| 猪LPAR3基因克隆及其在子宫内膜细胞的表达 |
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| 引用本文： | 廖妙容,吴龙,张爱玲,钟玉宜,温丽娟,张豪,袁晓龙,李加琪,张哲.猪LPAR3基因克隆及其在子宫内膜细胞的表达[J].华南农业大学学报,2019,40(2):31-39. |
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| 作者姓名： | 廖妙容 吴龙 张爱玲 钟玉宜 温丽娟 张豪 袁晓龙 李加琪 张哲 |
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| 作者单位： | 广东省农业基因组学与分子育种重点实验室/华南农业大学 动物科学学院,广东 广州,510642;广东第二师范学院 生物与食品工程学院/广东高校应用生态工程技术开发中心,广东 广州,510303 |
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| 基金项目： | 科技部科技基础工作专项（2014FY120800）；国家现代农业产业技术体系项目（CARS-35）；广东扬帆计划（2014YT02H042）；广东省科技计划项目（2017A020208043） |
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| 摘 要： | 【目的】获得猪LPAR3基因完整mRNA及基因结构,研究其启动子活性;探究LPAR3基因在子宫内膜的转录调控及可能影响母猪产仔的机制。【方法】应用5'RACE和3'RACE技术获取LPAR3基因完整mRNA序列;预测5'调控区潜在的启动子转录因子结合位点及CpG岛,构建不同长度的启动子双荧光素酶报告基因重组载体,与pRL-TK质粒共同转染至猪子宫内膜细胞,检测启动子活性;应用RT-qPCR比较LPAR3基因在妊娠第12天的二花脸猪和长大二元猪子宫内膜的相对表达量;应用亚硫酸氢钠修饰后测序比较LPAR3基因在妊娠第12天的二花脸猪和长大二元猪子宫内膜的甲基化状态。【结果】猪LPAR3 mRNA全长为2 127 bp,其中5'UTR和3'UTR的长度分别为202和860 bp,CDS区为1 065 bp。克隆获得包括LPAR3转录起始位点上游3 080 bp (–2 430/+650bp)的5'调控序列,分析预测显示该调控区不存在TATA box,存在GC元件、CPBP及糖皮质激素受体IR3等调控因子结合位点,且在–190/–84和–44/+651 bp处存在2个潜在CpG岛。成功构建9个不同长度的5'缺失报告重组载体并转染猪子宫内膜细胞。双荧光素酶活性检测结果显示,启动子P4(+454/+80 bp)的转录活性最高,其次是P6(–123/+80 bp)。RT-qPCR结果显示,妊娠第12天二花脸猪LPAR3基因在子宫内膜的表达量高于在其他组织的表达量,且极显著高于在妊娠第12天长大二元猪子宫内膜的表达量,LPAR3在2个猪种子宫内膜均处于低甲基化状态且差异不显著。【结论】猪LPAR3 mRNA全长为2 127 bp,妊娠第12天LAPR3基因在二花脸猪子宫内膜表达高于其在长大二元猪子宫内膜的表达,显示LPAR3可能参与了猪早期妊娠并影响产仔数。
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| 关 键 词： | 猪 LPAR3基因 启动子 子宫内膜 妊娠 甲基化 |
| 收稿时间： | 2018/5/19 0:00:00 |
Cloning of porcine LPAR3 gene and its expression in endometrial cells |
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| LIAO Miaorong,WU Long,ZHANG Ailing,ZHONG Yuyi,WEN Lijuan,ZHANG Hao,YUAN Xiaolong,LI Jiaqi and ZHANG Zhe.Cloning of porcine LPAR3 gene and its expression in endometrial cells[J].Journal of South China Agricultural University,2019,40(2):31-39. |
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| Authors: | LIAO Miaorong WU Long ZHANG Ailing ZHONG Yuyi WEN Lijuan ZHANG Hao YUAN Xiaolong LI Jiaqi and ZHANG Zhe |
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| Institution: | Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/College of Animal Science, South China Agricultural University, Guangzhou 510642, China,Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/College of Animal Science, South China Agricultural University, Guangzhou 510642, China,Biology and Food Engineering Institute, Guangdong University of Education/Guangdong Development Center of Applied Ecology and Ecological Engineering in Universities, Guangzhou 510303, China,Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/College of Animal Science, South China Agricultural University, Guangzhou 510642, China,Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/College of Animal Science, South China Agricultural University, Guangzhou 510642, China,Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/College of Animal Science, South China Agricultural University, Guangzhou 510642, China,Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/College of Animal Science, South China Agricultural University, Guangzhou 510642, China,Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/College of Animal Science, South China Agricultural University, Guangzhou 510642, China and Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/College of Animal Science, South China Agricultural University, Guangzhou 510642, China |
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| Abstract: | |
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| Keywords: | pig LPAR3 gene promoter endometrium gestation methylation |
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