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

Authors:
  • YAYasuo Agawa
    Yasuo Agawa
  • TKTakafumi Komiya
    Takafumi Komiya
  • THTomoki Honryo
    Tomoki Honryo
  • MKMichio Kurata
    Michio Kurata
  • TOTokihiko Okada
    Tokihiko Okada
  • OMOsamu Murata
    Osamu Murata
  • HKHidemi Kumai
    Hidemi Kumai
  • YSYoshifumi Sawada
    Yoshifumi Sawada
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Abstract
PublisherCopyright (c) 2011 日本水産学会, rights: 本文データは学協会の許諾に基づきJ-STAGEから複製したものである, relation: isVersionOf: https://www.jstage.jst.go.jp/article/suisan/77/4/77_4_639/_article/-char/ja/[概要] 太平洋クロマグロ雌雄に特徴的な DNA 断片を検出する目的で AFLP 解析を行った。雌雄 4 個体ずつ計 8 個体よりゲノム DNA を調整し，64 プライマーセットで 2 次増幅した結果 1038 のバンドを検出し，内 183 が個体間で多型を示した。二次増幅を EcoRI+AGG と MseI+CAT プライマーで行った際に雄 32 個体中 29 個体，雌 32 個体中 3 個体において確認される 437 bp の断片を同定した。この断片を定量した結果，雄に於いて検出され易く，また断片の量も検出される雌より有意に（P<0.01）多かった。 [Abstract] 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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