149

75.牛脊椎畸形複合症(CVM)遺傳缺陷之SSCP 基因型檢測

林德育(1) 曾淑貞(2) 賴永裕(1) 張秀鑾(3) 吳明哲(1)

(1)行政院農業委員會畜產試驗所 (2)中華醫事科技大學 (3)國立屏東科技大學

脊椎畸形複合症（Complex Vertebral Malformation, CVM）是一種乳牛重要的遺傳缺陷，為單一基因突變的顯隱性遺傳。為建立簡易的分子生物學檢測遺傳型的方法，本研究開發CVM 的單股構型多態性DNA片段分析(PCR_SSCP)基因型檢測方法，設計CVM引子組，以血液或牛乳DNA檢體為模板，經PCR 增幅出109 bp的PCR DNA片段，再以SSCP進行電泳染色分析得到不同態樣的基因型。檢測台灣南部一家乳牛場全場458頭乳牛個體血樣DNA，結果有5.5%(25/458)為雜合型(CV, heterozygote)個體，94.5% (433/458)為正常型(TV, test free of CVM)個體，沒有檢出有病型(CVM, affected of CVM)的個體。分析台灣DHI 乳牛群856頭乳牛個別乳DNA樣品，有11.4%(98/856)為雜合型個體，沒有檢出有病型的個體。本試驗開發出第一個可作為監測台灣乳牛群脊椎畸形複合症遺傳缺陷的檢測方法。

關鍵語：乳牛、牛脊椎畸形複合症、遺傳缺陷、單股構型多態性DNA片段分析

SSCP GENOTYPING FOR INHERITED DEFICIENCY OF BOVINE COMPLEX VERTEBRAL MALFORMATION

D. Y. Lin(1), S. J. Tzeng(2), Y. Y. Lai(1), H. L. Chang(3) and M. C. Wu(1)

(1)Livestock Research Institute, Council of Agriculture (2)Chung Hwa University of Medical Technology (3)National Pingtung University of Science and Technology

A method was established to detect complex vertebral malformation (CVM) from blood and milk samples of Holstein dairy cattle in Taiwan using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. CVM gene of dairy cow was amplified 109 bp DNA fragment by polymerase chain reaction (PCR) with CVM primers. Four hundred and fifty eight blood samples of dairy cow from one dairy farm in south Taiwan were genotyped for CVM with the PCR-SSCP method. The frequencies of carrier (CV, heterozygote) and normal (TV, test free of CVM) were 5.5% (25/458) and 94.5% (433/458), respectively. No affect of bovine complex vertebral malformation cattle was detected. There are 11.4% of 856 Holstein dairy cows from Taiwan Dairy Herd Improvement (DHI) were detected as carrier of CVM and no homozygous recessive animal was detected. In the study, we developed a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis for detecting single nucleotide mutations in the candidate genes for CVM, and it was recorded as a first documentation of this genetic deficiency in Taiwan.

Key Words: Dairy cattle, Complex vertebral malformation(CVM), Inherited deficiency, Single-strand conformation polymorphism(SSCP)