H2, a Temperate Bacteriophage Isolated from Bacillus amyloliquefaciens Strain H

Abstract

Bacillus amyloliquefaciens strain H is lysogenic for a large temperate phage we call H2. H2 has a polyhedral head 85 nm in diameter and a tail of about 17 x 434 nm. H2 lysogenizes Bacillus subtilis between the tyrA and metB genes, and gives specialized transduction of metB and, at lower frequencies, of ilvD and ilvA. The phage carries a thymidylate synthase gene and converts thymine auxotrophs of B. subtilis to prototrophy. The H2 genome is a linear DNA molecule about 129 kb in length. DNA extracted from phage particles grown in B. subtilis is not cut by the restriction endonucleases HaeIII, Fnu4HI, Bsp1286I, and BamHI; the latter enzyme is produced by B. amyloliquefaciens strain H. The prophage in lysogenic B. subtilis cells can be cut by these enzymes. We have isolated H2 mutants that carry the transposon Tn917, or a mutation resulting in clear-plaque morphology, or both.

Genome Homology and Host Range of Some SPbeta-related Bacteriophages of Bacillus Subtilis and Bacillus amyloliquefaciens

“…The group III phages have evolved several distinct means of overcoming host defences: (i) alteration of the phage adsorption proteins to permit recognition of different adsorption sites, presumably giving them the advantage of a wider host range, (ii) differential methylation of the phage DNA (G/inthert & Trautner, 1984: Noyer-Weidner et al, 1985Gfinthert et al, 1986;Weiner, 1986;Gfinthert & Reiners, 1987) allowing them to overcome host restriction enzyme barriers, and (iii) a rarity of specific restriction target sites (for example AvalI sites in H2 and SPfl ;Fink et al, 1981 ;Zahler et al, 1987b) for enzymes that may be produced by prospective hosts.…”

“…Bacillus subtilis strain CU4344 (derived from strain CU1050; Zahler et al, 1977) was used for assaying phages SPfl and SPR. Bacillus subtilis strain CU4345 (derived from strain CU3431 ; Zahler et al, 1987b) was used to assay phage H2.…”

“…We have already described derivatives of SPfl and H2 that carry the Streptococcusfaecalis transposon Tn917 (Zahler et al, , 1987b. Tn917 confers resistance to the MLS group of antibiotics.…”

“…In addition, at least some of these phages can transduce bacterial markers that are adjacent to their attachment sites in the B. subtilis chromosome (Zahler et al, 1977(Zahler et al, , 1987bRosenthal et aL, 1979;Lipsky et al, 1981;Fink & Zahler, 1982;, and in cytoplasmic plasmids (M. Weiner, data not shown). The prophages of SPfl and Z encode a bacteriocin that kills host bacteria that are non-lysogenic for these phages (Hemphill et al, 1980).…”

Temperate Bacteriophages of Bacillus amylolquefaciens

“…This choice was made partly on the basis of the ability of the parent strain H to grow well as isolated colonies on B. subtilis minimal medium (Anagnostopoulos & Spizizen, 1961), and partly because strain H is the nominal origin of the restriction endonuclease BarnHI. The results of these studies are described in the accompanying paper (Zahler et al, 1987). This work was supported by a grant from Dow Chemical Company.…”