遗传 ›› 2008, Vol. 30 ›› Issue (9): 1223-1227.doi: 10.3724/SP.J.1005.2008.01223

• 技术与方法 • 上一篇    下一篇

牛脊柱畸形综合征检测方法的建立与应用

王洪梅; 李建斌; 侯明海; 王长法; 李秋玲; 仲跻峰

  

  1. 山东省农业科学院奶牛研究中心, 济南 250100

  • 收稿日期:2008-01-04 修回日期:2008-02-26 出版日期:2008-09-10 发布日期:2008-09-10
  • 通讯作者: 王洪梅

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   

  1. Dairy Cattle Center of Shandong Academy of Agricultural Sciences, Ji’nan 250100, China
  • Received:2008-01-04 Revised:2008-02-26 Online:2008-09-10 Published:2008-09-10
  • Contact: WANG Hong-Mei,

摘要:

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。

关键词: 脊柱畸形综合征, 基因突变, CRS-PCR, 荷斯坦牛, AS-PCR

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 carrier 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) methods. There were two bulls and one cow in three observed CVM-carriers. In the Holstein dairy cattle and Holstein bull population, the percentages of CVM carriers were estimated as 0.60% and 2.20% respectively. This study provided a more reliable and useful method for extensive screening of CVM and also offers a theoretical basis for molecular diagnosis in Holstein calves.