Limits...
Estimation of 16S rRNA gene copy number in several probiotic Lactobacillus strains isolated from the gastrointestinal tract of chicken.

Lee CM, Sieo CC, Abdullah N, Ho YW - FEMS Microbiol. Lett. (2008)

Bottom Line: Genomic DNA of the strains was digested with restriction endonucleases that do not cut within the 16S rRNA gene of the strains.The copy number of the 16S rRNA gene within a Lactobacillus species was found to be conserved.From the hybridization results, Lactobacillus salivarius I 24 was estimated to have seven copies of the 16S rRNA gene, Lactobacillus panis C 17 to have five copies and Lactobacillus gallinarum strains I 16 and I 26 four copies.

View Article: PubMed Central - PubMed

Affiliation: Microbial Biotechnology Unit, Laboratory of Industrial Biotechnology, Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.

ABSTRACT
The copy numbers of 16S rRNA genes in 12 probiotic Lactobacillus strains of poultry origin were analyzed. Genomic DNA of the strains was digested with restriction endonucleases that do not cut within the 16S rRNA gene of the strains. This was followed by Southern hybridization with a biotinylated probe complementary to the 16S rRNA gene. The copy number of the 16S rRNA gene within a Lactobacillus species was found to be conserved. From the hybridization results, Lactobacillus salivarius I 24 was estimated to have seven copies of the 16S rRNA gene, Lactobacillus panis C 17 to have five copies and Lactobacillus gallinarum strains I 16 and I 26 four copies. The 16S rRNA gene copy numbers of L. gallinarum and L. panis reported in the present study are the first record. Lactobacillus brevis strains I 12, I 23, I 25, I 211, I 218 and Lactobacillus reuteri strains C 1, C 10, C 16 were estimated to have at least four copies of the 16S rRNA gene. In addition, distinct rRNA restriction patterns which could discriminate the strains of L. reuteri and L. gallinarum were also detected. Information on 16S rRNA gene copy number is important for physiological, evolutionary and population studies of the bacteria.

Show MeSH
Southern blot analysis of the 16S rRNA gene of Lactobacillus reuteri C 1 (a), C 16 (b) and C 10 (c). Several enzymes were used to digest genomic DNA of the Lactobacillus strains. Lane 1, BamHI; lane 2, BclI; lane 3, EcoRI; lane 4, EcoRV; lane 5, HindIII; lane 6, NheI; lane 7, SalI.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2613232&req=5

fig05: Southern blot analysis of the 16S rRNA gene of Lactobacillus reuteri C 1 (a), C 16 (b) and C 10 (c). Several enzymes were used to digest genomic DNA of the Lactobacillus strains. Lane 1, BamHI; lane 2, BclI; lane 3, EcoRI; lane 4, EcoRV; lane 5, HindIII; lane 6, NheI; lane 7, SalI.

Mentions: Digestion of chromosomal DNA of L. reuteri C 1, C 10 and C 16 with BamHI, BclI, EcoRI and EcoRV yielded four hybridizing bands of varying intensities, indicating the existence of at least four copies of the 16S rRNA gene (Fig. 5). It was also observed that the hybridizing patterns of these digests and those of HindIII, NheI and SalI for L. reuteri C 1 and C 16 were identical (Fig. 5a and b). The full genome sequencing of L. reuteri DSM 20016T (GenBank accession number CP000705) was completed recently and it was found that the 16S rRNA gene was present in six copies. The discrepancy between this result and that of the present study suggests that the hybridizing bands with strong signals in the present study could contain more than one 16S rRNA gene.


Estimation of 16S rRNA gene copy number in several probiotic Lactobacillus strains isolated from the gastrointestinal tract of chicken.

Lee CM, Sieo CC, Abdullah N, Ho YW - FEMS Microbiol. Lett. (2008)

Southern blot analysis of the 16S rRNA gene of Lactobacillus reuteri C 1 (a), C 16 (b) and C 10 (c). Several enzymes were used to digest genomic DNA of the Lactobacillus strains. Lane 1, BamHI; lane 2, BclI; lane 3, EcoRI; lane 4, EcoRV; lane 5, HindIII; lane 6, NheI; lane 7, SalI.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2613232&req=5

fig05: Southern blot analysis of the 16S rRNA gene of Lactobacillus reuteri C 1 (a), C 16 (b) and C 10 (c). Several enzymes were used to digest genomic DNA of the Lactobacillus strains. Lane 1, BamHI; lane 2, BclI; lane 3, EcoRI; lane 4, EcoRV; lane 5, HindIII; lane 6, NheI; lane 7, SalI.
Mentions: Digestion of chromosomal DNA of L. reuteri C 1, C 10 and C 16 with BamHI, BclI, EcoRI and EcoRV yielded four hybridizing bands of varying intensities, indicating the existence of at least four copies of the 16S rRNA gene (Fig. 5). It was also observed that the hybridizing patterns of these digests and those of HindIII, NheI and SalI for L. reuteri C 1 and C 16 were identical (Fig. 5a and b). The full genome sequencing of L. reuteri DSM 20016T (GenBank accession number CP000705) was completed recently and it was found that the 16S rRNA gene was present in six copies. The discrepancy between this result and that of the present study suggests that the hybridizing bands with strong signals in the present study could contain more than one 16S rRNA gene.

Bottom Line: Genomic DNA of the strains was digested with restriction endonucleases that do not cut within the 16S rRNA gene of the strains.The copy number of the 16S rRNA gene within a Lactobacillus species was found to be conserved.From the hybridization results, Lactobacillus salivarius I 24 was estimated to have seven copies of the 16S rRNA gene, Lactobacillus panis C 17 to have five copies and Lactobacillus gallinarum strains I 16 and I 26 four copies.

View Article: PubMed Central - PubMed

Affiliation: Microbial Biotechnology Unit, Laboratory of Industrial Biotechnology, Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.

ABSTRACT
The copy numbers of 16S rRNA genes in 12 probiotic Lactobacillus strains of poultry origin were analyzed. Genomic DNA of the strains was digested with restriction endonucleases that do not cut within the 16S rRNA gene of the strains. This was followed by Southern hybridization with a biotinylated probe complementary to the 16S rRNA gene. The copy number of the 16S rRNA gene within a Lactobacillus species was found to be conserved. From the hybridization results, Lactobacillus salivarius I 24 was estimated to have seven copies of the 16S rRNA gene, Lactobacillus panis C 17 to have five copies and Lactobacillus gallinarum strains I 16 and I 26 four copies. The 16S rRNA gene copy numbers of L. gallinarum and L. panis reported in the present study are the first record. Lactobacillus brevis strains I 12, I 23, I 25, I 211, I 218 and Lactobacillus reuteri strains C 1, C 10, C 16 were estimated to have at least four copies of the 16S rRNA gene. In addition, distinct rRNA restriction patterns which could discriminate the strains of L. reuteri and L. gallinarum were also detected. Information on 16S rRNA gene copy number is important for physiological, evolutionary and population studies of the bacteria.

Show MeSH