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Isolation of Temperature-sensitive Mutants and the Construction of a Preliminary Genetic Map for Influenza Virus

Department of Microbiology, John Curtin School of Medical Research, Australian National University, Canberra

Sixteen ts mutants were isolated from influenza virus type A₀, strain WSN, after treatment with either fluorouracil or hydroxylamine. Cells doubly infected with several pairs of ts mutants yielded a high proportion of wild-type virus, probably because of genetic recombination. Recombination frequencies with particular pairs of mutants were very variable, even in replicate tubes of the same experiment, until infected cells were treated with receptor-destroying enzyme (RDE) after adsorption of virus. Day-to-day variation between experiments could not be eliminated, but was allowed for by including a standard cross in all experiments, and relating recombination frequencies to the values obtained in this cross. Standardized recombination frequencies were then reproducible and characteristic of each pair of mutants.

The recombination frequencies thus obtained allowed certain of the mutants to be placed in a sequence with additivity between the intervals, suggesting that influenza virus may have a linear genetic map.

^* Present address: Department of Virology, The Public Health Research Institute of The City of New York, Inc. 455 First Avenue, New York, New York 10016.

Received 25 June 1969; accepted 19 August 1969.