Identification and Comparison of the Nucleotide Sequence of 16S-23S rRNA Gene Intergenic Small SR(Spacer Region) of Lactobacillus rhamnosus ATCC 53103 with Those of L. casei, L. acidophilus and L. helveticus

  • Byun, J.R. (Department of Animal Science and Technology, Chung-Ang University) ;
  • Yoon, Y.H. (Department of Animal Science and Technology, Chung-Ang University)
  • Received : 2003.03.04
  • Accepted : 2003.07.16
  • Published : 2003.12.01


Reliable PCR based identification of lactobacilli has been described utilizing the sequence of 16S-23S rRNA intergenic spacer region. Those sequence comparisons showed a high degree of difference in homology among the strains of L. rhamnosus, L. casei, L. acidophilus and L. helveticus whose 16S-23S rRNA intergenic small SR's sizes were 222 bp, 222 bp, 206 bp and 216 bp respectively. The sequence of 16S-23S rRNA intergenic spacer region of L. rhamnosus ATCC 53103 revealed the close relatedness to those of L. casei strains by the homology ranges from 95.4% to 97.2%. 16S-23S rRNA intergenic spacer region nucleotide sequence of L. acidophilus showed some distant relatedness with L. rhamnosus ATCC 53103 with the homology ranges from 40.3% to 41.8% and that with L. helveticus was shown to be 30% of homology, which exists at the most distant phylogenetic relatedness. The identification of species and strain of lactobacilli was possible on the basis of these results. The common sequences among the 17 strains were CTAAGGAA located in the initiating position of the DNA and some discrepancies were found between the same strains based on these results.


  1. Loughney, K., E, Lund, and J. E. Dahlberg. 1982. tRNA genes are found between the 16S and 23S genes in Bacillus subtilis. Nucleic Acid Res. 10:1607-1624.
  2. Tannock, G. W., A. Tilsala-Timisjarvi, S. Rodtong, J, NG and K. Munro. 1999. Identification of Lactobacillus isolates from the gastrointestinal Tract, silage, and youghurt by 16S-23S rRNA gene intergenic spacer region sequence comparisons. Appl. Environ. Microbiol. 65(9):4264-4267.
  3. Fox, G.E., J. D Wisottzky and P. Jurtshuk. 1992. How close is close; 16S rRNA sequence identity may not be sufficient to guarantee species identity. Int. J. Syst, Bacteriol. 42, 166-170.
  4. Morieira, D. and R. Amils. 1996. PCR meditated detection of the chemolithotrophic bacterium Thiobacillus cuprinus using 23S rDNA and 16S/23Sintergenic spacer region targeted oligonucleotide primers. FEMS Microbiol. Letter. 142:289-293.
  5. Nakagawa, T., M. Shimada, H. Mukai, K. Asada, I. Kato, K. Fujino and T. Sato. 1994. Detection of alcohol-tolerant hiochi bacteria by PCR. Appl. Environ. Microbiol. 60:637-640.
  6. Yeung, P. S. M., M. E. Sanders, and C. L. Kitts. 2002. Species specific identification of commercial probiotic strains. J. Dairy Sci. 85:1039-1051.
  7. Yoon, Y, H..and B. R. Won. 2002. Antagonism against Helicobacter pylori and proteolysis of Lactobacillus helveticus CU 631 and strain identification. Asian-Aust. J. Anim. Sci. 15(7):1057-1065.
  8. Drake, M. A., C. L. Small, K. D. Spence and B. G. Swanson. 1996. Rapid detection and identification of Lactobacillus spp. in dairy products by using the polymerase chain reaction. J. Food Protec. 59:1031-1036.
  9. Tilsala-Timisjarvi, A. and T. Alatossava. 1997. Development of oligonucleotide primer from the 16S-23S rRNA intergenic sequences for identifying different dairy and probiotic lactic acid bacteria by PCR. Int. J. Food Microbiol. 35:49-56.
  10. Barry, T., G. Colleran, M. Glennon, L. K. Duncann and F. Gannon. 1991. The 16S 23S spacer region as a target for DNA probes to identify eubacteria, PCR methods. Appl.1, 51-56.
  11. Bringel, F., M. C. Curk and J. C. Hubert. 1996. Characterization of lactobacilli by Southern type hybridization with a Lactobacillus plantarum pyr DFE probe Int. J. Syst. Bacteriol. 46:588-594.