Replication of recombinant phi 29 DNA molecules in Bacillus subtilis protoplasts.

Recombinant phi 29 DNA molecules of different sizes and containing terminal protein at one or both ends, or without terminal protein, were prepared and their replication in Bacillus subtilis protoplasts was studied. Only phi 29 DNA molecules containing terminal protein at both ends replicate in vivo. The replication of symmetric DNA recombinant molecules (dimers) gives rise to displaced strands which by self-annealing create monomers with the two DNA strands covalently linked. Viral proteins p2, p3, and p6 are essential for replication of phi 29 DNA molecules in this system. Protein p17 is not essential, but stimulates the efficiency of replication. This stimulation depends on the host used.