J Vet Sci.  2013 Mar;14(1):37-43. 10.4142/jvs.2013.14.1.37.

Expression and lytic efficacy assessment of the Staphylococcus aureus phage SA4 lysin gene

Affiliations
  • 1Animal Health Division, Central Institute for Research Institute on Goats, Farah 281122, India. anilmishradr@gmail.com
  • 2Division of Biological Standardization, Indian Veterinary Research Institute, Bareilly 243122, India.
  • 3Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Bareilly 243122, India.
  • 4Aquatic Animal Health and Environment Division, KRC of Central Institute of Brackish Water Aquaculture, Kakdwip 743347, India.

Abstract

Treatment of bovine mastitis caused by Staphylococcus (S.) aureus is becoming very difficult due to the emergence of multidrug-resistant strains. Hence, the search for novel therapeutic alternatives has become of great importance. Consequently, bacteriophages and their endolysins have been identified as potential therapeutic alternatives to antibiotic therapy against S. aureus. In the present study, the gene encoding lysin (LysSA4) in S. aureus phage SA4 was cloned and the nucleotide sequence was determined. Sequence analysis of the recombinant clone revealed a single 802-bp open reading frame encoding a partial protein with a calculated mass of 30 kDa. Results of this analysis also indicated that the LysSA4 sequence shared a high homology with endolysin of the GH15 phage and other reported phages. The LysSA4 gene of the SA4 phage was subsequently expressed in Escherichia coli. Recombinant LysSA4 induced the lysis of host bacteria in a spot inoculation test, indicating that the protein was expressed and functionally active. Furthermore, recombinant lysin was found to have lytic activity, albeit a low level, against mastitogenic Staphylococcus isolates of bovine origin. Data from the current study can be used to develop therapeutic tools for treating diseases caused by drug-resistant S. aureus strains.

Keyword

lysin; mastitis; phage; Staphylococcus aureus

MeSH Terms

Animals
Base Sequence
Cloning, Molecular
Gene Expression Regulation, Viral/physiology
Mucoproteins/genetics/*metabolism
Phylogeny
Polymerase Chain Reaction/methods
Recombinant Proteins
Staphylococcus Phages/genetics/*metabolism/physiology
Staphylococcus aureus/*virology
Mucoproteins
Recombinant Proteins
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