| 麻竹miR172a靶基因DlAP2的克隆及其表达 |
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| 引用本文： | 高志民,娄永峰,王丽丽,杨丽,赵韩生,陈东亮.麻竹miR172a靶基因DlAP2的克隆及其表达[J].热带亚热带植物学报,2015,23(3):245-251. |
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| 作者姓名： | 高志民 娄永峰 王丽丽 杨丽 赵韩生 陈东亮 |
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| 作者单位： | 国际竹藤中心, 竹藤科学与技术重点实验室, 北京 100102,国际竹藤中心, 竹藤科学与技术重点实验室, 北京 100102,国际竹藤中心, 竹藤科学与技术重点实验室, 北京 100102,国际竹藤中心, 竹藤科学与技术重点实验室, 北京 100102,国际竹藤中心, 竹藤科学与技术重点实验室, 北京 100102,1. 国际竹藤中心, 竹藤科学与技术重点实验室, 北京 100102;2. 北京市农林科学院生物中心, 北京 100097 |
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| 基金项目： | 国家林业局948 项目(2011-4-55)资助 |
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| 摘 要： | 为了解开花麻竹(Dendrocalamus latiflorus)的Dl AP2基因功能,采用RT-PCR和RACE技术克隆了mi R172a靶基因AP2同源序列c DNA全长,命名为Dl AP2。结果表明,Dl AP2基因c DNA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mi R172a的结合位点(CTGCAGCATCATCAGGATTCT)。Dl AP2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,mi R172a主要在靶序列的第11~12个碱基之间剪切靶基因Dl AP2的mi RNA。q RT-PCR结果表明,麻竹花芽中Dl AP2基因的表达规律与mi R172a表达变化正好相反,证明mi R172a对Dl AP2基因的表达具有调控作用。
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| 关 键 词： | 麻竹 AP2基因 miR172a 基因表达 |
| 收稿时间： | 2014/9/18 0:00:00 |
| 修稿时间： | 2014/11/14 0:00:00 |
Cloning and Expression Analysis of miR172a Targeted Gene DlAP2 in Dendrocalamus latiflorus |
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| GAO Zhi-min,LOU Yong-feng,WANG Li-li,YANG Li,ZHAO Han-sheng and CHEN Dong-liang.Cloning and Expression Analysis of miR172a Targeted Gene DlAP2 in Dendrocalamus latiflorus[J].Journal of Tropical and Subtropical Botany,2015,23(3):245-251. |
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| Authors: | GAO Zhi-min LOU Yong-feng WANG Li-li YANG Li ZHAO Han-sheng and CHEN Dong-liang |
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| Institution: | International Center for Bamboo and Rattan, Key Laboratory on Science and Technology of Bamboo and Rattan, Beijing 100102, China,International Center for Bamboo and Rattan, Key Laboratory on Science and Technology of Bamboo and Rattan, Beijing 100102, China,International Center for Bamboo and Rattan, Key Laboratory on Science and Technology of Bamboo and Rattan, Beijing 100102, China,International Center for Bamboo and Rattan, Key Laboratory on Science and Technology of Bamboo and Rattan, Beijing 100102, China,International Center for Bamboo and Rattan, Key Laboratory on Science and Technology of Bamboo and Rattan, Beijing 100102, China and 1. International Center for Bamboo and Rattan, Key Laboratory on Science and Technology of Bamboo and Rattan, Beijing 100102, China;2. Beijing Agro-Biotechnology Research Center, Beijing 100097, China |
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| Abstract: | In order to understand the function of DlAP2 in Dendrocalamus latiflorus, one miR172a targeted gene named as DlAP2 was cloned from D. latiflorus by RT-PCR and RACE. Sequence analysis showed that the full length cDNA of DlAP2 was 1729 bp, including 5' untranslated region (UTR) 81 bp, open reading frame (ORF) 1464 bp, 3' UTR 351 bp, 24 bp polyA, and one miR172a complementary site (CTGCAGCATCATCAGGATTCT) at 130 bp of the 3' end in ORF. DlAP2 encodes 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 that DlAP2 was regulated by miR172a through cutting mainly at the site between the 11th and 12th bases. Real-time quantitative PCR results showed that the expression pattern of DlAP2 was opposite to that of miR172a in flower buds. These validated that miR172a played a regulatory role in regulating the expression of DlAP2. |
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| Keywords: | Dendrocalamus latiflorus AP2 gene miR172a Gene expression |
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