Isolation and Characterization of Bacillus subtilis Phage from Soil Cultivated with Liquorices Root

Abstract

One virulent phage specific for Bacillus Subtilis was isolated from soil cultivated with liquorices plants in Matruh governorate, Egypt. The B. subtilis phage isolate produced 0.13 cm diameter circular plaques; turbid and without central zone. In vitro determination of thermal inactivation point (TIP), dilution end point (DEP) and longevity in vitro (LIV) were found to be 68°C/10 min, 10-4 and 96 hrs, respectively. B. subtilis phage isolate has a narrow host range and infected only the B. subtilis. Transmission electron microscopy examination showed that the purified phage is a tailed, isometric shape head and the diameter was 59 nm. In protein pattern characterization of purified B. subtilis phage isolate showed 8 structural proteins with different molecular weights when compared with protein marker. In RAPD-PCR, only eight primers succeeded to generate polymorphic DNA products. Thirty five bands were unique polymorphism with all primer used and twenty three bands were monomorphic.