Cloning of the Bacillus subtilis lys and spoIIIB Genes in Phage 105

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Abstract
SUMMARY: The lys gene of Bacillus subtilis was inserted into prophage ϕ105. The recombinant phage (ϕ105dlys) 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 used to clone, using a similar technique, the spoIIIB gene(s). The second recombinant phage, ϕ105dspoIIIB, 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 ϕ105dspoIIIB transduced cells of a spoIIIB2 recipient to Spo+. In these transductants the spoIIIB2 mutation was complemented, and the cells sporulated normally.