Cloning and expression of Bacillus subtilis phage SPP1 in E. coli. II. Expression of lambda/SPP1 hybrid phages in E. coli minicells.

In the preceding paper (Amann et al. 1981) we described the in vitro construction of hybrids between Escherichia coli phage lambda NM607 imm434 and B. subtilis phage SPP1. These lambda/SPP1 hybrids have been used to infect minicells produced by E. coli strain DS410. Analysis on polyacrylamide gels of 35S-methionine labeled proteins synthesized in infected minicells revealed the expression of both lambda and SPP1 genes. Infection of E. coli minicells carrying plasmid pGY101, which encodes and expresses the repressor gene of phage 434, results in the selective expression of the cloned SPP1 DNA. This has resulted in the assignment of 26 out of a total of 46 known SPP1 polypeptides (Mertens et al. 1979) to individual SPP1 DNA fragments. In addition, several lambda/SPP1 fusion peptides whose transcription either originates from lambda promoters or from promoters located on the inserted SPP1 fragment, were identified.