猪Neuronatin基因在pTr2细胞内葡萄糖转运中的功能研究  

Study on the Effect of Porcine Neuronatin Gene on Glucose Transport in pTr2 Cells

作　　者：邢萍萍 严冠豪 谭宝华 乔佳鑫 王珊珊 洪林君 郑恩琴[1] 黄思秀[1] 顾婷[1] XING Pingping;YAN Guanhao;TAN Baohua;QIAO Jiaxin;WANG Shanshan;HONG Linjun;ZHENG Enqin;HUANG Sixiu;GU Ting(National Engineering Technology Research Center for Pig Seed Industry&College of Animal Science and Technology,South China Agricultural University,Guangdong Guangzhou 510642,China)

机构地区：[1]国家生猪种业工程技术研究中心&华南农业大学动物科技学院,广东广州510642

出　　处：《中国畜牧杂志》2022年第1期102-108,共7页Chinese Journal of Animal Science

基　　金：广东省自然科学基金(2019B1515210014);国家自然科学基金(31802036);广东省重点领域研究与发展计划(2018B020203002)。

摘　　要：本研究旨在探索印记基因NNAT 2个转录本NNAT-α、NNAT-β在猪胎盘滋养层细胞(pTr2)中的功能。实验通过NCBI数据库公布的猪NNAT-α、NNAT-β的CDs序列信息设计引物,并构建了pcDNA3.1-NNAT-α和pcDNA3.1-NNAT-β真核表达载体,经过双酶切、连接转化、测序鉴定后,将表达载体转染至pTr2细胞,48 h后通过酶标仪检测细胞内葡萄糖含量的变化,RT-PCR检测NNAT基因、葡萄糖转运基因(GLUT1、GLUT3)和PI3K-AKT通路基因的表达。结果表明在pTr2细胞中过表达NNAT-α、NNAT-β可显著提高细胞内葡萄糖含量、葡萄糖转运基因(GLUT1、GLUT3)和PI3K-AKT通路基因mRNA的表达量。This experiment was conducted to explore the functions of the two transcripts of the imprinted gene NNAT,NNAT-αand NNAT-β,in porcine placental trophoblast cells(pTr2).In this study,primers were designed based on the CDs sequence of pig NNAT-αand NNAT-βin the NCBI database,and the expression vectors of pcDNA3.1-NNAT-αand pcDNA3.1-NNAT-βwere constructed.After double enzyme digestion and sequencing identification,they were transfected into the pTr2 cells.After transfection for 48 hours,the changes in glucose contents in the cells were detected by a microplate reader,and the expression levels of NNAT gene,glucose transport genes(GLUT1,GLUT3)and PI3K-AKT pathway genes were detected by RT-PCR.The results showed that overexpression of NNAT-αand NNAT-βin pTr2 cells can significantly increase the intracellular glucose content,glucose transport genes(GLUT1,GLUT3)and the genes in the PI3K-AKT pathway.

关 键 词：印记基因 NNAT 真核表达载体 葡萄糖转运 

分 类 号：S828.2[农业科学—畜牧学]