X-ray inactivation, Weigle reactivation, and Weigle mutagenesis of the lysogenic Vibrio κ phage

Abstract

Vibrio cholerae lysogenic κ phage was inactivated by X-ray (60 kV) in a dose-dependent manner, the inactivation dose leading to 37% survival (D37) in phosphate-buffered saline (PBS), pH 7.4, being 0.36 kGy. The phages were significantly protected against X-ray irradiation when histidine or cysteine or both were present in PBS or when phages were irradiated in nutrient broth. Maximum protection was offered when both histidine (10.0 mM) and cysteine (10.0 mM) were present in PBS (dose enhancement factor being 4.17). The X-irradiated κ phages also underwent a small but significant Weigle reactivation and also Weigle mutagenesis in the UV-irradiated V. cholerae host H218Smr. The Weigle factor or the frequency of clear-plaque mutants increased with increasing UV dose, attained a maximum at a UV dose of 2.4 J m−2, and thereafter decreased gradually with a further increase of the UV dose. The X-ray dose (D) – survival (S) curves could be empirically described by the equation S = exp[−(aD + bD2)], where a and b are constants depending on the irradiation conditions, and a good agreement between the theoretical curves and experimental data was obtained. Key words: Vibrio cholerae, lysogenic κ phage, X-ray inactivation, Weigle reactivation, Weigle mutagenesis.