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

Abstract

Adenovirus type 5 DNA has low infectivity which can be increased by various techniques, one of which is the dimethyl sulfoxide (DMSO) boost. DMSO treatment of adenovirus 5 DNA-infected [human cervical carcinoma] HeLa cells results in a 10-fold increase in plaque formation, and 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 resulted in the assignment of 6 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.