In vitro packaging of foreign DNA into heads of bacteriophage T1.

Abstract

The isolation of a collection of 44 morphologically T1-like phages is described. It is shown that these phages share some similarity with T1 in terms of cross-inactivation with anti-T1 serum, particle proteins and DNA packaging in vitro by the headful process. Virion DNA extracted from these phages was treated with T1 in vitro packaging extracts and the reaction mixtures were tested for the formation of infectious phage particles. The packaging efficiencies observed varied from about 1 to 100% of that of virion T1 DNA. Phage lambda virion DNA was packaged with an efficiency of between 0.01 and 2% (5 X 10(1) to 3 X 10(3) p.f.u./micrograms DNA), the shorter deleted derivative lambda L47 being packaged more efficiently than normal length lambda C1857 DNA. Virion DNA from phages T3 and T7 was also packaged at an efficiency similar to that for lambda. The in vitro packaging of T1 DNA requires the presence of the pac sequence which initiates headful packaging from a concatemeric precursor. The high efficiency of packaging DNA from some of the T1-like phages may indicate the presence of similar packaging sequences. However, in the case of lambda L47, which is known not to contain such a sequence, the in vitro DNA packaging reaction must occur by a secondary pathway unrelated to the headful mechanism.