Summary
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.
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Amann EP, Reeve JN, Morelli G, Behrens B, Trautner TA (1981) Cloning and expression of the B. subtilis phage SPP1 in E. coli. I. A company paper. Construction and characterization of λ/SPP1 hybrids. Mol Gen Genet 182:293–298
Bailone A, Levine A, Devoret R (1979) Inactivation of prophage lambda repressor in vivo. J Mol Biol 131:553–572
Behrens B, Lüder G, Behncke M, Trautner TA, Ganesan AT (1979) The genome of B. subtilis phage SPP1. Physical arrangement of phage genes. Mol Gen Genet 175:351–357
Bonner WM, Laskey RA (1974) A film detection method for tritiumlabelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem 46:83–88
Bukhari AI, Zipser D (1973) Mutants of E. coli with a defect in the degradation of nonsense fragments. Nature N Biol 243:238–241
Burger KJ, Trautner TA (1978) Specific labelling of replicating SPP1 DNA. Mol Gen Genet 166:277–285
Esche H (1975) Gene expression of bacteriophage SPP1. II. Regulatory aspects. Mol Gen Genet 142:57–66
Esche H, Schweiger M, Trautner TA (1975) Gene expression of bacteriophage SPP1. I. Phage direct protein synthesis. Mol Gen Genet 142:45–55
Ganesan AT, Anderson JJ, Luh J, Effron M (1976) DNA metabolism in Bacillus subtilis and its phage SPP1. In: Microbiology. American Society for Microbiology pp 319–325
Herrlich P, Schweiger M (1974) DNA- and RNA-directed synthesis in vitro of phage enzymes. In: Methods in Enzymology 30:654–669
Kowit JD, Goldberg AL (1977) Intermediate steps in the degradation of a specific abnormal protein in E. coli. J Biol Chem 252:8350–8351
Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227:680–685
Levine A, Bailone A, Devoret R (1979) Cellular levels of the prophage lambda and 434 repressors. J Mol Biol 131:665–661
McIntosh PK, Dunker R, Mulder C, Brown NC (1978) DNA of B. subtilis bacteriophage SPP1. Physical mapping and localization of the origin of replication. J Virol 28:865–876
Maegher RB, Tait RC, Berlach M, Boyer HW (1977) Protein expression in E. coli minicells by recombinant plasmids. Cell 10:521–536
Mertens G, Reeve JN (1977) Synthesis of cell envelope components by anucleate cells (minicells) of B. subtilis. J Bacteriol 129:1198–1207
Mertens G, Amann EP, Reeve JN (1979) Bacteriophage SPP1 polypeptides synthesized in infected minicells and in vitro. Mol Gen Genet 172:271–279
Milanesi G, Cassani G (1972) Transcription after bacteriophage SPP1 infection in B. subtilis. J Virol 10:187–192
Montenegro MA, Esche H, Trautner TA (1976) Induction of mutations in B. subtilis phage SPP1 by growth on host cells carrying a mutator DNA polymerase III. Mol Gen Genet 149:131–134
Morelli G, Montenegro MA, Hillenbrandt G, Scherzinger E, Trautner TA (1978) The genome of B. subtilis phage SPP1: Assignment of 5′-3′-polarity to the complementary strands of SPP1 DNA. Mol Gen Genet 164:93–97
Morelli G, Fisseau C, Behrens B, Trautner TA, Luh J, Ratcliff SW, Allison DP, Ganesan AT (1979) The genome of B. subtilis phage SPP1: The topology of DNA molecules. Mol Gen Genet 168:153–164
Murray NE, Brammar WJ, Murray K (1977) Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet 150:53–61
Ratcliff SW, Luh J, Ganesan AT, Behrens B, Thompson R, Montenegro MA, Morelli G, Trautner RA (1979) The genome of B. subtilis phage SPP1: The arrangement of restriction endonuclease generated fragments. Mol Gen Genet 168:165–172
Reeve, JN (1978) Selective expression of transduced or cloned DNA in minicells containing plasmid pKB280. Nature 276:728–729
Reeve JN (1977) Bacteriophage infection of minicells: A general method for identification of in vivo bacteriophage directed polypeptide biosynthesis. Mol Gen Genet 158:73–79
Reeve JN (1979) Use of minicells for bacteriophage directed polypeptide synthesis. Methods in Enzymology 68:493–503
Reeve JN, Shaw JE (1979) Lambda encodes an outer membrane protein: the lom gene. Mol Gen Genet 172:243–248
Reeve JN, Mendelson NH, Coyne SI, Hallock LL, Cole RM (1973) Minicells of B. subtilis. J Bacteriol 114:860–873
Rowley SD, Brown NC (1977) B. subtilis DNA polymerase III is required for the replication of DNA of bacteriophage SPP1 and 105. J Virol 21:493–496
Studier FW (1973) Analysis of bacteriophage T7 early mRNAs and proteins on slab gels. J Mol Biol 79:237–248
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Communicated by G. O'Donovan
Part of this work was taken from the doctoral thesis of E.A. submitted to the Freie Universität, Berlin 1979
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Amann, E.P., Reeve, J.N. Cloning and expression of Bacillus subtilis phage SPP1 in E. coli . Molec. Gen. Genet. 182, 299–303 (1981). https://doi.org/10.1007/BF00269674
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DOI: https://doi.org/10.1007/BF00269674