The feasibility of intragenerically characterizing bifidobacteria by a comparison of a short region within the recA gene was tested. An ~ 300 bp fragment of the recA gene was PCR-amplified from six species from the genus Bifidobacterium using primers directed to two universally conserved regions of the recA gene. A phylogenetic analysis of the sequenced I recA products compared favorably to classification based on the 16S rRNA sequences of the species tested. To apply this rapid methodology to unknown human intestinal bifidobacteria, 46 isolates were randomly chosen from the feces of four subjects and initially characterized by RFLP analysis of a PCR-amplified region of their 16S RNA genes. From a representative of the dominant RFLP family in each of the subjects, the recA segment was PCR-amplified, sequenced and phylogenetically analyzed. All four isolates were found to be related to one another and to B. longum and B. infantis. These results illustrate that the r recA gene may be useful for intrageneric phylogenetic analysis as well as for the identification of unknown fecal bifidobacteria.
Bibliographical noteFunding Information:
This study was supported in part by Minnesota Agricultural Experiment Station (Projects MN-18-055 and MN-18-069), Minnesota South Dakota Dairy Foods Research Center and Systems Bio-Industries Inc.
- 16S ribosomal RNA
- Gastrointestinal tract