应用MGB荧光探针技术确定猪KIT基因拷贝重复数

畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (3): 437-441.doi:

应用MGB荧光探针技术确定猪KIT基因拷贝重复数

王韬，唐辉*，曾勇庆，李晓宁，闫超，董林松，于希江*

山东农业大学动物科技学院，泰安 271018

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-03-25 发布日期:2011-03-25
通讯作者: 唐辉;于希江

Detecting Copy Number Variations in Porcine KIT by TaqMan-MGB Probe

WANG Tao, TANG Hui* , ZENG Yong-qing, LI Xiao-ning, YAN Chao,
DONG Lin-song, YU Xi-jiang*

College of Animal Science & Technology, Shandong Agricultural University,Tai′an 271018, China

Received:1900-01-01 Revised:1900-01-01 Online:2011-03-25 Published:2011-03-25

摘要/Abstract

摘要： 旨在建立一种简便、快速检测猪KIT基因拷贝重复数（CNVs）的方法并应用于未知样品的检测。在KIT外显子2中设计TaqMan-MGB探针和引物，建立荧光实时定量PCR检测KIT基因CNVs的方法。结果表明，TaqMan-MGB探针扩增KIT基因目的片段，不同DNA梯度浓度Ct值差异极显著（P<0.001），变异系数低（0.12%～0.26%）；KIT与内标基因ESR的扩增效率相差只有2.4%；采用聚类分析推测了待测的50枚样品KIT的CNVs分别归属于2、3、4、5、6个拷贝。本研究建立的TaqManMGB探针实时荧光定量PCR检测KIT基因CNVs的方法，具有简便、快速、特异性和稳定性好的特点，适合于普通实验室应用。

Abstract: The main objective of the present study was to establish a simple, rapid method for detecting copy number variations (CNVs) in porcine KIT gene TaqMan-MGB probes and primers were designed according to sequence of exon 2 of KIT gene, and a real-time fluorescence quantitative PCR procedure was established for the CNVs detection. The results showed that amplification curves of KIT obtained by TaqMan-MGB probe were a series of parallel curves corresponding to 2-fold serial dilutions of DNA samples, Ct values between groups were significantly different (P<0.001), and the coefficient-of-variations were low (from 0.12% to 0.26%). The amplification efficiencies of the KIT and ESR were approximately equal. The CNVs in KIT of 50 pigs were estimated by cluster analysis, assigned to 2, 3, 4, 5 or 6 copies, respectively. The real-time quantitative PCR using TaqMan-MGB probe is a simple, rapid method with high resolution and stability to measure CNVs in KIT, and it could be carried out in common laboratories.

王韬;唐辉;曾勇庆;李晓宁;闫超;董林松;于希江. 应用MGB荧光探针技术确定猪KIT基因拷贝重复数[J]. 畜牧兽医学报, 2011, 42(3): 437-441.

WANG Tao;TANG Hui;ZENG Yong-qing;LI Xiao-ning;YAN Chao;DONG Lin-song;YU Xi-jiang . Detecting Copy Number Variations in Porcine KIT by TaqMan-MGB Probe[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2011, 42(3): 437-441.