| 牛PLIN1基因CDS区扩增及时序表达 |
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| 引用本文：雷召雄，魏大为，汪书哲，杨梦丽，马 云.牛PLIN1基因CDS区扩增及时序表达[J].西北农业学报,2020,30(10):1472~1478 |
| DOI:10.7606/j.issn.1004-1389.2020.10.003 |
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| 基金项目:国家自然科学基金（31672403）;宁夏回族自治区重点研发引才专项（2019YCZX0068）;宁夏回族自治区科技创新团队计划（03010360052）;现代农业（奶牛）产业技术体系建设项目（CARS-36）。 |
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| 中文摘要:PLIN1（Perilipin1）基因是调控脂类代谢的“分子开关”，其编码的脂滴包被蛋白是脂滴表面含量最丰富的蛋白，在脂肪沉积中起关键作用。采用RT-PCR方法扩增牛PLIN1基因编码区序列，利用组织块法和消化法分离培养牛前体脂肪细胞，进一步利用qPCR技术检测PLIN1在牛皮下前体脂肪细胞诱导分化过程中（0、2、4、6、10 d）mRNA的相对表达水平。结果表明，牛PLIN1基因CDS区长为1 551 bp，共编码517个氨基酸；细胞时序表达结果显示，诱导分化的第0天PLIN1不表达，随着分化时间的推移，其表达量呈上升趋势，与分化的第2天相比，在第6天和第10天表达量上升最快， PPARγ在分化的第0天有少量表达，表达趋势与PLIN1一致，在第6天和第10天显著升高。表明PLIN1在牛脂肪组织生成过程中起重要作用，在前体脂肪细胞分化阶段依赖于 PPARγ的表达而表达。 |
| 中文关键词:牛 PLIN1；基因克隆；时序表达 |
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| Coding Region Sequence and Temporal Expression of Bovine PLIN1 Gene |
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| Abstract:PLIN1 (Perilipin1) is a molecular switch that regulates lipid metabolism, encoded lipid droplet coating protein is the most abundant protein on the surface of lipid droplets and plays a key role in fat deposition. The CDS region of bovine was cloned by RT-PCR，the tissue block method and digestion method were used to isolate and culture bovine preadipocytes ,and the relative mRNA expression level of bovine PLIN1was further detected during subcutaneous preadipocyte differentiation by qPCR (0, 2, 4, 6, 10 d). The results showed that the CDS of bovine PLIN1 gene was 1 551 bp, encoding 517 amino acids. The temporal expression showed that PLIN1 could not expressed on the day 0 during differentiation, but increased with differentiation time. It was significantly increased on the 6 days and 10 days compared with 2 days. PPARγ was also slightly expressed on the day 0 during differentiation, and its expression trend was consistent with PLIN1. In conclusion,PLIN1 plays a key role in lipogenesis of bovine, and its expression depends on PPARγ at the differentiation stage of preadipocyte. |
| keywords:Bovine PLIN1 Gene cloning Temporal expression |
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