本文關(guān)鍵詞：miR-122靶向調(diào)控雞BZW2基因的表達　出處：《農(nóng)業(yè)生物技術(shù)學報》2016年11期 　論文類型：期刊論文

  更多相關(guān)文章： 雞 miR- 肝臟 BZW基因

【摘要】：miR-122(Micro RNA-122)是在肝臟中高表達的mi RNA,在肝臟的生長發(fā)育和脂類代謝等過程發(fā)揮重要作用,BZW2(basic leucine zipper and W2 domains 2)主要參與蛋白質(zhì)合成代謝,本研究目的是確定BZW2是否為mi R-122調(diào)控的靶基因。通過生物信息學預測mi R-122的靶基因,并分析BZW2的3-UTR區(qū)域與mi R-122種子區(qū)的配對情況,再用雙熒光素酶報告系統(tǒng)和突變實驗證明mi R-122作用于BZW2的3'UTR。用熒光定量PCR檢測轉(zhuǎn)染mi RNA-122mimic的雞(Gallus gallus)肝癌細胞系(leghorn hepatocellar,LMH)細胞和轉(zhuǎn)染LNA-antimi R-122的雞原代肝細胞中BZW2的表達。雞BZW2的3'-UTR區(qū)域與mi R-122種子區(qū)互補配對,熒光素酶報告基因和突變實驗分析表明,mi R-122能夠通過與BZW2 3'-UTR區(qū)結(jié)合抑制基因的表達;將mi R-122在LMH細胞中過表達后,發(fā)現(xiàn)BZW2 m RNA表達水平顯著下降;利用LNAantimi R-122抑制雞肝臟原代細胞中的mi R-122后,BZW2 m RNA表達水平呈顯著性上升。結(jié)果證明BZW2是mi R-122的靶基因,其在m RNA水平上受到mi R-122的負性調(diào)控,本研究為揭示mi R-122在雞肝臟中的廣泛的功能提供了理論依據(jù)。
[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.
【作者單位】： 常熟理工學院生物與食品工程學院;江蘇省家禽科學研究所;南京醫(yī)科大學附屬常州第二人民醫(yī)院腫瘤研究所;
【基金】：國家自然科學基金項目(No.31272438和No.31472091) 江蘇省自然科學基金(No.BK20151257) 蘇州市科技計劃項目(No.SYN201516)
【分類號】：S831.2
【正文快照】： 究所,常州213003Micro RNA(mi RNA)是長度約22個核苷酸的一類非編碼單鏈RNA分子,其在生命體的各個生物過程中發(fā)揮著重要的作用(Bartel,2004;Lee etal.,1993)。mi RNA主要通過與靶基因m RNA的3'-UTR區(qū)互補配對來調(diào)節(jié)靶基因的表達(Doench,Sharp,2004)。mi R-122是肝臟中表達量最，

本文編號：1419891