Nucleotide sequencing and analysis of 16S rDNA and 16S-23S rDNA internal spacer region (ISR) of Taylorella equigenitalis, as an important pathogen for contagious equine metritis (CEM).
Abstract: The primer set for 16S rDNA amplified an amplicon of about 1500 bp in length for three strains of Taylorella equigenitalis (NCTC11184(T), Kentucky188 and EQ59). Sequence differences of the 16S rDNA among the six sequences, including three reference sequences, occurred at only a few nucleotide positions and thus, an extremely high sequence similarity of the 16S rDNA was first demonstrated among the six sequences. In addition, the primer set for 16S-23S rDNA internal spacer region (ISR) amplified two amplicons about 1300 bp and 1200 bp in length for the three strains. The ISRs were estimated to be about 920 bp in length for large ISR-A and about 830 bp for small ISR-B. Sequence alignment of the ISR-A and ISR-B demonstrated about 10 base differences between NCTC11184(T) and EQ59 and between Kentucky188 and EQ59. However, only minor sequence differences were demonstrated between the ISR-A and ISR-B from NCTC11184(T) and Kentucky188, respectively. A typical order of the intercistronic tRNAs with the 29 nucleotide spacer of 5'-16S rDNA-tRNA(Ile)-tRNA(Ala)-23S rDNA-3' was demonstrated in the all ISRs. The ISRs may be useful for the discrimination amongst isolates of T. equigenitalis if sequencing is employed.
Publication Date: PubMed ID: 16502103
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Summary
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This research investigated the genetic similarities of the Taylorella equigenitalis bacteria, a significant pathogen in contagious equine metritis. The study analyzed the sequence of the 16S rDNA and the internal spacer region (ISR) within the 16S-23S rDNA, highlighting high similarity in the sequences with few differences. The findings also suggest that ISR sequences could be used to effectively distinguish different T. equigenitalis strains.
Introduction and Methodology
- The researchers carried out a detailed examination of a bacteria called Taylorella equigenitalis, which is responsible for causing a contagious equine disease.
- This involved a nucleotide sequencing of two sections of the bacteria’s DNA; firstly the 16S rDNA and secondly the internal spacer region between the 16S rDNA and the 23S rDNA, also known as the ISR.
- They used a primer set for the 16S rDNA which amplified an amplicon approximately 1500 bp (base pairs) long for three different strains of T. equigenitalis. The three strains were NCTC11184(T), Kentucky188, and EQ59.
Detailed Sequencing and Findings
- They found that the 16S rDNA of the six sequences (which included three reference sequences for comparison) were very similar, with only minor differences at a few nucleotide positions.
- The primer set for the ISR also amplified two amplicons around 1300 bp and 1200 bp long for the three examined strains.
- The large ISR-A was approximately 920 bp in length, while the shorter ISR-B was around 830 bp long.
- Sequence alignment of the ISR-A and ISR-B showed that there were around 10 base differences between the NCTC11184(T) and EQ59 strains and between the Kentucky188 and EQ59 strains.
- However, they found only minor differences in the sequences between the ISR-A and ISR-B from NCTC11184(T) and Kentucky188.
Conclusions and Implications
- The area between the 16S rDNA and the 23S rDNA in all the ISR sequences revealed a typical order of the intercistronic tRNAs with a 29 nucleotide spacer; 5′-16S rDNA-tRNA(Ile)-tRNA(Ala)-23S rDNA-3′.
- The findings from this study suggest the ISR region could potentially be used to distinguish different T. equigenitalis strains if sequencing methods are used. This may have important implications for identifying and controlling outbreaks of the equine disease caused by this bacteria.
Cite This Article
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Nucleotide sequencing and analysis of 16S rDNA and 16S-23S rDNA internal spacer region (ISR) of Taylorella equigenitalis, as an important pathogen for contagious equine metritis (CEM).
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Citations
This article has been cited 3 times.- Tazumi A, Sekizuka T, Moore JE, Millar BC, Taneike I, Matsuda M. Molecular characterization of intervening sequences in 23S rRNA genes and 23S rRNA fragmentation in Taylorella equigenitalis. Folia Microbiol (Praha) 2008;53(6):486-92.
- Tazumi A, Ono S, Sekizuka T, Moore JE, Millar BC, Matsuda M. Molecular characterization of the sequences of the 16S-23S rDNA internal spacer region (ISR) from isolates of Taylorella asinigenitalis. BMC Res Notes 2009 Mar 3;2:33.
- Matsuda M, Tazumi A, Kagawa S, Sekizuka T, Murayama O, Moore JE, Millar BC. Homogeneity of the 16S rDNA sequence among geographically disparate isolates of Taylorella equigenitalis. BMC Vet Res 2006 Jan 6;2:1.
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