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57.家禽性染色體上的紡綞蛋白基因分析

楊琇雅 林德育 陳若菁 鍾秀枝 吳明哲

行政院農業委員會畜產試驗所

應用性別鑑定技術來有效繁殖及管理家禽具有極重要的經濟價值，並有助於畜產種原的保存及利用。以土雞、褐色菜鴨、番鴨、白羅曼鵝、青銅火雞、澳洲黑天鵝、鵪鶉(Coturnix coturnix)、環頸雉(Phasianus colchicus)各5公5母個體 DNA作為模版，應用SPIN12及SPIN13引子組進行聚合酶連鎖反應(PCR)後，由電泳分析結果顯示所有受檢物種之雄性與雌性均呈現不同樣態，且與實際性別吻合，證實此引子組可成功鑑別雞、鴨、番鴨、鵝、火雞之性別外，亦可作為黑天鵝、鵪鶉、環頸雉等鳥類的性別遺傳標記，從不同物種所呈現的DNA片段亦有變異存在。本研究以PCR檢測家禽Z與W性染色體中之紡綞體蛋白(Spindlin)基因intron序列，以其長度的差異來鑑定家禽性別，可避免僅檢測W染色體性別鑑定標記的誤判，且不需再搭配內部對照(internal control)，更不需結合限制酶切割法(RFLP)即進行家禽性別鑑定。

關鍵語：家禽、紡綞體蛋白基因、性別鑑定

ANALYSIS OF SPINDLIN GENE IN POULTRY

H. Y. Yang, D. Y. Lin, J. C. Chen, H. C. Chung and M. C. Wu

Livestock Research Institute, Council of Agriculture, Executive Yuan

The importance of sex determination for farm animal production is well known.  To search for DNA-based sex identification candidate marker of poultry, whole blood DNAs from each five males and five females of Native Chicken(Gallus domestics), Brown Tsaiya(Anas platyrhynchos), Muscovy(Cairina moschata), White Roman Goose(Anser cygnoides), Bronze Turkey(Meleagris gallopavo), Australian Black Swan(Cygnus atratus), Quail(Coturnix coturnix) and Ring-necked Pheasant(Phasianus colchicus) were analyzed by spindlin gene.  PCR products were amplified by SPIN12 and SPIN13 primer kits.  Different electrophoresis patterns showed correct gender for each bird species.  The accuracy of sex identification was 100%.  Furthermore, the PCR DNA fragments of the same gender were different among bird species.  Based on different DNA fragments between Z and W chromosome, the developed PCR method has no risk of the mistrial of those sex identification markers depended on W chromosome, and can omit extra internal control or restriction enzyme deigestion.  The DNA-based spindlin gene approach can provide a rapid, exact and low cost method for sex identification in poultry.

Key Words: Poultry, Spindlin gene, Sex identification