黑天鵝性別鑑定的RAPD標記

林德育 劉瑞珍 陳若菁 吳國欽 葉力子 張秀鑾

行政院農業委員會畜產試驗所

為尋求黑天鵝性別遺傳標記，本研究應用逢機複製多態性 DNA (RAPD)分析方法，以行政院農業委員會畜產試驗所彰化種畜繁殖場所選育的黑天鵝個體全血 DNA，進行性別遺傳標記探討。第一階段以6 公6 母黑天鵝個體 DNA作為模版，共使用220組寡核苷酸 RAPD 引子，進行RAPD分析，尋找區分性別的後選遺傳標記。分析結果顯示 220 組寡核苷酸引子中，引子 AE03(5’- CATAgAgCgg -3’) 所產出的 PCR 產物之 690 bp DNA 片段為雌黑天鵝特有的片段，可作為區分性別的標記。第二階段，再以引子 AE03 分析該場2001年全場58隻黑天鵝DNA樣本，檢測結果顯示DNA判讀之性別均與與實際性別吻合，準確率為 100% (58/58)；意即所有檢測雌黑天鵝PCR 產物 DNA 片段中均具有690 bp DNA 片段，而在所有受檢雄黑天鵝則無，證實 RAPD 引子 AE03 可作為區分性別之遺傳標記。

關鍵語：黑天鵝、性別鑑定、逢機複製多態性 DNA。

RAPD MARKER FOR SEX IDENTIFICATION OF BLACK SWAN

D. Y. Lin, J. J. L. Tai, J. C. Chen, G.. C. Wu, L. T. Yeh and H. L. Chang

Livestock Research Institute, Council of Agriculture, Executive Yuan

To search for Black Swan sex identification candidate markers, whole blood DNA of individual Black Swan from Changhua Animal Propagation Station of Livestock Research Institute, Council of Agriculture (LRI, COA) were analyzed by.random amplified polymorphic DNA (RAPD). In phase I, 220 different 10-mer primers of arbitrary sequence were used to screen twelve Black Swan (6 males and 6 females) in 1996. A RAPD marker (AE03, 5’- CATAgAgCgg -3’) demonstrated high sensitivity and accuracy in gender diagnosis. The PCR polymorphism results indicated a specific DNA fragment (690 bp) amplified with primer AE03 could distinguish females from males. The proof stage, phase II, AE03 was used to test other 58 birds(28 males and 30 females) from the Station in Sep. 2001. Results showed 100% (58/58) accurate. The specific DNA fragment (690 bp) amplified with primer AE03 could be a sex identification marker of Black Swan.

Key words: Black Swan, Sexing, Random amplified polymorphic DNA (RAPD).