水牛PNPLA8基因编码区克隆、表达分析及催乳素对该基因的调节作用

doi:10.16431/j.cnki.1671-7236.2020.09.003

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (9): 2715-2723.doi: 10.16431/j.cnki.1671-7236.2020.09.003

水牛PNPLA8基因编码区克隆、表达分析及催乳素对该基因的调节作用

马小娅¹, 庞春英¹, 陆杏蓉¹, 段安琴¹, 梁莎莎¹, 梁燕银², 邓廷贤¹, 梁贤威¹

1. 中国农业科学院广西水牛研究所, 农业农村部(广西)水牛遗传繁殖重点实验室, 南宁 530001;
2. 隆安县动物疫病预防控制中心, 南宁 532700

收稿日期:2020-03-05 出版日期:2020-09-20 发布日期:2020-09-16
通讯作者: 邓廷贤, 梁贤威 E-mail:dtx282000@163.com;liangbri@126.com
作者简介:马小娅(1989-),甘肃白银人,硕士,研究实习员,研究方向:水牛遗传学,E-mail:761195467@qq.com
基金资助:
广西水牛研究所基本科研业务费项目（水牛基180501、水牛基201905007、水牛基201905006、水牛基200513）；广西壮族自治区农业农村厅自筹项目——水牛分子遗传育种创新团队能力建设（Z201948）

Cloning,Expression Analysis and Regulation of PNPLA8 Gene by Prolactin in Buffalo

MA Xiaoya¹, PANG Chunying¹, LU Xingrong¹, DUAN Anqin¹, LIANG Shasha¹, LIANG Yanyin², DENG Tingxian¹, LIANG Xianwei¹

1. Guangxi Key Laboratory of Buffalo Genetics, Breeding and Reproduction, Ministry of Agriculture and Rural Affairs, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning 530001, China;
2. Long'an County Animal Disease Control and Prevention Centre, Nanning 532700, China

Received:2020-03-05 Online:2020-09-20 Published:2020-09-16

摘要/Abstract

摘要： 为了探究磷脂酶patatin样域包含蛋白8（patatin-like phospholipase domain containing 8，PNPLA8）在水牛乳腺脂质代谢中的作用，试验根据GenBank中公布的奶牛PNPLA8基因序列（登录号：XM_005205444.4）设计引物，应用PCR扩增并克隆水牛PNPLA8基因编码区（CDS），应用生物信息学软件分析序列及蛋白质结构；抽提水牛不同组织及不同泌乳期乳腺组织RNA，利用实时荧光定量PCR检测PNPLA8基因在不同组织间和不同泌乳期的表达；利用不同浓度的催乳素处理水牛乳腺上皮细胞，通过定量检测催乳素对PNPLA8基因表达的影响。结果显示，水牛PNPLA8基因CDS长2 355 bp，编码784个氨基酸，与牦牛、山羊等PNPLA8基因具有较高的同源性；PNPLA8基因在所检测的水牛11个组织中有不同水平的表达，在肺脏和乳腺中表达量相对较高，在脂肪和肌肉组织中表达量较低；在整个泌乳期内PNPLA8基因的表达呈现"低-高-低"趋势；催乳素处理水牛乳腺上皮细胞结果显示，随着催乳素浓度升高，PNPLA8基因表达量逐渐下降。本研究成功克隆了水牛PNPLA8基因，并发现PNPLA8基因是参与乳腺泌乳的一个功能基因，为进一步研究PNPLA8基因在水牛乳腺中的功能奠定基础。

关键词: 水牛; PNPLA8基因; 克隆; 泌乳; 乳腺上皮细胞

Abstract: In order to investigate the role of patatin-like phospholipase domain-containing 8(PNPLA8) in lipid metabolism of mammary gland in buffalo,the coding region (CDS) was amplified and cloned by PCR based on the sequence of Bos taurus PNPLA8 gene in GenBank (accession No.:XM_005205444.4) were analyzed using bioinformatics software.Total RNA was extracted from different tissues of buffalo and mammary glands,which were harvested from different lactating buffaloes.The expression of PNPLA8 gene mRNA in different tissues and different lactation period was detected by Real-time quantitative PCR.For buffalo mammary epithelial cell treatment,different concentrations of prolactin were used and the effect of prolactin on the expression of PNPLA8 gene was detected by Real-time quantitative PCR.The results showed that the length of the PNPLA8 gene CDS was 2 355 bp,encoding 784 amino acids.The sequence showed high homology with Bos mutes and Capra hircus.PNPLA8 gene was expressed at different levels in 11 tissues examined,with a relatively high level in the lung and mammary tissues while the low level in the fat and muscle tissues.The expression abundance of the PNPLA8 gene was variable during lactation and showed a trend of "low-high-low".Prolactin treatment showed that the expression of PNPLA8 gene decreased with the increase of prolactin concentration.In this study,PNPLA8 gene of buffalo was successfully cloned,and the expression of PNPLA8 gene in different tissues and the lactation period was analyzed.Herein,the effect of prolactin on the expression of PNPLA8 gene was studied that laid a foundation for further research on PNPLA8 gene of mammary gland in buffalo.

Key words: buffalo; PNPLA8 gene; cloning; lactation; mammary epithelial cell

中图分类号:

S829.9

马小娅, 庞春英, 陆杏蓉, 段安琴, 梁莎莎, 梁燕银, 邓廷贤, 梁贤威. 水牛PNPLA8基因编码区克隆、表达分析及催乳素对该基因的调节作用[J]. 中国畜牧兽医, 2020, 47(9): 2715-2723.

MA Xiaoya, PANG Chunying, LU Xingrong, DUAN Anqin, LIANG Shasha, LIANG Yanyin, DENG Tingxian, LIANG Xianwei. Cloning,Expression Analysis and Regulation of PNPLA8 Gene by Prolactin in Buffalo[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2715-2723.

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水牛PNPLA8基因编码区克隆、表达分析及催乳素对该基因的调节作用

Cloning,Expression Analysis and Regulation of PNPLA8 Gene by Prolactin in Buffalo

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