Direct Sequencing of Butyrivibrio fibrisolvens 16S rRNA Genes and their 3'- and 5'-Flanking Regions

Abstract

Direct genomic DNA sequencing (DGS) of Butyrivibrio fibrisolvens 16S rDNA genes, and of regions 3' and 5' to the genes, was achieved using a linear amplification sequencing reaction with a single oligonucleotide primer, without prior cloning or amplification of the target sequence. The utility of the method was demonstrated by comparison with PCR-amplified template sequencing methods. Sequence differences were revealed by DGS between 16S rRNA genes from two strains of the rumen bacterium Butyrivibrio fibrisolvens isolated in different countries. Using DGS 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.