Molecular cloning of the rRNA genes of Prevotella bryantii B14

Abstract

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.