Abstract

The NAD⁺-dependent protein deacetylase, Sirtuin3 (SIRT3), plays a role in fertility by preventing the activation of reactive oxygen species (ROS). A novel study was conducted on caprine SIRT3, to study its ovarian expression, explore the sequence variability in exon 7 and analyze its association with prolificacy in two native goat breeds of Kerala, Malabari and Attappady Black. The mRNA isolated from ovaries of six Malabari and Attappady Black goats were subjected to quantitative PCR (qPCR) using GAPDH and β-actin as reference genes. Genomic DNA was isolated from 185 goats (99 Malabari and 86 Attappady Black) and subjected to PCR-SSCP to identify polymorphism in exon 7 of SIRT3 and association with litter size was analyzed. The ovarian expression of caprine SIRT3 was significantly higher (p ≤ 0.01) in Malabari than low prolific Attappady Black. PCR-SSCP analysis revealed, exon 7 of SIRT3 was polymorphic with three genotypes namely, AA, AB and BB with a novel SNP, g.154C > T in the 3′UTR. A significant association (p ≤ 0.05) was noticed between the genotypes of SIRT3 and litter size. The results obtained from this study highlight the role of SIRT3 in reproduction and hence SIRT3 may be considered as a potential candidate gene for genetic improvement in goats.

摘要

NAD ⁺依赖性蛋白脱乙酰酶 Sirtuin3 (SIRT3) 通过阻止活性氧 (ROS) 的激活在生育中发挥作用。对山羊SIRT3进行了一项新研究，以研究其卵巢表达，探索外显子 7 的序列变异性，并分析其与喀拉拉邦、马拉巴里和阿塔帕迪黑两种本地山羊品种的多产的关联。使用GAPDH和β-肌动蛋白作为参考基因，对从六只马拉巴里山羊和阿塔帕迪黑山羊的卵巢中分离的 mRNA 进行定量 PCR (qPCR) 。从 185 只山羊（99 只马拉巴里山羊和 86 只 Attappady Black）中分离基因组 DNA，并进行 PCR-SSCP 以鉴定SIRT3外显子 7 的多态性并分析了与窝产仔数的关联。 在马拉巴里，山羊SIRT3的卵巢表达显着高于低产的 Attappady Black （p ≤ 0.01）。PCR-SSCP 分析显示，SIRT3 的外显子 7是多态性的，具有 AA、AB 和 BB 三种基因型，具有新的 SNP，3'UTR 中的 g.154C > T。 注意到SIRT3基因型和窝产仔数之间存在显着关联 ( p ≤ 0.05) 。从这项研究中获得的结果突出了SIRT3在繁殖中的作用，因此SIRT3可能被认为是山羊遗传改良的潜在候选基因。