Diversity of CRISPR Array and In silico PCR-RFLP Typing Application in Salmonella enterica
DOI:
https://doi.org/10.14456/gag.2018.6Keywords:
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), diversity, Salmonella enterica, In silico PCR-RFLPAbstract
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is the special arrangements of the nucleotide array with alternating between short direct repeat separated by a short spacer sequence. Along with the CRISPR system, they encode an adaptive immune system of bacteria that protects bacterial cell against bacteriophage and plasmid infection. The CRISPR system is found diversely among bacterial species; moreover, these arrays are able to transfer from generation to generation via vertical transfer. However, a survey of the CRISPR arrays diversity in Salmonella enterica isolated from various sources, in order to further contribute to the understanding and application still limit report in Thailand. Here we described two CRISPR-I and CRISPR-II system found in Salmonella enterica, following the analysis of 117 isolates of Salmonella enterica from various sources. The results found that 48 (41%) and 71 (83%) of Salmonella enterica were positive for CRISPR-I and CRISPR-II respectively. The nucleotide analysis shown that the CRISPR-I and CRISPR-II spacer were varying between each isolates, ranged between 2-15 and 1-17 spacers respectively. Moreover, sequencing analysis indicated that the direct repeat region in both CRISPR-I and CRISPR-II has a high diversity. Our study also showed that in silico PCR-RFLP of CRISPR region, and phylogenetic tree construction of the spacers from Salmonella Weltevreden isolated from different sources indicated that it have discrimination ability between intra-serovar. These results imply that the CRISPR array could probably apply as a molecular marker for epidemiological study in Salmonella enterica.References
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