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一种效应即刻早期基因:沼泽水牛和河流水牛中活性调节细胞骨架相关蛋白基因(Arc/Arg3.1)的序列特征、多态性及组织表达谱

Sequence characterization, polymorphism, and tissue expression profile of an effector immediate-early gene: activity-regulated cytoskeletal associated protein gene (Arc/Arg3.1) in swamp and river buffalo.

作者信息

Yuan F, Huo J L, Li D L, Yuan Y Y, Lu W Z, Song S, Li L J, Miao Y W

机构信息

Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan, China.

Domestic Animal Breeding and Crossbreed-Improvement Station of Yunnan Province, Kunming, Yunnan, China.

出版信息

Genet Mol Res. 2014 Mar 31;13(1):2299-309. doi: 10.4238/2014.March.31.10.

DOI:10.4238/2014.March.31.10
PMID:24737478
Abstract

The activity-regulated cytoskeletal associated protein (Arc/Arg3.1) has been implicated in experience-dependent synaptic plasticity and memory formation. However, information regarding its coding gene in buffalo remains scarce. In this study, the full-length of Arc/Arg3.1 was isolated and characterized (accession No. JX491649) and genetic variations of six river buffalo and eight swamp buffalo were investigated. A tissue expression profile was obtained using semi-quantitative reverse transcription-polymerase chain reaction. The coding region sequence of Arc/Arg3.1 contained 1191 nucleotides encoding a putative protein of 396 amino acids with a theoretical isoelectric point (pI) and molecular weight (Mw) of 5.4 and 45.2 kDa, respectively. Four polymorphisms (c.63T>C, c.228T>C, c.558G>A, and c.625G>C) were found in buffalo; however, only substitution c.625G>C was non-synonymous, leading to an amino acid change from Val to Leu at the 209th position of the Arc/Arg3.1 protein sequence. Bioinformatics analysis revealed that this substitution had no significant effect on Arc/Arg3.1 function (subPSEC = -1.4039, Pdeleterious = 0.1685), which indicated that Arc/Arg3.1 was highly conserved and functionally important in buffalo. Phylogenetic analysis revealed that the gene is closely related to that of Bos taurus and Bos grunniens. The gene was moderately expressed in the hypophysis and the placenta; it was weakly expressed in the kidney, milk, mammary gland, cerebrum, lung, heart, rumen, fat, and uterus; and it was almost silent in the muscle, liver, and skin. These findings will provide further insights into the structure and function of the immediate-early gene in buffalo.

摘要

活性调节细胞骨架相关蛋白(Arc/Arg3.1)与经验依赖性突触可塑性和记忆形成有关。然而,关于水牛中其编码基因的信息仍然匮乏。在本研究中,分离并鉴定了Arc/Arg3.1的全长序列(登录号JX491649),并调查了6头河流型水牛和8头沼泽型水牛的基因变异情况。采用半定量逆转录-聚合酶链反应获得了组织表达谱。Arc/Arg3.1的编码区序列包含1191个核苷酸,编码一个推定的由396个氨基酸组成的蛋白质,其理论等电点(pI)和分子量(Mw)分别为5.4和45.2 kDa。在水牛中发现了4个多态性位点(c.63T>C、c.228T>C、c.558G>A和c.625G>C);然而,只有c.625G>C替换是非同义的,导致Arc/Arg3.1蛋白序列第209位的氨基酸从缬氨酸变为亮氨酸。生物信息学分析表明,该替换对Arc/Arg3.1功能无显著影响(subPSEC = -1.4039,Pdeleterious = 0.1685),这表明Arc/Arg3.1在水牛中高度保守且功能重要。系统发育分析表明,该基因与黄牛和牦牛的基因密切相关。该基因在垂体和胎盘中中度表达;在肾脏、乳汁、乳腺、大脑、肺、心脏、瘤胃、脂肪和子宫中弱表达;在肌肉、肝脏和皮肤中几乎不表达。这些发现将为深入了解水牛中即刻早期基因的结构和功能提供进一步的见解。

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