Publication details
Cloning and expression of endolysin and its SH3 domain from polyvalent staphylococcal bacteriophage
| Authors | |
|---|---|
| Year of publication | 2010 |
| Type | Article in Proceedings |
| Conference | XIV. Setkání biochemiků a molekulárních biologů. Sborník příspěvků |
| MU Faculty or unit | |
| Citation | |
| Web | http://orion.chemi.muni.cz/Setkani/index.htm |
| Field | Genetics and molecular biology |
| Keywords | bacteriophage; Staphylococcus aureus; bacteriolysis; lytic enzymes; protein preparetion |
| Description | Endolysins are phage lytic proteins which digest the cell wall of bacteria. Here we report expression of the genes encoding lytic proteins from bacteriophage 812. This enzymes lyses cells of Staphylococcus aureus and could have practical use in phage therapy. Activity of lytic enzymes was tested on different S. aureus strains that are insensitive to wild-type phage 812 and are resistant to antibiotics. The enzymes were active against a broader range of bacterial strains than the phage itself. Lytic protein of phage 812 includes three domains. SH3-domain is probably cell wall targeting domain, which could be responsible for binding of endolysin to peptidoglycan. SH3b-domain of phage 812 has been cloned and expressed as a soluble protein in E. coli. Protein was purified to homogeneity using ammonium sulphate precipitation, anion exchange chromatography, cation exchange chromatography, and gel filtration. |
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