Nippon Suisan Gakkaishi 77 (4), 639-646 (2011)

Screening of the male characteristic DNA fragment using AFLP analysis in Pacific bluefin tuna, Thunnus orientalis

YASUO AGAWA,1* TAKAFUMI KOMIYA,1 TOMOKI HONRYO,1 MICHIO KURATA,2
TOKIHIKO OKADA,2 OSAMU MURATA,3 HIDEMI KUMAI4 AND YOSHIFUMI SAWADA1

Fisheries Laboratories, Kinki University, 1Oshima Experiment Station, 2Oshima Fish Nursery Center, Kushimoto, Wakayama 649-3633, 3Shirahama Experiment Station, Shirahama, Wakayama 649-2211, 4Uragami Experiment Station, Nachikatsuura, Wakayama 649-5145, Japan

AFLP, amplified fragment length polymorphism screening, was conducted to identify sex characteristic DNA markers of cultured Pacific bluefin tuna (PBT), Thunnus orientalis. The AFLP screening of 4 females and 4 males with 64 primer combinations produced a total of 1038 scorable bands, of which 183 were polymorphic between individuals. Male characteristic fragments were confirmed with EcoRI+AGG and MseI+CAT primers by gel image analysis, which corresponded to 437 bp in 29 of 32 males and 3 of 32 females. Fragment detail was analyzed by electropherogram analysis and DNA quantification. The 437 bp fragment was detected in 30 individuals of average DNA concentration 0.231 ng/μL; while in females, 13 individuals were detected with average 0.080 ng/μL (P<0.01).


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