Abstract
Paramecium is employed as a valuable model organism in various research fields since a large number of strains with different characteristics of size, morphology, degree of aging, and type of conjugation can be obtained. It is necessary to determine a method for the classification and simple identification of strains to increase their utility as a research tool. This study attempted to establish a polymerase chain reaction (PCR)-based method to differentiate strains of the same species. Genomic DNA was purified from several strains of P. caudatum, P. tetraurelia, and P. bursaria used for comparison by the random amplified polymorphic DNA (RAPD)-PCR method. In P. tetraurelia and P. bursaria, it was sufficiently possible to distinguish specific strains depending on the pattern of random primers and amplification characteristics. For the classification of P. caudatum, based on the sequence data obtained by RAPD-PCR analysis, 5 specific primer sets were designed and a multiplex PCR method was developed. The comparative analysis of 2 standard strains, 12 recommended strains, and 12 other strains of P. caudatum provided by the National BioResource Project was conducted, and specific strains were identified. This multiplex PCR method would be an effective tool for the simple identification of environmental isolates or the management of Paramecium strains.
Highlights
Paramecium is a single-cell free-living eukaryote who lives in freshwaters worldwide, such as ponds and lakes [1]
No 9 (51), one of the standard strains of P. tetraurelia, and No 11 (Yad1g1N), a standard strain of P. bursaria, showed different numbers and patterns of bands compared to other strains (Figs 2 and 3)
The random amplified polymorphic DNA (RAPD) method did not reveal any specific band patterns that could distinguish between syngens and mating types in P. caudatum
Summary
Paramecium is a single-cell free-living eukaryote who lives in freshwaters worldwide, such as ponds and lakes [1]. The artificially cultivation is easy, and it is considered a model organism for many aspects of eukaryotic biology [2,3]. Its high motility is useful in cell biology as a model for studying cell motility [4,5], and its unique and complex reproduction process, including genome duplication and rearrangement, is directly related to its utilization value as a model for sexual reproduction or inheritance analysis [6,7]. Paramecium is a potential host that tolerates the intracellular symbiosis of different organisms [8]. Holospora spp., gram-negative α-proteobacteria, are well known to be maintained within Paramecium nuclei as an obligate symbiont [8,9].
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