Abstract
Denaturing gradient gel electrophoresis (DGGE) of DNA fragments obtained by PCR amplification of the V2-V3 region of the 16S rRNA gene was used to detect the presence of Lactobacillus species in the stomach contents of mice. Lactobacillus isolates cultured from human and porcine gastrointestinal samples were identified to the species level by using a combination of DGGE and species-specific PCR primers that targeted 16S-23S rRNA intergenic spacer region or 16S rRNA gene sequences. The identifications obtained by this approach were confirmed by sequencing the V2-V3 region of the 16S rRNA gene and by a BLAST search of the GenBank database.
MeSH Terms
Animals
DNA Primers
DNA, Bacterial/analysis
DNA, Ribosomal/analysis
Electrophoresis, Polyacrylamide Gel/methods
Genes, rRNA
Humans
Lactobacillus/classification,genetics,isolation & purification
Mice
Polymerase Chain Reaction/methods
RNA, Ribosomal, 16S/genetics
RNA, Ribosomal, 23S/genetics
Species Specificity
Stomach/microbiology
Chemicals
DNA Primers
DNA, Bacterial
DNA, Ribosomal
RNA, Ribosomal, 16S
RNA, Ribosomal, 23S
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Walter J
Department of Microbiology, University of Otago, Dunedin, New Zealand.
Tannock G W
Tilsala-Timisjarvi A
Rodtong S
Loach D M
Munro K
Alatossava T
References (10)
10 references, click to expand
-
DNA sequencing with chain-terminating inhibitors.
Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7
PMID: 271968
-
Reconstitution of the gastrointestinal microflora of lactobacillus-free mice.
Appl Environ Microbiol. 1988 Dec;54(12):2971-5
PMID: 2975941
-
Basic local alignment search tool.
J Mol Biol. 1990 Oct 5;215(3):403-10
PMID: 2231712
-
Reclassification of Lactobacillus casei subsp. casei ATCC 393 and Lactobacillus rhamnosus ATCC 15820 as Lactobacillus zeae nom. rev., designation of ATCC 334 as the neotype of L. casei subsp. casei, and rejection of the name Lactobacillus paracasei.
Int J Syst Bacteriol. 1996 Jan;46(1):337-40
PMID: 8573516
-
Temperature gradient gel electrophoresis analysis of 16S rRNA from human fecal samples reveals stable and host-specific communities of active bacteria.
Appl Environ Microbiol. 1998 Oct;64(10):3854-9
PMID: 9758810
-
Comparative sequence analyses of the genes coding for 16S rRNA of Lactobacillus casei-related taxa.
Int J Syst Bacteriol. 1997 Jan;47(1):54-7
PMID: 8995801
-
Development of oligonucleotide primers from the 16S-23S rRNA intergenic sequences for identifying different dairy and probiotic lactic acid bacteria by PCR.
Int J Food Microbiol. 1997 Mar 18;35(1):49-56
PMID: 9081225
-
Application of denaturing gradient gel electrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) in microbial ecology.
Antonie Van Leeuwenhoek. 1998 Jan;73(1):127-41
PMID: 9602286
-
Variations of bacterial populations in human feces measured by fluorescent in situ hybridization with group-specific 16S rRNA-targeted oligonucleotide probes.
Appl Environ Microbiol. 1998 Sep;64(9):3336-45
PMID: 9726880
-
Denaturing gradient gel electrophoresis profiles of 16S rRNA-defined populations inhabiting a hot spring microbial mat community.
Appl Environ Microbiol. 1996 Feb;62(2):340-6
PMID: 8593039