牛脊柱畸形综合征检测方法的建立和应用.pdfVIP

HEREDITAS (Beijing) 2008 9 , 30(9): 1223―1227 ISSN 0253-9772 技术与方法 DOI: 10.3724/SP.J.1005.2008.01223 王洪梅, 李建斌, 侯明海, 王长法, 李秋玲, 仲跻峰 , 250100 : 牛脊柱畸形综合征(Complex vertebral malformation, CVM)是近年来新发现的致死性牛常染色体隐性遗传 缺陷病。由于编码UDP-N- 乙酰葡糖胺载体的SLC35A3 基因发生 G →T 的突变而引起本病的发生, 可引起胎牛 死胎、流产、早产。为了解我国正常的荷斯坦牛(黑白花奶牛) 的CVM 携带和发生情况, 建立、应用创造酶切位 点PCR(Created restriction site PCR, CRS-PCR) 、等位基因特异性PCR(Allele-specific polymerase chain reaction, AS-PCR)检测方法检测了表型正常的436 头荷斯坦母牛和93 头荷斯坦公牛, 检测到 3 头CVM 携带者, 其中杂 合母牛 1 头, 杂合公牛2 头, 携带率分别为0.60% 、2.20% 。此方法简便、可靠, 为奶牛CVM 有害基因的分型 和筛选提供了新的方法和思路, 为我国奶牛的分子选育提供了可靠的理论依据。 : 脊柱畸形综合征; 基因突变; AS-PCR; CRS-PCR; 荷斯坦牛 The development and application of method for detecting bovine complex vertebral malformation WANG Hong-Mei, LI Jian-Bin, HOU Ming-Hai, WANG Chang-Fa, LI Qiu-Ling, ZHONG Ji-Feng Dairy Cattle Center of Shandong Academy of Agricultural Sciences, Ji ’nan 250100, China Abstract: Complex vertebral malformation (CVM), a lethal autosomal recessive inherited defect in Holstein calves, was newly reported worldwide. The molecular cause of CVM was a substitution of guanine by thymine (G T) in a solute car- rier family 35 member 3 gene (SLC35A3), encoding UDP-N-acetylglucosamine transporter. It was characterized by stillborn, abortion, and premature birth. The objective of this study was to study the actual carrier frequency of the CVM mutation in a population of Chinese Holstein (=Chinese Black-and-White) normal cattle. The normal 436 Holstein cows and 93 Hol- stein bulls were genotyping by using the Created Restriction Site PCR (CRS-PCR) and Allele-specific PCR (AS-PCR) met