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http://dx.doi.org/10.5352/JLS.2005.15.6.879

Development of Molecular Detection Marks Using PCR-RFLP Technique for Arkshell (Scapharca broughtonii Schrenck)  

Cho Eun Seob (South Sea Fisheries Research Institute, NFRDI)
lung Choon Coo (South Sea Fisheries Research Institute, NFRDI)
Kim Chul Won (South Sea Fisheries Research Institute, NFRDI)
Sohn Sang Cyu (South Sea Fisheries Research Institute, NFRDI)
Publication Information
Journal of Life Science / v.15, no.6, 2005 , pp. 879-883 More about this Journal
Abstract
This study was differentiated between Korea and China arkshells using PCR-aided RFLP method which could identify the variation for inter-and intra-species of arkshell (Scapharca broughtonii Schrenck) at the level of DNA. The DNA fragment patterns were compared after digesting gene of mitochondrial 16S rDNA with 8 kinds of restriction enzymes. A 720 bp DNA fragment corresponding to 16S rDNA gene was amplified by PCR with primers ArkF-3 and ArkR-3. PCR products were cut by restriction enzymes (Pvull, BamHI, Hinfl, HaeIII, EcoRI, RsaI, Ksp221, and BstX21), and RFLP pattern was studied. A unique 275 bp DNA band was observed in the samples from Dukyang, Gamak, Namhae, Jinhae, and Taean in Korea when treated by Hinfl, but Chinese arkshell did not show. Treatment of HaeIII could discriminate the sample of Namhae and Jinhae from Dukyang/Gamak/Taean, as well as Korean and Chinese arkshell based on a 700 bp. However, PuvII, BamHI, EcoRI, RsaI, Ksp221, and BstX21 showed the same of 700 bp band in Korean and Chinese arkshell. The phylogenetic tree inferred from PCR-RFLP pattern comparsion in Korean arkshell was different that the distance between Dukyang/Gamak/Taean and Namhae/Jinhae was approximately 7. In particular, the distance between Korean and Chinese arkshell was 25. Consequently, HinfI and HaeIII played an important role in a reliable molecular tool for rapid discriminating Korean and Chinese arkshell, as well as a intra-species in Korea.
Keywords
arkshell; molecular tool; PCR-RFLP; restriction enzyme;
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