Molecular and General Genetics MGG

, Volume 182, Issue 2, pp 299-303

First online:

Cloning and expression of Bacillus subtilis phage SPP1 in E. coli

II. Expression of λ/SPP1 hybrid phages in E. coli minicells
  • Egon P. AmannAffiliated withMax Planck Institut für Molekulare Genetik
  • , John N. ReeveAffiliated withMax Planck Institut für Molekulare Genetik

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In the preceding paper (Amann et al. 1981) we described the in vitro construction of hybrids between Escherichia coli phage λNM607 imm434 and B. subtilis phage SPP1. These λ/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 λ 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 λ/SPP1 fusion peptides whose transcription either originates from λ promoters or from promoters located on the inserted SPP1 fragment, were identified.