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The Characterization of a Set of Amber Mutants of Bacteriophage T5

Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, U.K.

A collection of amber mutants of bacteriophage T5 was analysed using an in vivo complementation test and assigned to 21 complementation groups. The incomplete phage structures produced by T5 amber mutant infection of the non-permissive host were examined. The mutants were allocated to four phenotypic types, as defined by an in vitro complementation test: those which produced functional heads, H; those which produced functional tails, T; those which produced inactive heads and functional tails, HI + T; and those which did not synthesize either heads or tails, 0. Functional heads and inactive heads were indistinguishable in shape and size in the electron microscope. Four different patterns of DNA metabolism were observed when different amber mutants were grown under non-permissive conditions. They were the wild-type pattern (D⁺) in which host DNA degradation was followed by synthesis of phage DNA, host DNA degradation without phage DNA synthesis (D0), neither host DNA degradation nor phage DNA synthesis (DD0) and host DNA degradation with slight DNA synthesis (DS). Upon infection of the non-permissive host with representatives of different complementation groups of mutants either normal lysis occurred or bacterial growth ceased without subsequent lysis. The phenotypic characteristics of the amber mutants were used for partial elucidation of the functions of the affected genes.

Keywords: phage T5, amber mutants, complementation in vivo and in vitro

Received 17 December 1985; accepted 23 April 1986.