荷斯坦奶牛DPP6和PRKN基因的生物信息学及表达规律分析  

Bioinformatics and tissue expression analysis of DPP6 and PRKN in Holstein

作　　者：李瑞瑞 虎红红 冯雪 杨文飞 马云 LI Rui-rui;HU Hong-hong;FENG Xue;YANG Wen-fei;MA Yun(College of Animal Science and Technology,Ningxia University/Ningxia Hui Autonomous Region Key Laboratory of Ruminant Molecular Cell Breeding,Yinchuan 750021,China;Ningxia Xinao Agriculture and Animal Husbandry Co.,Ltd.,Lingwu,Ningxia 750406,China)

机构地区：[1]宁夏大学动物科技学院/宁夏回族自治区反刍动物分子细胞育种重点实验室,银川750021 [2]宁夏新澳农牧有限公司,宁夏灵武750406

出　　处：《西南农业学报》2024年第1期210-222,共13页Southwest China Journal of Agricultural Sciences

基　　金：国家现代农业产业体系(CARS36);宁夏回族自治区重点研发项目(2021BEF01001、2021NXZD1、2022BBF02017)。

摘　　要：【目的】旨在探究前期GWAS筛选出的与荷斯坦奶牛产奶相关的二肽基肽酶样6(Dipeptidyl Peptidase⁃like 6,DPP6)和E3泛素连接酶基因(Parkin gene,PRKN)的生物学性质及表达特性,为后续验证DPP6和PRKN基因对于荷斯坦奶牛相关调控机制研究提供基础。【方法】利用ProtParam、Phyre2、NetPhos 3.1等软件分析DPP6和PRKN的理化性质与蛋白结构,并采用实时荧光定量PCR(Real⁃Time quantitative PCR,RT⁃qPCR)检测DPP6和PRKN基因在荷斯坦奶牛的心脏、脾脏、肝脏、肺脏、肾脏、乳腺、子宫、卵巢、瘤胃、黄体10个组织中的表达规律;通过脂多糖(Lipopolysaccharide,LPS)诱导乳腺上皮细胞(bMECs),初步检测DPP6和PRKN基因在炎症反应中的表达模式。【结果】DPP6蛋白编码863个氨基酸,与山羊的亲缘关系最近;PRKN蛋白编码488个氨基酸,其中甘氨酸含量最多,其序列与马鹿的相似性最高,保守型较强。本研究推测2个物种的DPP6和PRKN蛋白可能具有相似的生物学功能,并且2个蛋白均属于不稳定亲水性蛋白。DPP6和PRKN基因在组织中普遍表达。其中,DPP6基因在乳腺、肺脏、脾脏以及肾脏组织中的表达量显著高于其他组织,而PRKN基因在乳腺、肾脏、瘤胃组织中显著高表达。此外,相比于空白对照组,DPP6与PRKN基因均能够在LPS诱导的bMECs中极显著表达(DPP6基因极显著下调,PRKN基因极显著上调)。【结论】DPP6与PRKN基因可能在调控荷斯坦奶牛乳房炎症过程中发挥潜在作用,为后期荷斯坦奶牛DPP6与PRKN两个基因在奶牛bMECs炎症反应相关调控机制研究提供理论依据。【Objective】The purpose of the study was to explore the biological properties and tissue expression characteristics of Dipeptidyl Peptidase⁃like 6(DPP6)and E3 ubiquitin ligase gene(PRKN)genes that were screened by GWAS in the previous stage and related to milk production traits of Holstein.The study provided a basis for further verification of the regulatory mechanisms of DPP6 and PRKN genes on milk⁃production⁃related traits in Holstein.【Method】The physicochemical properties and protein structure of DPP6 and PRKN were analyzed by ProtParam,Phyre2,NetPhos 3.1 and other software,and Real⁃Time quantitative PCR was used.The expression patterns of DPP6 and PRKN in the heart,spleen,liver,lung,kidney,mammary gland,uterus,ovary,rumen and luteum of Holstein were detected by RT⁃qPCR;The relationship between DPP6 and PRKN genes and udder inflammation in Holstein dairy cows was examined by means of lipopo⁃lysaccharide(LPS)induced mammary epithelial cells(bMECs).The expression patterns of DPP6 and PRKN genes in inflammatory re⁃sponse were preliminarily detected.【Result】DPP6 protein encoded 863 amino acids and was closely related to goat.PRKN protein encoded 488 amino acids,among which glycine content was the highest,its sequence was the most similar to that of red deer,and the conserved type was strong.The study speculated that the DPP6 and PRKN proteins of two species may have similar biological functions,and both proteins were unstable hydrophilic proteins.DPP6 and PRKN genes were widely expressed in tissues.The expression of DPP6 in breast,lung,spleen and kidney tissues was significantly higher than that in other tissues,while the expression of PRKN was significantly higher in breast,kidney and rumen tissues.In addition,both DPP6 and PRKN genes were significantly expressed in LPS⁃induced bMECs compared with the blank control group(DPP6 gene was significantly down⁃regulated,and PRKN gene was highly upregulated).【Conclusion】DPP6 and PRKN genes may play a potential role in the regulation of ud

关 键 词：DPP6 PRKN 乳房炎症 荷斯坦奶牛 表达规律 

分 类 号：S823[农业科学—畜牧学]