Publication details
Analýza genomu a proteomu polyvalentního stafylokokového bakteriofága 812
| Title in English | Genomic and proteomic characterization of polyvalent staphylococcal bacteriophage 812 |
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| Authors | |
| Year of publication | 2005 |
| Type | Article in Periodical |
| Magazine / Source | Informační listy Česko-Slovenské genetické společnosti Gregora Mendela. |
| MU Faculty or unit | |
| Citation | |
| Field | Genetics and molecular biology |
| Keywords | bacteriophage; phage therapy; MALDI-TOF; proteomics; genome |
| Description | Virulent and polyvalent staphylophage 812, a member of Myoviridae family, exhibits strong lytic activity on a broad host range of Staphylococcus species, thus being a suitable candidate for treatment of staphylococcal infections. Genome of this phage is represented by linear 145 kb dsDNA. Our study was focused on genome and proteome comparison of 812 wild-type phage, its host-range mutants and related phages U16, 131 and SK311 with the aim to explain molecular basis of differences in their host-range. Comparison of the phage genomes revealed several rearrangements of DNA sequences as well as single nucleotide substitutions in different parts of the genomes. Nucleotide insertions and deletions of various size (~1 to 3 kb) were most frequently found in non-coding terminal genomic regions in phages with different host range. We tried to find possible correlations between mutational changes on DNA and proteome composition of the phages. After SDS-PAGE separation of phage virions digests 15 protein bands with the mass ranging from 15 to 80 kDa were identified. Twelve bands were excised for further analysis by in-gel digestion with consecutive MALDI-TOF MS. Three of the proteins designated 812_mtsp (64.4 kDa), 812_mcp (51.2 kDa) and 812_ORF8 (15.9 kDa) were identified by peptide mapping. Two of them were determined as the major capsid protein and the major tail sheath protein, respectively. Both the proteins show high similarity with the corresponding proteins of the staphylococcal polyvalent phages TWORT and K as well as those of Listeria monocytogenes phage A511. SDS-PAGE revealed size differences of the tail sheat proteins in phages 812, SK311 and U16 which substantially differ in their host range. Thus, the structural changes of tail sheat proteins should be considered as a factor influencing the host range of polyvalent staphylophages. |
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