Isolation, characterization and genome sequencing of phage MZTP02 from Bacillus thuringiensis MZ1

Abstract

A lysogenic phage, MZTP02, was produced via induction by mitomycin C from Bacillus thuringiensis (B. thuringiensis) strain MZ1. Plaques were about 3 mm in diameter with a small inner zone consisting of new B. thuringiensis colonies. Electron microscopic analysis showed that MZTP02 had a long tail (220 nm x 18 nm) and an icosahedral head (82 nm x 85 nm). MZTP02 was insensitive to organic solvents such as chloroform, and infected six B. thuringiensis strains. Its complete genome contained 15,717 base pairs (bp) with 37.55% G + C content. Two inverted terminal repeats consisting of 40 bp were 65% identical. Twenty putative open reading frames (ORFs) were found in the MZTP02 genome, and nine predicted proteins, including two terminase subunits, portal protein, minor head protein, scaffold protein, two putative membrane proteins, tail component, and minor structural protein, showed similarity to other phage proteins. But six ORFs were unique. The presence of a terminal protein at the 5'-terminus was demonstrated using proteinase K, lambda exonuclease and E. coli exonuclease III to digest the genome DNA. A TMP phylogenetic tree was constructed based on amino acid sequences from ten phages.