DIRECT SEQUENCING OF BUTYRIVIBRIO-FIBRISOLVENS 16S RIBOSOMAL-RNA GENES AND THEIR 3'-FLANKING AND 5'-FLANKING REGIONS

Direct genomic DNA sequencing (DGS) of Butyrivibrio fibrisolvens 16S r DNA genes, and of regions 3' and 5' to the genes, was achieved using a linear amplification sequencing reaction with a single oligonucleotid e primer, without prior cloning or amplification of the target sequenc e. The utility of the method was demonstrated by comparison with PCR-a mplified template sequencing methods. Sequence differences were reveal ed by DGS between 16S rRNA genes from two strains of the rumen bacteri um Butyrivibrio fibrisolvens isolated in different countries. Using DG S we were able to determine the sequence of the 5'- and 3'-termini and flanking regions, as well as the sequence of the gene itself.