Cloning of the Bacillus subtilis lys and spoIIIB genes in phage phi 105

Abstract

The lys gene of Bacillus subtilis was inserted into prophage phi 105. The recombinant phage (phi 105dlys) contained DNA which was about 2 MDal smaller than the wild-type phage DNA, and the phage particles had no tails. The phage did not plaque but, when provided with tails in vitro, it transduced both lys-1 and lys-3 strains of B. subtilis to Lys$. The lys$ gene was located on a 2.5 MDal EcoRI restriction fragment. Subsequently this phage was phi 105 105dspoIIIB, was also defective, i.e. without tails. The DNA was 1.5 MDal smaller than the wild-type phage DNA and the spoIIIB2$ gene was located on a 3 MDal EcoRI fragment. When provided with tails in vitro, phage phi 105dspoIIIB transduced cells of a spoIIIB2 recipient to Spo$. In these transductants the spoIIIB2 mutation was complemented, and the cells sporulated normally.