Restriction endonuclease mapping of bacteriophage phi105 and closely related temperate Bacillus subtilis bacteriophages rho10 and rho14

Abstract

Cleavage maps of the three similar Bacillus subtilis temperate bacteriophages, 4105, plO, and p14, were constructed by partial digestion analysis utilizing the restriction endonuclease EcoRI. Comparison of the topography of these maps indicates that all phage DNAs possess cohesive ends and a number of EcoRI restriction sites; the fragments are conserved, and the estimated base substitution/nucleotide divergence between these phages is 0.03 to 0.07 based on conserved fragments or between 0.03 and 0.11 based on conserved cleavage sites. These lines of evidence indicate that 4105, plO, and p14 are closely related. Double-enzyme digestion analysis reveals that p14 DNA has unique SalGI and BglJI restriction sites and 4105 DNA has a unique SalGI restriction site, making these phages possible cloning vectors for B. subtilis.With the advent of restriction endonucleases, physical maps of various bacteriophages have been constructed: 429 (12). One use of restriction maps is in the analysis of related phage DNAs (15). Van den Hondel and Schoenmaker (24) used extensive mapping in quantitating the similarities and differences of the closely related filamentous bacteriophages M13, fd, fl, and Z J/2. More recently, Godson (5, 6) used the same technique to determine the base sequence divergence between the phages OX174, S13, and on August 4, 2020 by guest

Streptococcus plasmid pAM alpha 1 is a composite of two separable replicons, one of which is closely related to Bacillus plasmid pBC16

“…Restriction endonuclease mapping. The relative positions of HpaII cleavage sites in pAMal, pAMalA1, pAMalA2, and pAMal::Tn917 were determined from the analysis of enzyme digestions run to completion or prematurely terminated at various times, as described previously (21). BamHI, EcoRl, PvuII, PstI, HindIII, and XbaI cleavage sites were then ordered by using those enzymes individually, in combination with each other, and in combination with HpaII.…”

Correlated genetic and EcoRI cleavage map of Bacillus subtilis bacteriophage phi105 DNA

“…To further aid in visualizing G and H, the lower portion of the figure was photographically overexposed to enhance the contrast. The previously determined(16) sizes of the fragments in millions of daltons are: (A)11.0; (B) 7.1; (C) 5.8; (D) 5.2; (E) 3.5; (F) 2.1; (G) 0.6; and (H) 0.4.…”

Reorienting and expanding the physical map of temperate Bacillus subtilis bacteriophage phi 105

“…Therefore, the confirmed order of HpaI fragments is D-I-C-H-G-A-F-E-J, and the EcoRI fragment order is D-G-I-E-B-H-F-C. We have no evidence of the EcoRI-J fragment reported by Scher et al (15), and we conclude that our previous inclusion of EcoRI-J, based on partial digests, was incorrect. In retrospect, the molecular weights of several partial digestion fragments (12) would support arguments for other arrangements of the left subterminal fragments.…”

“…Physical maps of 4105. The top line is the EcoRI map of Perkins et al (12). The second line is the reoriented EcoRI map based on data in this paper.…”