[1]王欣,范洋洋,陈晨,等.家蚕ｂｍｏ-ｍｉＲ-３４对ＢｍＥ７４基因表达的调控[J].江苏农业学报,2016,(01):51-57.[doi:10.3969/j.issn.1000-4440.2016.01.008]
　WANG Xin,FAN Yang-yang,CHEN Chen,et al.Expression regulation of BmE74 gene by bmo-miR-34 in Bombyx mori[J].,2016,(01):51-57.[doi:10.3969/j.issn.1000-4440.2016.01.008]
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家蚕ｂｍｏ-ｍｉＲ-３４对ＢｍＥ７４基因表达的调控()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年01期
页码:
51-57
栏目:
遗传育种·生理生化
出版日期:
2016-01-08

文章信息/Info

Title:
Expression regulation of BmE74 gene by bmo-miR-34 in Bombyx mori
作者:
王欣12; 范洋洋12; 陈晨12; 谢雨辰12; 蒋涛12; 汪生鹏12; 沈兴家12
（1.江苏科技大学生物技术学院江苏省蚕桑生物学与生物技术重点实验室，江苏镇江212018；2.中国农业科学院蚕业研究所，江苏镇江212018）
Author(s):
WANG Xin12; FAN Yang-yang12; CHEN Chen12; XIE Yu-chen12; JIANG Tao12; WANG Sheng-peng12; SHEN Xing-jia12
(1.Jiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212018, China;2.Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212018, China)
关键词:
miR-34; 家蚕; BmE74基因; 表达调控
Keywords:
miR-34; Bombyx mori; BmE74 gene; expression regulation
分类号:
S881.2;Q78
DOI:
10.3969/j.issn.1000-4440.2016.01.008
文献标志码:
A
摘要:
为了研究家蚕bmo-miR-34（一类高度保守的miRNA）对E74基因（BmE74）的表达调控作用，首先利用生物信息学软件RNAhybrid和RNA22进行结合位点预测，然后从家蚕丝腺组织中克隆了bmo-miR-34前体序列，分别构建bmo-miR-34表达载体和靶基因BmE74 3′UTR重组表达载体，利用双报告基因荧光检测系统在细胞水平研究bmo-miR-34对BmE74基因的转录后调控作用。生物信息学预测结果表明，BmE74 3′UTR具有bmo-miR-34潜在结合位点；体外转染试验结果显示，与对照相比，转染bmo-miR-34重组质粒的细胞荧光素酶活性极显著下调（P<0.01），但是当加入人工合成的bmo-miR-34抑制物后荧光素酶活性显著上调（P<0.01）。说明bmo-miR-34对BmE74的转录后表达具有抑制作用。
Abstract:
In order to study the expression regulation of bmo-miR-34 (a highly conserved miRNA) on BmE74 gene in Bombyx mori, bioinformatics softwares RNAhybrid and RNA22 were used to identify the binding site in BmE74 3′UTR. Precursor bmo-miR-34 sequence was cloned afterwards, and bmo-miR-34 recombinant expression vector and E74 3′UTR expression vector were constructed for transient expression assay. Dual-luciferase reporter gene system was applied to detect the role of bmo-miR-34 in the posttranscriptional regulation of BmE74 gene at the cellular level. The predicted results of bioinformatics showed that there were bmo-mir-34 potential binding sites on E74 3′UTR. The results of transfection experiments in vitro revealed that compared with the control group the luciferase activity was significantly decreased in the BmN cells co-transfected with bmo-miR-34 recombinant plasmid （P<0.01）, and ascended distinctly after transfected with synthetic bmo-miR-34 inhibitor （P<0.01）. It is indicated that bmo-miR-34 inhibits E74 gene posttranscriptional expression.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期：2015-04-09基金项目：国家自然科学基金项目(31172266/C1703)；江苏省普通高校研究生科研创新计划项目(CXZZ12_0726) 作者简介：王欣(1984-)，女，河北唐山人，博士研究生，主要从事家蚕分子生物学研究。 （Tel）0511-85601052；（E-mail）wang7xin23@yeah.net通讯作者：沈兴家，（Tel）0511-85601052；（E-mail）shenxjsri@163.com
更新日期/Last Update: 2017-05-08