Molecular cloning of the rRNA genes of Prevotella bryantii B14 | Molekulsko kloniranje genov za rRNK bakterije Prevotella bryantii B14
2000
PETERKA, Matjaž | AVGUŠTIN, Gorazd
Inglés. Parts of the rRNA operons of anaerobic rumen bacteria Prevotella bryantii B14 were cloned. 350 colonies carrying pBluescriptII with HindIII DNA fragments were screened with 16S rDNA specific probe and six positive clones were obtained. Inserts with 16S rRNA genes were identified by hybridisation with digoxigenin labelled Prevotella-Bacteroides 16S rRNA specific probe. All six inserts were approximately 2 kb long and contained large parts of the 16S rRNA gene, 16S-23S rRNA internal spacer region and approximately 100 bp of 23S rRNA gene. The origin of 16S rRNA genes were confirmed by PCR with P. bryantii 16S rRNA specific primer.
Mostrar más [+] Menos [-]Esloveno. Klonirali smo dele operonov rRNK anaerobne vampne bakterije Prevotella bryantii B14. S sondo, specifično za gen za 16S rRNK, smo pregledali 350 bakterijskih kolonij z vektorjem pBluescripII. Našli smo šest pozitivnih klonov, katerih vektorji so vsebovali gen za 16S rRNK. Fragmente, ki so vsebovali gene za 16S rRNK, smo identificirali s hibridizacijo z digoksigeninom označeno sondo, ki je bila specifična za gen za 16S rRNK rodov Prevotella in Bacteroides. Fragmenti so bili dolgi 2 kb in so vsebovali gen za 16S rRNK, 16S-23S rRNK interno medgensko regijo in približno 100 bp gena za 23S rRNK. Poreklo genov za 16S rRNK smo potrdili z verižno reakcijo s polimerazo (PCR) z začetnim oligonukleotidom specifičnim za gen za 16S rRNK vrste P. bryantii.
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