Wang Zi-nian, Cai Han-fang, Li Ming-xun, Cao Xiu-kai, Lan Xian-yong, Lei Chu-zhao, Chen Hong
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.
Gene. 2016 Jan 10;575(2 Pt 1):191-8. doi: 10.1016/j.gene.2015.08.047. Epub 2015 Aug 28.
Patatin-like phospholipase domain-containing protein 3 (PNPLA3), a member of the patatin like phospholipase domain-containing (PNPLA) family, plays an important role in energy balance, fat metabolism regulation, glucose metabolism and fatty liver disease. Tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR) is a new method offering fast detection and extreme simplicity at a negligible cost for SNP genotyping. In this paper, we investigated the genetic variations at different ages of 660 Chinese indigenous cattle belonging to three breeds (QC, NY, JX) and applied T-ARMS-PCR and PCR-RFLP methods to genotype four SNPs, SNP1: g.A2980G, SNP2: g.A2996T, SNP3: g.A36718G, SNP4: g.G36850A. The statistical analyses indicated that these 4 SNPs affected growth traits markedly (P<0.05) in QC population, whereas combined haplotypes were not (P>0.05). The qPCR (quantitative PCR) indicated that bovine PNPLA3 gene was exclusively expressed in fat tissues. Besides, the analysis between SNP and mRNA expression revealed that, in SNP1, the expression of AG was much higher than AA and GG (P<0.05), which was in accordance with the results of growth traits association analysis, while the results of SNP4 was not. These results supported high potential that SNPs of bovine PNPLA3 gene might be utilized as genetic markers in marker-assisted selection (MAS) for Chinese cattle breeding programs.
含帕他汀样磷脂酶结构域蛋白3（PNPLA3）是含帕他汀样磷脂酶结构域（PNPLA）家族的成员之一，在能量平衡、脂肪代谢调节、葡萄糖代谢和脂肪肝病中发挥重要作用。四引物扩增阻滞突变系统PCR（T-ARMS-PCR）是一种用于单核苷酸多态性（SNP）基因分型的新方法，具有检测快速、操作极其简单且成本可忽略不计的特点。在本文中，我们调查了属于三个品种（秦川牛、南阳牛、晋南牛）的660头中国本土牛不同年龄阶段的遗传变异情况，并应用T-ARMS-PCR和PCR-RFLP方法对4个SNP进行基因分型，分别为SNP1：g.A2980G、SNP2：g.A2996T、SNP3：g.A36718G、SNP4：g.G36850A。统计分析表明，这4个SNP对秦川牛群体的生长性状有显著影响（P<0.05），而组合单倍型则无显著影响（P>0.05）。定量PCR（qPCR）表明，牛PNPLA3基因仅在脂肪组织中表达。此外，SNP与mRNA表达之间的分析表明，在SNP1中，AG基因型的表达显著高于AA和GG基因型（P<0.05），这与生长性状关联分析结果一致，而SNP4的结果则不然。这些结果支持了牛PNPLA3基因的SNP在我国牛育种计划的标记辅助选择（MAS）中作为遗传标记的巨大潜力。
