CAJ | 학술논문

为了解开花麻竹(Dendrocalamus latiflorus)的DlAP2基因功能，采用RT-PCR和RACE技术克隆了miR172a靶基因AP2同源序列cDNA全长，命名为DlAP2。结果表明，DlAP2基因cDNA全长为1729 bp，包含5′端非编码区81 bp、开放阅读框1464 bp、3′端非编码区160 bp和24个碱基的Poly A尾巴，在编码框靠近3′端130 bp处有1个高度匹配miR172a的结合位点(CTGCAGCATCATCAGGATTCT)。DlAP2编码487个氨基酸的蛋白，具有两个AP2结构域，属于AP2/ERF家族AP2亚家族的AP2组，与来自其它单子叶植物的AP2蛋白均有较高同源性。RLM-5′ RACE分析表明，miR172a主要在靶序列的第11~12个碱基之间剪切靶基因DlAP2的miRNA。qRT-PCR结果表明，麻竹花芽中DlAP2基因的表达规律与miR172a表达变化正好相反，证明miR172a对DlAP2基因的表达具有调控作用。
위료해개화마죽(Dendrocalamus latiflorus)적DlAP2기인공능，채용RT-PCR화RACE기술극륭료miR172a파기인AP2동원서렬cDNA전장，명명위DlAP2。결과표명，DlAP2기인cDNA전장위1729 bp，포함5′단비편마구81 bp、개방열독광1464 bp、3′단비편마구160 bp화24개감기적Poly A미파，재편마광고근3′단130 bp처유1개고도필배miR172a적결합위점(CTGCAGCATCATCAGGATTCT)。DlAP2편마487개안기산적단백，구유량개AP2결구역，속우AP2/ERF가족AP2아가족적AP2조，여래자기타단자협식물적AP2단백균유교고동원성。RLM-5′ RACE분석표명，miR172a주요재파서렬적제11~12개감기지간전절파기인DlAP2적miRNA。qRT-PCR결과표명，마죽화아중DlAP2기인적표체규률여miR172a표체변화정호상반，증명miR172a대DlAP2기인적표체구유조공작용。
In order to understand the function ofDlAP2inDendrocalamus latiflorus, onemiR172a targeted gene named asDlAP2was cloned fromD. latiflorusby RT-PCR and RACE. Sequence analysis showed that the full length cDNA ofDlAP2was 1729 bp, including 5′ untranslated region (UTR) 81 bp, open reading frame (ORF) 1464 bp, 3′ UTR 351 bp, 24 bp polyA, and onemiR172a complementary site (CTGCAGCATCATCAGGATTCT) at 130 bp of the 3′ end in ORF.DlAP2encodes a putative protein with 487 amino acids with two AP2 domains, which indicate that it belongs to AP2 group of AP2 subfamily in AP2/ERF family. DlAP2 has a high homology with those AP2 from other monocots. RLM-5′ RACE analysis showed thatDlAP2was regulated bymiR172a through cutting mainly at the site between the 11th and 12th bases. Real-time quantitative PCR results showed that the expression pattern ofDlAP2was opposite to that ofmiR172a in lfower buds. These validated thatmiR172a played a regulatory role in regulating the expression ofDlAP2.