NI Zhi-yong,YU Yue-hua,CHEN Quan-jia,et al.Bioinformatic Analysis of gma-miR1510a and Construction of Its Artificial MiRNA Plant Expression Vectors[J].Soybean Science,2016,35(02):239-244.[doi:10.11861/j.issn.1000-9841.2016.02.0239]
大豆gma-miR1510a生物信息学分析及人工microRNA植物表达载体构建
- Title:
- Bioinformatic Analysis of gma-miR1510a and Construction of Its Artificial MiRNA Plant Expression Vectors
- 关键词:
- 大豆; gma-miR1510a; amiRNA; 启动子
- Keywords:
- Soybean; gma-miR1510a; amiRNA; Promoter
- 文献标志码:
- A
- 摘要:
- MicroRNAs (miRNAs)是最近发现的一类广泛存在的内生的小RNAs,在转录水平负调控基因的表达。本研究采用生物信息学方法分析gma-miR1510a的靶基因和启动子中的顺式作用元件。PLACE和PlantCARE启动子数据库分析表明gma-miR1510a启动子具有一些参与非生物胁迫、植物激素、组织特异表达和光应答元件。PMRD软件预测获得9个gma-miR1510a靶基因,属于抗病蛋白和ATP结合家族成员。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amiR1510a前体的植物表达载体amiR1510a-pCAMBIA3301。
- Abstract:
- Abstract:MicroRNAs (miRNAs) are newly discovered extensive endogenous small RNAs, which negatively regulate gene expression at the post-transcription levels.In this study,the target gene and the promoter of gmamiR1510a gene were predicted and analyzed by bioinformatics methodology.Many response elements for abiotic stress, plant hormones, tissue-specific expression and light were predicted by the database search programs PLACE and PlantCARE- PMRD software analysis showed that 9 transcripts were targeted by gma-miR1510a, these targets belonged to disease resistance protein (TIR-NBS-LRR class) and ATP binding.The amiR1510a sequence was engineered into the miRNA and miRNA* of At-MIR319 using overlapping PCR and the amiR1510a backbone was transferred into the plant expression vector pCAMBIA3301.
参考文献/References:
[1]Zhang B H, Pan X P, Cobb G P, et al. Plant microRNA: A small regulatory molecule with big impact[J].Developmental Biology, 2006, 289:3-16.
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备注/Memo
基金项目:中国博士后科学基金资助项目(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);新疆维吾尔自治区优秀博士后普通资助。