Publication details
Metoda přímé detekce specifických genů v koloniích bakterií
| Title in English | Method of direct detection of specific genes in bacterial colonies |
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| Authors | |
| Year of publication | 2003 |
| Type | Article in Proceedings |
| Conference | Tomáškovy dny 2003 |
| MU Faculty or unit | |
| Citation | |
| Field | Genetics and molecular biology |
| Keywords | Staphylococcus aureus; S epidermidis; S. hyicus; S intermedius; toxins |
| Description | Among a wide spectrum of molecular biology methods, the macrorestriction analysis using PFGE, and PCR amplification of specific genes encoding virulence factors are the most methods employed for the diagnostics of clinically significant bacterial genera. The key role play the usage of molecular probes and of the hybridization. In our department, the method of colony hybridization has been employed and verified, namely in toxinogenic strains of Staphylococcus spp. Hybridization experiments have been performed with digoxigenin-labeled molecular probes, derived from sequences of enterotoxin C (SEC) and thermostable nuclease (TN) genes. They allowed us to confirm the presence of sec1 and nuc genes directly in the genomes of eleven SEC- and thirteen TN-positive S. aureus strains. In two SEC-negative S. aureus strains and in the other staphylococci, i.e. S. epidermidis, S. hyicus and S. intermedius, no hybridization signals have been observed. It could be concluded, that this method gives the primary information about the presence of certain sequence in the whole genome. It is appropriate for the basic genetic screening of microorganism, especially in mixed cultures. The strength of this method consists in the saving of time and material, needful for the DNA isolation and the Southern blotting. Colony hybridization is sufficient in all the cases if no demand on the precise localisation of the sequence in the genome is required. |
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