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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p CAMBIA3301载体中,PCR和测序结果表明gma-miR1508a植物表达载体构建成功。
MicroRNAs are endogenous, non-coding small RNA molecules that play important regulatory roles in plant growth, development, and stress response. In this study, a target gene Glyma16g27800 of soybean gma-miR1508a was identified by 5’RLM-RACE method. In order to construct overexpression vector of gma-miR1508a, tobacco mosaic virus constitutive promoter 35S was used to control the overexpression of gma-miR1508a. A 101 bp precursor sequence with folded structure was amplified by PCR from soybean genomic DNA. The amplified fragment was subcloned into p CAMBIA3301 vector, PCR and sequencing results showed that gma-miR1508a plant expression vector was constructed successfully.