[1]倪志勇,于月华,陈全家,等.大豆gma-miR1514b生物信息学分析及人工microRNA植物表达载体构建[J].大豆科学,2016,35(01):46-51.[doi:10.11861/j.issn.1000-9841.2016.01.0046]
　NI Zhi-yong,YU Yue-hua,CHEN Quan-jia,et al.Bioinformatic Analysis of gma-miR1514b and Construction of Its Artificial miRNA Plant Expression Vectors[J].Soybean Science,2016,35(01):46-51.[doi:10.11861/j.issn.1000-9841.2016.01.0046]
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大豆gma-miR1514b生物信息学分析及人工microRNA植物表达载体构建

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年01期 页码: 46-51 栏目: 出版日期: 2016-01-20
Title:
Bioinformatic Analysis of gma-miR1514b and Construction of Its Artificial miRNA Plant Expression Vectors
作者:
倪志勇; 于月华; 陈全家; 曲延英
新疆农业大学 农学院/新疆农业大学农业生物技术重点实验室, 新疆 乌鲁木齐 830052
Author(s):
NI Zhi-yong;  YU Yue-hua;  CHEN Quan-jia;  QU Yan-ying
College of Agronomy, Xinjiang Agricultural University / Key Laboratory of Agricultural Biological Technology, Xinjiang Agricultural University, Urumqi 830052, China
关键词:
大豆; gma-miR1514b; amiRNA; 启动子
Keywords:
Soybean;  gma-miR1514b;  amiRNA;  Promoter
DOI:
10.11861/j.issn.1000-9841.2016.01.0046
文献标志码:
A
摘要:
MicroRNAs在植物发育与逆境响应等多个方面具有重要的调节作用。本研究采用生物信息学方法分析gma-miR1514b的靶基因和启动子中的顺式作用元件。PLACE和PlantCARE启动子在线预测工具分析表明gma-miR1514b启动子序列具有TATA-box、CAAT-box 基本的顺式作用元件和一些参与光、逆境和植物激素应答相关的顺式作用元件。PMRD软件预测获得5个gma-miR1514b靶基因，分别与拟南芥CA2、NTL9和ANAC014基因同源。通过重叠PCR方法替换拟南芥At-MIR319的成熟链和互补链序列，构建含有amiR1514b前体的植物表达载体amiR1514b-pCAMBIA3301。
Abstract:
MicroRNAs play fundamental regulatory roles in various aspects of plant development and adaptive responses to stresses. In this study，the target gene and the promoter of gma-miR1514b gene were predicted and analyzed by bioinformatics methodology PMRD database analysis showed that precursor sequence length of gma-miR1514b was 109 bp, located in soybean 1 497 668-1 497 776［+］of chromosome 17, and its mature sequence length was 21 bp. The predicted transcription start site of gma-miR1514b was found within 762 bp upstream of the precursor sequence.The database of PLACE and PlantCARE analysis showed that the promoter sequence contains basic elements TATA-box, CAAT-box and some cis-acting elements involved in response to light, stresses and plant hormones PMRD software analysis showed that 5 transcripts were targeted by gma-miR1514b, these targets were homologous with CA2, NTL9 and ANAC014 genes in Arabidopsis, respectively. The amiR1514b sequence was engineered into the miRNA and miRNA* of At-MIR319 using overlapping PCR and the amiR1514b backbone was transferred into the plant expression vector pCAMBIA3301.

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备注/Memo

基金项目：中国博士后科学基金（2015M582741）；国家自然科学基金（31360264）；新疆维吾尔自治区高校科研计划科学研究重点项目（XJEDU2014I015）；新疆农业大学校前期资助课题（XJAU201313）；新疆维吾尔自治区优秀博士后普通资助。

第一作者简介：倪志勇（1981-），男，博士，副教授，主要从事植物逆境分子生物学研究。E-mail：nizhiyong@126.com。

更新日期/Last Update: 2016-01-28