山羊RPL27A基因克隆及组织表达特性分析

doi:10.16431/j.cnki.1671-7236.2021.11.002

摘要/Abstract

摘要： 为研究山羊核糖体蛋白L27A （ribosome protein L27A，RPL27A）基因结构及其相关的生物学功能，试验采集简州大耳羊的14种组织样品（心脏、肝脏、脾脏、背最长肌、皮下脂肪等），利用RT-PCR扩增并克隆获得山羊RPL27A基因序列，通过在线工具进行生物信息学分析；采用实时荧光定量PCR技术检测RPL27A基因在各个组织及不同分化阶段的皮下前体脂肪细胞中的表达水平。结果显示，山羊RPL27A基因CDS区长447 bp，可编码148个氨基酸。生物信息学分析表明，RPL27A基因在不同的物种间具有较高的保守性，RPL27A蛋白是不稳定的亲水碱性蛋白，其与脂代谢及脂肪沉积相关的RPS18、RPL26、RPL34、RPL32、RPL37A等核糖体蛋白存在相互作用，RPL27A蛋白没有跨膜结构域，且无信号肽，亚细胞定位表明其主要存在于细胞质中。实时荧光定量PCR结果显示，RPL27A基因在山羊心脏、肝脏、脾脏、肾脏、肺脏、背最长肌、皮下脂肪等14种组织中均有广泛表达，且在瘤胃中表达水平最高，在臂三头肌和股二头肌中也存在较高的表达水平，均显著高于其他组织（P<0.05）；RPL27A基因在成脂诱导60 h的山羊皮下脂肪细胞中的表达水平显著高于分化前（P<0.05）。本研究成功克隆了山羊RPL27A基因CDS序列，并揭示了RPL27A基因在山羊14种组织中的表达特性，研究结果可为阐明RPL27A基因对山羊脂肪细胞分化的调控机制提供材料。

关键词: 山羊; RPL27A基因; 克隆; 组织表达

Abstract: In order to study the gene structure and biological function of ribosome protein L27A (RPL27A) in goats, 14 kinds of tissue samples (i. e, heart, liver, spleen, longissimus dorsi muscle, subcutaneous fat, etc.) of Jianzhou Big-eared goats were collected. The sequence of RPL27A gene in goats was obtained by RT-PCR amplification and cloning, and its biological characteristics were analyzed using online tools. Real-time quantitative PCR was used to detect the expression of RPL27A gene in the collected different tissues as well as the subcutaneous preadipocytes at different differentiation stages. The results showed that the CDS region of RPL27A gene in goats was 447 bp, which encoded 148 amino acids. The bioinformatics analysis results showed that RPL27A gene was highly conserved among different species. RPL27A protein was an unstable alkalinity protein, and had obvious interactions with the ribosomal proteins such as RPS18, RPL26, RPL34, RPL32 and RPL37A, which related to lipid metabolism and fat deposition. RPL27A protein had no transmembrane domain and signal peptide, and subcellular localization indicated that it mainly existed in cytoplasm. Real-time quantitative PCR results reveled that RPL27A gene was widely expressed in 14 tissues of goats, such as heart, liver, spleen, kidney, lung, longissimus dorsi muscle and subcutaneous fat, while the expression of RPL27A gene was the highest in rumen, and the expression level in triceps brachii and biceps femoris was significantly higher than that in other tissues (P<0.05). The expression level of RPL27A gene in subcutaneous adipocytes of goats induced for 60 h was significantly higher than that before differentiation (P<0.05). In this study, the CDS sequence of RPL27A gene in goats was successfully cloned, and the expression characteristics of RPL27A gene in 14 tissues of goats were revealed, which could provide materials for elucidating the regulatory effect and mechanism of RPL27A gene on the differentiation of adipocyte in goats.

Key words: goats; RPL27A gene; cloning; tissue expression

中图分类号:

郑建莹, 王金玲, 王永, 朱江江, 张浩. 山羊RPL27A基因克隆及组织表达特性分析[J]. 中国畜牧兽医, 2021, 48(11): 3918-3928.

ZHENG Jianying, WANG Jinling, WANG Yong, ZHU Jiangjiang, ZHANG Hao. Cloning and Tissue Expression Characterization Analysis of RPL27A Gene in Goats[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 3918-3928.

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