技术说明：使用 LightCycler 对绵羊群体中的β-乳球蛋白（CSN2）G 等位基因和κ-乳白蛋白（CSN3）T 等位基因进行常规基因分型的新方法。

Technical note: A novel method for routine genotyping of the G allele of β-casein (CSN2) and T allele of κ-casein (CSN3) in a sheep population using LightCycler.

机构信息

Institute of Animal Science, Department of Molecular Genetics, Prague, Czech Republic.

出版信息

J Anim Sci. 2011 Dec;89(12):3843-5. doi: 10.2527/jas.2011-4054. Epub 2011 Aug 12.

DOI:10.2527/jas.2011-4054

Abstract

The goal of this work was to develop a protocol for rapid genotyping of A and G variants at the CSN2 locus and genotyping of T and C variants at the CSN3 locus in sheep breeds (Sumava and Valachian) by means of PCR and LightCycler analysis. The LightCycler technique combines rapid and efficient in vitro amplification of DNA in glass capillaries with melting curve analysis based on fluorescence resonance energy transfer for the sensitive detection of point mutation. The A variant had a greater frequency (Sumava, 0.778; Valachian, 0.835) than did variant G (Sumava, 0.222; Valachian, 0.165) in both sheep breeds. The CSN3 locus was found to be monomorphic, with no polymorphism identified in either population.

摘要

本研究旨在开发一种通过 PCR 和 LightCycler 分析快速检测绵羊品种（Sumava 和 Valachian）CSN2 基因座 A 和 G 变异以及 CSN3 基因座 T 和 C 变异的方法。LightCycler 技术将玻璃毛细管内的 DNA 快速高效的体外扩增与基于荧光共振能量转移的熔解曲线分析相结合，实现了对点突变的灵敏检测。在这两个绵羊品种中，A 变异的频率均高于 G 变异（Sumava：0.778；Valachian：0.835）。CSN3 基因座在两个群体中均表现为单态性，未发现多态性。