[1]倪志勇,于月华,任燕萍,等.大豆gma-miR1508a靶基因鉴定及植物表达载体构建[J].大豆科学,2015,34(06):1090-1092.[doi:10.11861/j.issn.1000-9841.2015.06.1090]
　NI Zhi-yong,YU Yue-hua,REN Yan-ping,et al.Validation of Selected gma-miR1508a Targets and Construction of Its Plant Expression Vectors[J].Soybean Science,2015,34(06):1090-1092.[doi:10.11861/j.issn.1000-9841.2015.06.1090]
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大豆gma-miR1508a靶基因鉴定及植物表达载体构建

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第34卷 期数: 2015年06期 页码: 1090-1092 栏目: 出版日期: 2015-12-25
Title:
Validation of Selected gma-miR1508a Targets and Construction of Its Plant Expression Vectors
作者:
倪志勇; 于月华; 任燕萍; 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;  REN Yan-ping;  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-miR1508a; 靶基因; 植物表达载体
Keywords:
Soybean;  gma-miR1508a;  Target gene;  Plant expression vector
DOI:
10.11861/j.issn.1000-9841.2015.06.1090
文献标志码:
A
摘要:
MicroRNAs是内源的非编码小RNA分子，在植物生长、发育和逆境响应过程中具有重要的调控功能。本研究采用5′ RLM-RACE方法鉴定了大豆gma-miR1508a的1个靶基因Glyma16g27800。为了构建gma-miR1508a的过表达载体，使用烟草花叶病毒组成型启动子35S控制gma-miR1508a的过表达，采用PCR方法从大豆基因组DNA中扩增了101 bp含有折回结构的前体序列，扩增片段被亚克隆至pCAMBIA3301载体中，PCR和测序结果表明gma-miR1508a植物表达载体构建成功。
Abstract:
MicroRNAs (miRNAs) are endogenous, noncoding, small RNAs that have essential regulatory functions in plant growth, development, and stress response processes.In this study, one target gene of gma-miR1508a, Glyma16g27800, was verified using a modified 5′ RLM-RACE (RNA ligasemediated rapid amplification of 5′ cDNA ends) assay. To generate an overexpression construct that constitutively overexpressed gma-miR1508a under the control of a cauliflower mosaic virus 35S promoter, a 101 bp fragment flanking the miRNA sequence including the fold-back structure was amplified from soybean genomic DNA by PCR. The amplified fragment was subcloned into the pCAMBIA3301 vector. PCR and sequencing results suggest that plant expression vector of gma-miR1508a gene was constructed successfully.This constructed vectors provided an effective too1 for the further study of gma-miR1508a gene function.

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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）；新疆维吾尔自治区自然科学基金(2014211A025，2013211B19)；新疆农业大学草业科学国家级重点学科。

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: 2016-01-07