miR-122靶向调控鸡BZW2基因的表达

发布时间：2018-01-13 17:42

  本文关键词：miR-122靶向调控鸡BZW2基因的表达　出处：《农业生物技术学报》2016年11期 　论文类型：期刊论文

  更多相关文章： 鸡 miR- 肝脏 BZW基因

【摘要】：miR-122(Micro RNA-122)是在肝脏中高表达的mi RNA,在肝脏的生长发育和脂类代谢等过程发挥重要作用,BZW2(basic leucine zipper and W2 domains 2)主要参与蛋白质合成代谢,本研究目的是确定BZW2是否为mi R-122调控的靶基因。通过生物信息学预测mi R-122的靶基因,并分析BZW2的3-UTR区域与mi R-122种子区的配对情况,再用双荧光素酶报告系统和突变实验证明mi R-122作用于BZW2的3'UTR。用荧光定量PCR检测转染mi RNA-122mimic的鸡(Gallus gallus)肝癌细胞系(leghorn hepatocellar,LMH)细胞和转染LNA-antimi R-122的鸡原代肝细胞中BZW2的表达。鸡BZW2的3'-UTR区域与mi R-122种子区互补配对,荧光素酶报告基因和突变实验分析表明,mi R-122能够通过与BZW2 3'-UTR区结合抑制基因的表达;将mi R-122在LMH细胞中过表达后,发现BZW2 m RNA表达水平显著下降;利用LNAantimi R-122抑制鸡肝脏原代细胞中的mi R-122后,BZW2 m RNA表达水平呈显著性上升。结果证明BZW2是mi R-122的靶基因,其在m RNA水平上受到mi R-122的负性调控,本研究为揭示mi R-122在鸡肝脏中的广泛的功能提供了理论依据。
[Abstract]:RNA-122) is a highly expressed miR-122(Micro in the liver, which plays an important role in the growth and development of the liver and lipid metabolism. BZW2(basic leucine zipper and W2 domains 2) is mainly involved in protein synthesis and metabolism. The aim of this study was to determine whether BZW2 was the target gene regulated by miR-122 and to predict the target gene of miR-122 by bioinformatics. The pairing of 3-UTR region of BZW2 with the seed region of mi R-122 was analyzed. Double luciferase reporting system and mutagenesis test proved that mi R-122 acted on 3UTR of BZW2. The chicken transfected with mi RNA-122mimic was detected by fluorescence quantitative PCR. Gallus gallus) hepatoma cell line Leghorn hepatocellar. LMH). The expression of BZW2 in cells and primary hepatocytes transfected with LNA-antimi R-122. The 3H-UTR region of chicken BZW2 was matched with the seed region of miR-122. Luciferase reporter gene and mutagenesis analysis showed that mil R-122 could inhibit the gene expression by binding to BZW2 3 + -UTR region. After overexpression of miR-122 in LMH cells, it was found that the expression of BZW2 m RNA decreased significantly. LNAantimi R-122 was used to inhibit mi R-122 in primary chicken liver cells. The expression of BZW2 m RNA was significantly increased. The results showed that BZW2 was the target gene of miR-122, which was negatively regulated by miR-122 at m RNA level. This study provides a theoretical basis for revealing the wide range of functions of mi-122 in chicken liver.
【作者单位】： 常熟理工学院生物与食品工程学院;江苏省家禽科学研究所;南京医科大学附属常州第二人民医院肿瘤研究所;
【基金】：国家自然科学基金项目(No.31272438和No.31472091) 江苏省自然科学基金(No.BK20151257) 苏州市科技计划项目(No.SYN201516)
【分类号】：S831.2
【正文快照】： 究所,常州213003Micro RNA(mi RNA)是长度约22个核苷酸的一类非编码单链RNA分子,其在生命体的各个生物过程中发挥着重要的作用(Bartel,2004;Lee etal.,1993)。mi RNA主要通过与靶基因m RNA的3'-UTR区互补配对来调节靶基因的表达(Doench,Sharp,2004)。mi R-122是肝脏中表达量最，

本文编号：1419891