The isolation and characterization of eight polymorphic microsatellite loci from a Japanese flounder partial genomic library are reported. The eight markers isolated in this study were highly polymorphic and their positions on the linkage genome map of the Japanese flounder were determined. Therefore, they are useful for ecological studies of wild populations. These markers are more effective than other markers with no information of chromosomal locations.
Source and Description. A partial genomic library (300 to 500 bp) was constructed using DNA obtained from Japanese flounder, Paralichthys olivaceus, digested with Sau3AI. The DNA was inserted in pBluescript SK (Toyobo, Japan) digested by BamHI and transformed into competent E. coli cells (strain DH5α). The partial genomic library was screened with a γ 32 P-labeled (CA) 10 probe. About 800 positive clones were obtained, and part of that was sequenced on an ABI Prism 310 Genetic analyzer (Perkin Elmer Japan Applied Biosystems, Tokyo, Japan) using a BigDye Terminator cycle sequencing kit (Perkin Elmer). A total of 150 primer pairs flanking the dinucleotide repeats were designed. A marker, named Poli-RC35-TUF, was found to be highly polymorphic.
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