山羊ZFP36L1基因克隆及表达特性分析

Cloning and expression profile of ZFP36L1 gene in goat

摘要:

本研究旨在克隆获取锌指蛋白家族36 (zinc finger protein 36-like 1, ZFP36L1)基因，并明确其表达特性，阐明其在山羊不同组织中的表达模式。采集简州大耳羊的心、肝、脾、肺、肾等15种组织样品，应用逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction, RT-PCR)扩增获得山羊ZFP36L1基因，通过在线工具对基因及蛋白质序列进行生物学分析；再应用实时定量聚合酶链反应(quantitative real-time polymerase chain reaction, qPCR)检测ZFP36L1基因在山羊各个组织及不同分化阶段的肌内前体脂肪细胞中的表达水平，并分析其组织表达谱以及细胞时序表达谱。结果表明，克隆获得山羊ZFR36L1基因序列长度为1 224 bp，编码序列(coding sequence, CDS)区为1 017 bp，编码338个氨基酸，是主要定位于细胞核、细胞质的非分泌不稳定蛋白；组织表达谱显示，ZFP36L1基因在各组织中均有表达，在内脏组织、小肠表达量最高(P<0.01)。在肌肉组织以背最长肌表达量最高(P<0.01)。在脂肪组织中，皮下脂肪组织的表达量显著高于其他组织(P<0.01)。诱导分化结果显示，该基因在肌内前体脂肪细胞成脂分化过程中表达呈上调趋势(P<0.01)。以上研究为阐明山羊ZFP36L1基因的生物学功能提供数据支持。

关键词:山羊;ZFP36L1;基因克隆;肌内前体脂肪细胞;诱导分化

Abstract:

The purpose of this study was to clone and characterize the ZFP36L1 (zinc finger protein 36-like 1) gene, clarify its expression characteristics, and elucidate its expression patterns in different tissues of goats. Samples of 15 tissues from Jianzhou big-eared goats, including heart, liver, spleen, lung and kidney were collected. Goat ZFP36L1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR), then the gene and protein sequence were analyzed by online tools. Quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression level of ZFP36L1 in intramuscular preadipocytes in different tissues and adipocytes of goat at different differentiation stages. The results showed that the length of ZFR36L1 gene was 1 224 bp, and the coding sequence (CDS) region was 1 017 bp, encoding 338 amino acids, which was a non-secretory unstable protein mainly located in nucleus and cytoplasm. Tissue expression profile showed that ZFP36L1 gene was expressed in all selected tissues. In visceral tissues, the small intestine showed the highest expression level (P<0.01). In muscle tissue, the highest expression level was presented in longissimus dorsi muscle (P<0.01), whereas the expression level in subcutaneous adipose tissue was significantly higher than that in other tissues (P<0.01). The results of induced differentiation showed that the expression of this gene was up-regulated during adipogenic differentiation of intramuscular precursor adipocytes (P<0.01). These data may help to clarify the biological function of the ZFP36L1 gene in goat.

Key words:goat;ZFP36L1;gene cloning;intramuscular preadipocyte;induced differentiation