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2025年4月5日 星期六
  2016, Vol. 24 Issue (11): 1734-1740    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
miR-122靶向调控鸡BZW2基因的表达
李艳艳1,王星果2,郁建锋1,邵芳3,3,张燕萍3,3,顾志良1
1. 常熟理工学院
2. 江苏省家禽科学研究所
3.
miR-122 Targets to Regulate BZW2 Gene Expression in Chicken(Gallus gallus)
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摘要 miR-122(MicroRNA-122)是在肝脏中高表达的miRNA,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miR-122调控的靶基因。通过生物信息学预测miR-122的靶基因,并分析BZW2的3-UTR区域与miR-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miR-122作用于BZW2的3'UTR。用荧光定量PCR检测转染miRNA-122mimic的鸡(Gallus gallus)肝癌细胞系(leghorn hepatocellar, LMH)细胞和转染LNA-antimiR-122的鸡原代肝细胞中BZW2的表达。鸡BZW2的3'-UTR区域与miR-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,miR-122能够通过与BZW2 3'-UTR区结合抑制基因的表达;将miR-122在LMH细胞中过表达后,发现BZW2 mRNA表达水平显著下降;利用LNA-antimiR-122抑制鸡肝脏原代细胞中的miR-122后,BZW2 mRNA表达水平呈显著性上升。结果证明BZW2是miR-122的靶基因,其在mRNA水平上受到miR-122的负性调控,本研究为揭示miR-122在鸡肝脏中的广泛的功能提供了理论依据。
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李艳艳
王星果
郁建锋
邵芳
张燕萍
顾志良
关键词 miR-122肝脏BZW2基因    
Abstract:MicroRNAs are a class of tiny, single-stranded noncoding RNAs that can target mRNA via base pairing and lead to translational repression or mRNA degradation at the level of gene posttranscription, which are involved in organ development, cell differentiation, proliferation, apoptosis and other biological processes, closely related with many diseases. Basic leucine zipper and W2 domains 2 (BZW2) functions in protein synthesis metabolism. The purpose of this study was to validate whether miR-122 could inhibit BZW2 gene expression in chicken (Gallus gallus). The bioinformatics method was used to investigate whether miR-122 could pair chicken BZW2 3'-UTR, furthermore dual-luciferase report system and mutant assay were used to validate that miR-122 repressed BZW2 expression. qRT-PCR was used to detect the expression level of BZW2 in leghorn hepatocellar (LHM) cells transfected with miRNA-122mimic and primary chicken hepatocytes transfected with LNA-antimiR-122. Bioinformatics prediction and dual luciferase report system proved that BZW2 was target gene of miR-122, target site mutation assay showed the complementary site of 3'UTR with miR-122 seeds region were target site of miR-122. After overexpression of miR-122 in leghorn hepatocellar (LMH) cells, BZW2 mRNA expression level decreased obviously. Using LNA-antimiR-122 to knockdown the miR-122 expression in primary chicken hepatocytes, the target gene BZW2 mRNA expression level increased significantly. These data suggested that BZW2 gene was target gene of miR-122 and BZW2 mRNA was negatively regulated by miR-122 in chicken hepatocytes. It will provide scientific basis on revealing a wide range of functions in the liver of miR-122 in chicken.
Key wordsChicken    miR-122    Liver    Basic leucine zipper and W2 domains 2(BZW2) gene
收稿日期: 2016-07-12      出版日期: 2016-10-01
基金资助:国家自然科学基金项目
通讯作者: 李艳艳     E-mail: 469321054@qq.com
引用本文:   
李艳艳 王星果 郁建锋 邵芳 张燕萍 顾志良. miR-122靶向调控鸡BZW2基因的表达[J]. , 2016, 24(11): 1734-1740.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2016/V24/I11/1734
 
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