第四十四卷(2015)
乳牛短脊椎綜合症基因檢測法之改進
廖仁寶 陳若菁 吳明哲
行政院農業委員會畜產試驗所
乳牛短脊椎綜合症(BS)為一種隱性遺傳疾病，在 2006 年時首度發現於丹麥。此症狀形成的原 因為在牛第 21 號染色體上的 FANCI 基因有 3.3 kb 片段的缺失。本實驗室初始檢測法條參考與 修改 Fang et al.(2013)所描述者，結果發現該法不但費時且檢測電泳圖像不甚理想。故而進行檢 測方法之改進，於缺失 DNA 片段中設計一引子，採用多重引子 PCR 法，進行乳牛 BS 基因型 檢測，設計 BS 正常基因型者在電泳圖像中 1 kb 處有一條明顯的 DNA 片段，而雜合型者則在 約 0.4 與 1 kb 處各有一條明顯的 DNA 片段。整體而言，新檢測法可比 Fang et al.(2013)使用 法節省 1 小時左右的時間，且將增幅出的 DNA 片段回收定序後，發現該等片段皆為 BS 基因序列。
關鍵語:短脊椎綜合症、基因檢測法、FANCI 基因
IMPROVEMENT OF BRACHISPINA SYNDROME GENOTYPING METHOD FOR DAIRY COWS
R. B. Liaw, J. C. Chen and M. C. Wu
Livestock Research Institute, Council of Agriculture, Executive Yuan
Bovine brachyspina syndrome (BS) is a recessive genetic defect first observed in Denmark in 2006. The syndrome is caused by a 3.3-kb DNA deletion in the bovine Fanconi anemia complementation group I (FANCI) gene on bovine chromosome 21. The preliminary genotyping method in our lab was modified according to the reference published by Fang et al. (2013). However, the result was not good enough to identify BS genotypes clearly. We developed a new method using multiplex PCR. A new primer was designed in the 3.3-kb deleted DNA fragment. By our new genotyping method, the normal genotype will have a clear DNA band at roughly 1 kb. The carriers will have two clear DNA bands at roughly 0.4 and 1 kb. Overall, the new method is 1 hour quicker in genotyping than the one proposed by Fang et al. (2013). Moreover, the method was verified by sequencing the DNA bands recoverd from the target bands on the gel, and it is a better genotyping method than that previous one.
Key Words: Brachyspina syndrome, Genotyping method, FANCI gene
  
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