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四引物扩增阻滞突变系统PCR(T-ARMS-PCR)快速鉴定出影响生长性状的牛ACADVL基因关键错义突变(P236T)。

Tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR) rapidly identified a critical missense mutation (P236T) of bovine ACADVL gene affecting growth traits.

作者信息

Zhang Sihuan, Dang Yonglong, Zhang Qingfeng, Qin Qiaomei, Lei Chuzhao, Chen Hong, Lan Xianyong

机构信息

College of Animal Science and Technology, Northwest A&F University, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China.

College of Animal Science and Technology, Northwest A&F University, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China.

出版信息

Gene. 2015 Apr 1;559(2):184-8. doi: 10.1016/j.gene.2015.01.043. Epub 2015 Jan 22.

Abstract

Acyl-CoA dehydrogenase, very long chain (ACADVL), encoding ACADVL protein, targets the inner mitochondrial membrane where it catalyzes the first step of the mitochondrial fatty acid beta-oxidation pathway and plays an important role in body metabolism and oxidation of long chain fatty acid releasing energy. Tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR) is an easy-to-operate, rapid, inexpensive, and exact method for SNP genotyping. Herein, T-ARMS-PCR was carried out to detect a critical missense mutation (AC_000176:g.2885C>A; Pro236Thr) within the ACADVL gene in 644 individuals from two cattle breeds. In order to evaluate the accuracy of the T-ARMS-PCR at this locus, the genotype of the sampled individuals was also identified by PCR-RFLP. The concordance between these two methods was 98.76%. Statistical analysis showed that the bovine ACADVL gene had a significant effect on chest width (P<0.05), chest depth (P<0.05), and hip width (P<0.05) in the Qinchuan breed. The cattle with AA genotype had superior growth traits compared to cattle with AC and/or CC genotypes. The "A" allele had positive effects on growth traits. Therefore, T-ARMS-PCR can replace PCR-RFLP for rapid genotyping of this mutation, which could be used as a DNA marker for selecting individuals with superior growth traits in the Qinchuan breed. These findings contribute to breeding and genetics in beef cattle industry.

摘要

酰基辅酶A脱氢酶超长链(ACADVL)编码ACADVL蛋白,定位于线粒体内膜,在那里催化线粒体脂肪酸β-氧化途径的第一步,在机体代谢以及长链脂肪酸氧化释放能量过程中发挥重要作用。四引物扩增阻滞突变系统PCR(T-ARMS-PCR)是一种操作简便、快速、廉价且准确的单核苷酸多态性(SNP)基因分型方法。在此,运用T-ARMS-PCR对两个牛品种的644个个体的ACADVL基因中的一个关键错义突变(AC_000176:g.2885C>A;Pro236Thr)进行检测。为评估T-ARMS-PCR在该位点的准确性,还通过PCR-RFLP对采样个体的基因型进行鉴定。这两种方法的一致性为98.76%。统计分析表明,秦川牛品种中,牛ACADVL基因对胸围(P<0.05)、胸深(P<0.05)和臀围(P<0.05)有显著影响。与AC和/或CC基因型的牛相比,AA基因型的牛具有更优的生长性状。“A”等位基因对生长性状有积极影响。因此,T-ARMS-PCR可替代PCR-RFLP对该突变进行快速基因分型,其可作为一个DNA标记用于选择秦川牛品种中具有优良生长性状的个体。这些研究结果对肉牛产业的育种和遗传学研究有重要意义。

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