敲降VPS28基因对中国荷斯坦奶牛乳脂合成的调控

doi:10.16288/j.yczz.18-134

遗传 ›› 2018, Vol. 40 ›› Issue (12): 1092-1100.doi: 10.16288/j.yczz.18-134

敲降VPS28基因对中国荷斯坦奶牛乳脂合成的调控

刘莉莉¹,郭爱伟¹,吴培福¹,陈粉粉¹,杨亚晋¹,张勤²()

^1. 西南林业大学生命科学学院,昆明 650224
^2. 中国农业大学动物科技学院,畜禽育种国家工程实验室,北京 1000193

收稿日期:2018-05-15 修回日期:2018-07-31 出版日期:2018-12-20 发布日期:2018-10-30
通讯作者: 张勤 E-mail:qzhang@cau.edu.cn
作者简介:刘莉莉,博士,讲师,研究方向：动物遗传育种。E-mail: liulily0518@163.com
基金资助:
国家自然科学基金项目(3120772);国家转基因育种发展专项(2014ZX0800953B);教育部重点学科生物学一级学科建设项目(51400666);云南省教育厅科学研究基金教师类项目(2018JS333)

Regulation of VPS28 gene knockdown on the milk fat synthesis in Chinese Holstein dairy

Liu Lili¹,Guo Aiwei¹,Wu Peifu¹,Chen Fenfen¹,Yang Yajin¹,Zhang Qin²()

^1. College of Life Science, Southwest Forestry University, Kunming 650224, China
^2. Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture,College of Animal Science and Technology, China Agricultural University, Beijing 100193, China

Received:2018-05-15 Revised:2018-07-31 Online:2018-12-20 Published:2018-10-30
Contact: Zhang Qin E-mail:qzhang@cau.edu.cn
Supported by:
Supported by the National Natural Science Foundation of China(3120772);the National Major Development Program of Transgenic Breeding(2014ZX0800953B);Ministry of education key subject biology level discipline construction project(51400666);Yunnan Provincial Department of Education Fund for Scientific Research(2018JS333)

摘要/Abstract

摘要：

本课题组前期通过GWAS研究,发现VPS28基因在荷斯坦奶牛乳腺组织中特异性高表达,且其5′-UTR的突变位点-58C>T与乳脂性状关联,但其对乳脂性状的调控机理尚未明确。本研究为了明确VPS28基因及其突变位点-58C>T对乳脂的调控机理,首先利用启动子活性分析检测突变位点-58C>T对VPS28基因的影响,发现该突变位点显著降低VPS28基因启动子活性;然后利用RNA干扰技术敲降奶牛原代乳腺上皮细胞中VPS28基因表达量,检测VPS28通路和乳脂合成相关基因mRNA表达量以及细胞中脂肪滴形态,分析结果发现敲降VPS28基因可降低泛素化-溶酶体和泛素化-蛋白酶体通路基因和乳脂合成相关基因的表达量,并提高细胞中甘油三酯的合成,预示VPS28基因可能通过泛素化-溶酶体和泛素化-蛋白酶体途径调控乳脂生成。本研究结果在转录组水平揭示VPS28基因对乳脂合成的调控机制,为奶牛乳脂性状的分子育种研究提供参考依据。

关键词: 中国荷斯坦奶牛, VPS28基因, 单核苷酸多态性, 乳脂合成, 调控

Abstract:

In a previous genome-wide association study on milk production traits in Chinese Holstein population, we discovered VPS28 gene was highly expressed in mammary gland tissue. Further, a -58C>T mutantion at the 5°-UTR of the gene was significantly associated with milk fat content traits. However, its regulation on milk fat content is still uncertain. In this study, we explored the effect of this -58C>T mutation on VPS28, and found that it could significantly reduce promoter activity of VPS28 gene. To identify the potential function of this SNP, we performed RNAi experiments to knockdown VPS28 gene in BMECs and examined the general effects of VPS28 knockdown on BMECs. The results showed that VPS28 knockdown could decrease the mRNA expression of genes in ubiquitination-lysosome and ubiquitination- proteasome pathways, increase the mRNA expression of genes in milk fat synthesis pathway and promote triglyceride synthesis in BMECs. Our data indicate that VPS28 gene could regulate milk fat synthesis pathway; and promote triglyceride synthesis in BMECs. Our data indicate that VPS28 gene could regulate milk fat synthesis through modulating the ubiquitination-lysosome and ubiquitination-proteasome systems. Our results demonstrate the molecular mechanism of VPS28 on regulation of milk synthesis in dairy cattle through the ubiquitination signaling pathway, thereby supporting a relationship between milk fat synthesis and ubiquitination and laying the molecular foundation in breeding of dairy cattle in the future.

Key words: Chinese Holstein dairy, VPS28 gene, single nucleotide polymorphism, milk fat related genes, transcriptional regulation

刘莉莉, 郭爱伟, 吴培福, 陈粉粉, 杨亚晋, 张勤. 敲降VPS28基因对中国荷斯坦奶牛乳脂合成的调控[J]. 遗传, 2018, 40(12): 1092-1100.

Liu Lili, Guo Aiwei, Wu Peifu, Chen Fenfen, Yang Yajin, Zhang Qin. Regulation of VPS28 gene knockdown on the milk fat synthesis in Chinese Holstein dairy[J]. Hereditas(Beijing), 2018, 40(12): 1092-1100.

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敲降VPS28基因对中国荷斯坦奶牛乳脂合成的调控

Regulation of VPS28 gene knockdown on the milk fat synthesis in Chinese Holstein dairy

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