| 花生AhDGAT2a基因启动子的克隆和功能验证 |
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| 引用本文： | 郑玲,史灵敏,田海莹,单雷,边斐,郭峰,宣宁,万书波,彭振英.花生AhDGAT2a基因启动子的克隆和功能验证[J].作物学报,2016,42(7):1094-1099. |
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| 作者姓名： | 郑玲 史灵敏 田海莹 单雷 边斐 郭峰 宣宁 万书波 彭振英 |
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| 作者单位： | 1山东省农业科学院生物技术研究中心 / 山东省作物遗传改良与生态生理重点实验室，山东济南 250100； 2山东大学生命科学学院，山东济南 250100；3新疆农业大学农学院，新疆乌鲁木齐 830000 |
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| 基金项目： | 本研究由山东省自然科学基金项目(ZR2013CM036)和山东省良种工程项目(2014-2017)的资助。 |
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| 摘 要： | 二酰甘油酰基转移酶(DGAT)是三酰甘油(TAG)合成途径的限速酶,对脂肪酸合成的调节具有关键作用。为了研究AhDGAT2a的表达调控,利用Genome Walking方法从鲁花14基因组中克隆了AhDGAT2a上游5￠侧翼调控区1200 bp序列,即AhDGAT2a启动子(pAhDGAT2a)序列,并利用生物信息学软件分析其包含的调控元件,发现其含有多个TATA-box和CAAT-box、光调控元件、胁迫防御相关元件和激素响应元件。用pAhDGAT2a构建pAhDGAT2a:GUS植物表达载体并转化烟草品种SR1。利用组织染色法鉴定转基因烟草的GUS表达模式,发现在转基因烟草的各个器官均有GUS酶活,在柱头、花药和幼嫩种子中表达量较高,说明pAhDGAT2a具有一定的组成型启动子活性。
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| 关 键 词： | 花生 DGAT2a基因 启动子 功能验证 |
| 收稿时间： | 2016-01-11 |
Cloning and Functional Analysis of Peanut AhDGAT2a Promoter? |
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| ZHENG Ling,SHI Ling-Min,TIAN Hai-Ying,SHAN Lei,BIAN Fei,GUO Feng.Cloning and Functional Analysis of Peanut AhDGAT2a Promoter?[J].Acta Agronomica Sinica,2016,42(7):1094-1099. |
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| Authors: | ZHENG Ling SHI Ling-Min TIAN Hai-Ying SHAN Lei BIAN Fei GUO Feng |
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| Institution: | 1.Biotechnology Research Center, Shandong Academy of Agricultural Sciences / Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Jinan 250100, China;2.College of Life Science, Shandong University, Jinan 250100, China;3.College of Agronomy, Xinjiang Agricultural University, Urumqi 830000, China |
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| Abstract: | Diacylglycerol acyltransferase (DGAT) is a rate-limiting enzyme in triacylglycerol (TAG) biosynthesis pathway. In this study, GenomeWalking method was used for cloning the promoter sequence of AhDGAT2a gene from Luhua 14, and finally a 1200 bp fragment flanking 5′-upstream of AhDGAT2a was obtained and named as pAhDGAT2a. The crucial regulatory elements in pAhDGAT2a were further analyzed with software PlantCARE. There were many TATA-box, CAAT-box, light regulation, stress and defense response and hormone response elements. To assess the activity of pAhDGAT2a, we constructed pAhDGAT2a:GUS cassettes and introduced it into the tobacco SR1 genome by Agrobacterium-mediated transformation. Expression pattern was monitored by histochemical staining. Results showed that GUS activity driven by the pAhDGAT2a was detected in almost all vegetative and reproductive tissues, with a higher expression level in stigma, anther and young seeds than in the other organs, indicating thatpAhDGAT2a has a constitutive promoter activity. |
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| Keywords: | Arachis hypogaea L DGAT2a gene Promoter Function analysis |
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