Cloning and physical mapping of (GA)n microsatellites in Phalaenopsis stuartiana
Date Issued
2009
Date
2009
Author(s)
Liu, Jung
Abstract
Microsatellites are tandemly repeated tracts of DNA composed of 1–6 base pair long units and ubiquitous in eukaryotic genomes. In this study, fifteen clones riched in (GA)n microsatellites were isolated from Phalaenopsis stuartiana by single-primer PCR using (GA)11 as a primer. All clones were compound except one imperfect repeat sequences. The insert size varied from 28 bp to 726 bp, and the maximum repeat numbers of GA and TC were 28 and 32 in these clones. The sequence identity ranged from 35.98 to 78.57% among them. According to the TIGR database, these sequences showed similarities with repetitive sequences in many plants. Nine clones, eight clones from P. stuartiana and DpGA2 from P. pulcherrima, were selected for physical mapping by fluorescence in situ hybridization (FISH). The signals were clustered at all centromeres of P. stuartiana chromosomes. It was found a quite exceptional distribution of DpGA2 which was clustered at the centromeric regions of only one pair of P. pulcherrima chromosomes. In situ hybridization to P. violacea and P. pulcherrima chromosomes with clone PstGA3 revealed dispersed signals. The result of the former but the latter was consistent with the previous study using synthetic oligonucleotides (GA)11 as a probe. The signals of the clone PstGA16 were scattered along the chromosomes of P. amboienesis, but clustered at the centromeres of P. mannii. It was well correlated with the previous study. Whether (GA)n microsatellite is the functional centromeric element of P. stuartiana remained to resolve by immunostaining or chromatin immunoprecipitation in the future.
Subjects
Phalaenopsis stuartiana
(GA)n microsatellites
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