title: Expression and lytic efficacy assessment of the Staphylococcus aureus phage SA4 lysin gene.
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 artial 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.
authors: Animal Health Division, Central Institute for Research Institute on Goats, Farah 281122, India. anilmishradr@gmail.com
authors: Anil Kumar
authors: AK
authors: Mishra
authors: Mayank
authors: M
authors: Rawat
authors: Konasagara Nagaleekar
authors: KN
authors: Viswas
authors: Abhishek
authors: Sujeet
authors: S
authors: Kumar
authors: Manjunatha
authors: M
authors: Reddy
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D009088:Mucoproteins / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins; D013204:Staphylococcus Phages / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D013211:Staphylococcus aureus / Q000821:virology*
chemical_list: D009088:Mucoproteins; D011994:Recombinant Proteins; C010393:lysin, gastropoda
keywords: Staphylococcus aureus; lysin; mastitis; phage
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