Molecular analyses of Chlamydiae psittaci isolates in Slovenia = Molekularnobiološki opis sevov bakterije Chlamydia psittaci, izoliranih iz papig v Sloveniji
Dovč, Alenka ; Dovč, Peter, 1958-
Seve bakterije Chlamydia psittaci (C. psittaci), izolirane iz papig, smo namnožili v laboratorijskih miših seva BALB/c in v celičnih kulturah McCoy. Po izolaciji DNK smo z metodo polimerazne verižne ...reakcije (PCR) pomnožili gen za glavni zunanji membranski protein (major outer membrane protein - MOMP) in del gena za 16S rRNK C. psittaci. Za anlizo restrikcijskih fragmenotv gena za MOMP smo uporabljali encime HaeIII, HinfI in DdeI. Med našimi sedmimi izolati nismo ugotovili vidnih razlik v vzorcu restrikcijskih fragmenotv, potrdili pa smo ujemanje z restrikcijskim vzorcem, ki je bil v literaturi opisan za izolate iz papig. Določili smo nukleotidno zaporedje 119 baznih parov (bp) dolgega fragmenta za 16S rRNK in med našimi izolati našli dve točkovni mutaciji, po primerjavi z nukleotidnimi zaporedji, deponiranimi v genski banki pa šest točkovnih mutacij. Na osnovi sekvence 96 bp dolgega fragmenta smo sestavili dendrogram, ki nakazuje podobnost med sevi C. psittaci in večje razlike do Chlamydia trachomatis (C. trachomatis), Chlamydia pneumoniae (C. pneumoniae) in Chlamydia pecorum (C. pecorum). Ta delitev je bila še nazornejša na filogenetskem drevesu, ki smo ga sestavili na osnovi celotnega gena iz 16S rRNK 15 sevov iz rodu Chlamydia. Naše delo potrjuje uporanost molekularnih metod za diagnostične namene in kaže na veliko sorodnost izolatov C. psittaci iz papig v proučevanem materialu.
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