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Determination of Enterococcus faecalis groESL Full-Length Sequence and Application for Species Identification
- Source :
- Journal of Clinical Microbiology. 39:3326-3331
- Publication Year :
- 2001
- Publisher :
- American Society for Microbiology, 2001.
-
Abstract
- Amplification of the partial Cpn60 (or GroEL) gene segment has been used for identification of many bacteria, including Enterococcus species. To obtain more sequence data from groESL genes of Enterococcus faecalis , the full-length sequence of the E. faecalis groESL genes containing groES (285 bp), spacer (57 bp), and groEL (1,626 bp) was determined. A database search of GenBank revealed that the deduced E. faecalis GroES and GroEL proteins show significant homology to the GroES and GroEL proteins of other bacteria. The GroEL ( groEL ) of E. faecalis had the highest identity with Streptococcus pneumoniae (81.8% amino acid sequence identity and 73.0% nucleotide sequence identity), followed by Lactococcus zeae , while GroES ( groES ) had 60.2% (64.6%) identity with Lactobacillus zeae and 58.5% (66.2%) identity with Lactococcus lactis , followed by 57.0% (65.5%) identity with Bacillus subtilis. Based on the groES sequence, an E. faecalis -specific PCR assay was developed, and this PCR assay was positive for all the E. faecalis strains tested. Dot blot hybridization using either groES or groEL as the probe distinguished E. faecalis clearly from other species, indicating that both genes can be used as suitable targets for E. faecalis identification. Moreover, broad-range PCR-restriction fragment length polymorphism of groESL was designed to differentiate eight commonly encountered Enterococcus species. The Enterococcus species of reference strains could be easily differentiated on the basis of restriction patterns produced by Hae III and Rsa I. The DNA-based assays developed in this study provide an alternative to currently used methods of identification for clinically important enterococcal species.
- Subjects :
- Microbiology (medical)
Chaperonins
Lactococcus
Molecular Sequence Data
Polymerase Chain Reaction
Enterococcus faecalis
Microbiology
HaeIII
Bacterial Proteins
Species Specificity
medicine
Humans
Cloning, Molecular
Peptide sequence
Gram-Positive Bacterial Infections
biology
Lactococcus lactis
Nucleic acid sequence
Nucleic Acid Hybridization
Bacteriology
Sequence Analysis, DNA
GroES
biology.organism_classification
GroEL
Molecular biology
Bacterial Typing Techniques
Genes, Bacterial
bacteria
Enterococcus
Polymorphism, Restriction Fragment Length
medicine.drug
Subjects
Details
- ISSN :
- 1098660X and 00951137
- Volume :
- 39
- Database :
- OpenAIRE
- Journal :
- Journal of Clinical Microbiology
- Accession number :
- edsair.doi.dedup.....f8e089841578fde8f713ef16dd06b10a