CONSTRUCTION AND PHYSICAL MAP OF A BACILLUS SUBTILIS SPECIALIZED TRANSDUCING PHAGE ρ11 CONTAINING B. SUBTILIS LYS⁺ GENE

Abstract

The lys⁺ gene was detected as the smallest DNA. fragment (2.5×10⁶ dalton) among the several genetic markers tested from EcoRI-cleaved B. subtilis chromosomal DNA preparations by agarose gel electrophoresis followed by identification with DNA mediated transformation experiments. The DNA fragment retaining the lys⁺ gene was cloned in a B. subtilis temperate phage ρ11 using the prophage transformation method. The molecular weight of the genome of the constructed specialized transducing phage (ρ11dlys⁺) was estimated to be 72-73×10⁶. The phage was cross-reacted with an antiserum against ρ11 phage particles. The physical map of ρ11dlys⁺ DNA using SalI and BamHI was quite different from that of ρ11 DNA. ρ11dlys⁺ did not carry theρ11-coded thymidyrate synthetase gene.