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山羊黏多醣症遺傳缺陷之SSCP 基因型檢測

林德育(1) 黃鈺嘉(1) 陳若菁(1) 張秀鑾(2) 吳明哲(1)

(1)行政院農業委員會畜產試驗所 (2) 國立屏東科技大學畜產系

本研究建立山羊黏多醣症遺傳缺陷之單股構型多態性 (SSCP) 基因型檢測方法，應用此方法來檢測乙醯醣胺氨基硫酸酶 (N-acetylglucosamine 6-sulfatase, G6S)基因型不再需要使用限制酶，可節省檢測時間、成本與人力。G6S基因的遺傳變異在山羊黏多醣症第三型 (Mucopolysaccharidosis IIID)扮演重要的角色。以限制酶分析該基因所增幅出的聚合酶連鎖反應的產物已被應用於該遺傳疾病的檢測，其突變點的位置在G6S cDNA 第322個核苷酸上 (C→T)。聚合酶連鎖反應結合單股構型多態性分析 (PCR-SSCP) 是一種應用於鑑別基因 DNA 序列上的突變或多態性的技術，在針對 DNA 序列的點突變與多態性的分析上較聚合酶連鎖反應結合限制酶切割片段長度多型性分析(PCR-RFLP) 更有利。發展出來的山羊黏多醣症遺傳缺陷之 SSCP 基因型檢測方法，可以快速有效率的檢測 G6S 基因型，以此法檢測 49 頭努比亞山羊，結果與 PCR-RFLP 的檢測結果一致 (四頭雜合型與45頭正常型)，可以準確地檢測出山羊黏多醣症。

關鍵語：山羊、單股構型多態性、黏多醣症。

SSCP GENOTYPING FOR INHERITED DEFICIENCY OF
CAPRINE MUCOPOLYSACCHARIDOSIS IIID

D. Y. Lin(1), Y. C. Huang(1), J. C. Chen(1), H. L. Chang(2) and M. C. Wu(1)

(1)Livestock Research Institute, Council of Agriculture
(2)Department of Animal Science, National Pingtung University of Science and Technology

Genetic variation in the N-acetylglucosamine 6-sulfatase (G6S) gene is the key role in caprine Mucopolysaccharidosis IIID. A more efficient SSCP (single-strand conformation polymorphism) method is experimented to identify goat G6S genotypes, allowing the detection of the sequence variation without using endonuclease. Comparing the reported G6S PCR-RFLP method, which applied the restriction site around the 322 mutation position (C→T) of N-acetylglucosamine 6-sulfatase cDNA. Instead, with new designed primers, the updating PCR-SSCP was one of the efficient alternatives for identifying G6S mutant genotypes. From the PCR-SSCP tested results of 49 blood samples of Nubian goats, there were four carrier and 45 normal, which was consisted with PCR-RFLP’s. The advantage of the improved goat G6S SSCP over RFLP analysis was having minimal number of steps, which can save time and total cost with acceptable accuracy.　

Key Words: Goat, Single-strand conformation polymorphism(SSCP), Mucopolysaccharidosis.