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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (10): 146-150.doi: 10.11924/j.issn.1000-6850.2011-3324

所属专题： 生物技术； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

苹果赤霉素3-氧化酶1(MdGA3ox1)启动子区

赵慧君 仝宗永 余海忠 孙永林   

  • 收稿日期:2011-11-09 修回日期:2012-01-21 出版日期:2012-04-05 发布日期:2012-04-05

Cloning and Sequence Analysis of Promoter Region of Apple (malus x demestic) Gibberellin 3-oxidase1

  • Received:2011-11-09 Revised:2012-01-21 Online:2012-04-05 Published:2012-04-05

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摘要：

通过研究苹果赤霉素3-氧化酶1(MdGA3ox1)的启动子来探索MdGA3ox1的转录调控机制。采用染色体步移法结合锚定PCR法扩增了MdGA3ox1的上游序列，用PLANT CARE启动子分析软件分析此启动子序列，并把其构建到载体上，转化入根癌农杆菌中初步验证了其功能。获得一个754 bp的DNA序列，分析此序列发现有CAATbox、TATAbox、赤霉素、光、乙烯、脱落酸等顺式反应元件。把此序列构建到pCAMBIA 1381载体上，并把此质粒命名为p1381-GA3ox1p。把p1381-GA3ox1p转化入农杆菌EHA105中，利用GUS染色液染色，农杆菌呈蓝色。获得了MdGA3ox1的上游序列，此序列上含有与GA调节相关的顺式反应元件，并且初步证明此序列具有启动子活性。

关键词: 电阻值, 电阻值

Abstract:

The aim was to study the transcription regulation mechanism of MdGA3ox1 gene by studying the promoter region of MdGA3ox1 gene. The upstream sequence of ‘Fuji’ (Malus x domestica Borkh. cv. Fuji) MdGA3ox1 was attained by Genome Walking binding anchored PCR. The functional elements were analyzed by PLANTCARE. The sequence was inserted into the vector and transformed into Agrobacterium to verify the function preliminarily. The DNA sequence of 754 bp was amplified. The MdGA3ox1 gene promoter contained the basic elements: TATA-box, CAAT-box and other induced elements: light, gibberellin, enthylene and abscisic acid responsive elements. In order to verify the function of the sequence, the DNA fragments was inserted into the binary vector pCAMBIA1381 and named p1381-GA3ox1p. The plasmid of p1381-GA3ox1p was transformed into Agrobacterium EHA105. The analysis of GUS staining showed the promoter could drive the expression of GUS in transgenic Agrobacterium. The upstream sequence of MdGA3ox1 which contained the cis-elements with GA regulation was obtained. It also confirmed that the sequence had the promoter function.