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《Chinese Journal of Zoology》 2009-01
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Isolation of Microsatellite Loci from Perca schrenkii and its Universal in Related Species

HAI Sa~ ①② LI Jia-Le~ ① LIU Feng~① FENG Jian-Bin~①(①College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306;② Institute of Fisheries Science, Xinjiang Uygur Autonomous Region, Wulumuqi 830000,China)  
Microsatellites DNA was isolated and enriched from genome of Perca schrenkii by magnetic beads, and were also applied to the other two species in the same genus, i.e. P. fluviatilis and P. flavescens, which aimed to get effective microsatellite primers in the germplasm resources conservation of the species in Perca. Genomic DNA of tail fin from P.schrenkii was extracted and digested with restriction enzyme. Restriction fragments were ligated with linkers and amplified with primers. PCR products were enriched by hybridization with biotin-labeled (CA)_ 15 and biotin-labeled (TG)_ 15 probes. After the selected DNAs amplified, cloned into T/A vectors and transformed into E. coli DH5α, the microsatellite-enriched library of P.schrenkii was successfully constructed. 48 random clones were selected with repeat-sequence primers from the library and were sequenced. As a result, 38 microsatellite sequences were isolated, and the repeat times of motifs in 41 microsatellite loci were much more than 8. 17 pairs of primers were designed from the microsatellite loci all of which were polymorphism for P.schrenkii. These primers were also amplified in P. fluviatilis and P. flavescens, among which 10 pairs could be used, 6 pairs in P. flavescens and 5 in P. flavescens were high polymorphic (PIC0.5).
【Fund】: 上海市重点学科建设项目(Y1101);; 上海高校水产养殖学E-研究院建设项目(03E009)
【CateGory Index】: S917.4
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