with purified wild-type DNA restriction endonuclease fragments. Analysis of the plaquing ability of these mixtures and any progeny virus has resulted in the assignment of six temperature-sensitive mutations to discrete physical locations on the adenovirus type 5 genome. These locations are discussed with respect to the mutant phenotypes and the transcription-translation products of the appropriate regions.

Received 10 April 1978; accepted 27 June 1978.





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Copyright © 1978 by the Society for General Microbiology.