Abstract: The E. faecalis MSF1 bacteriophage was isolated from the raw milk and partially characterized. The bacteriophage produces small turbid plaques on the 47/3 host strain of E. faecalis. The genome of bacteriophage is formed by double stranded DNA in size between 40 to 50 kbp. Despite MSF bacteriophage is the first E. faecalis bacteriophage isolated from the milk it is highly related to E. faecalis bacteriophages and prophages isolated from other environments. MSF1 bacteriophage lytic module was characterized which is composed of family 4 holin and a muramidase. Site specific DNA methyltransferase was detected in the genome of MSF1 bacteriophage indicating that site specific DNA modification is used by bacteriophage to protect it from host encoded restriction and modification systems.