桃PpMADS1基因启动子的克隆及功能分析

中国生物工程杂志

2010, Vol. 30

Issue (05): 57-62

研究报告

桃PpMADS1基因启动子的克隆及功能分析

李翠¹,谢华^1,2,徐勇²,李云伏^1,2,马荣才^1,2*

1.首都师范大学生命科学学院北京 1000482
2.北京农林科学院农业生物技术研究中心北京 100097

Cloning and Function Analysis of the Promoter of PpMADS1 in Peach

LI CuiXIE Hua^1,2, XU Yong²,LI Yun-fu^1,2,MA Rong-cai^1,2

1.College of Life Science, Capital Normal University, Beijing 100048,China
2.Beijing AgroBiotechnology Research Centre，Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,China

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摘要：

PpMADS1基因属于一类MADS box 基因，在植物的花发育调控中起着重要的作用。通过Genome Walking的方法从桃基因组中分离了长度为1 814bp的PpMADS1基因启动子片段，序列分析表明，在此启动子上不仅含有TATA box 和CAAT box基本元件，而且含有大量的与光调节有关的调控元件，如GT-1，Sp1和as-2-box，另外存在两个CArG-box元件、一个G-box元件和一个TGA-element，说明该启动子可能受光周期和激素的调控。将该启动子通过5′端缺失，分区段与GUS报告基因连接构建表达载体，并转化拟南芥。GUS组织化学染色分析结果表明，在-197到-454bp有促使GUS在花原基中表达的花原基特异性元件，在-454到-678bp之间存在促使GUS在萼片和花瓣表达的特异性元件，在-678到-978bp存在负调控作用元件，阻遏了GUS基因在花药中的表达。

关键词： 桃; 花发育; PpMADS1; 启动子; 顺式作用元件

Abstract:

In order to analyze transcriptional regulative mechanism of PpMADS1 gene, the PpMADS1 promoter was obtained using Genome Walking method from peach (Prunus persica) genomic DNA. Sequence analysis indicated that there exist TATA-box, CAAT-box, two CArG box, one G-box, one TGA-element and a large number of regulatory elements involved in light response, such as GT-1, Sp1 and as-2-box, these results indicated that the promoter may be regulated by light and hormone. The PpMADS1 promoter was truncated according to the prediction of putative of cis-acting elements and fused with GUS reporter gene and transferred into Arabidopsis thaliana. Histochemical staining of different organs of the transgenic plants showed that the region between -197 to -454bp specified GUS expression in flower primordium and the region between -454 to -678bp in sepals and petals. A negatively regulatory element was shown to be present between -678 to -978bp that repressed GUS expression in filament.

Key words: development PpMADS1 Promoter cis element Floral

收稿日期: 2010-01-20 出版日期: 2010-05-25

通讯作者: 马荣才 E-mail: marc@bjny.gov.cn

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引用本文:

李翠谢华徐勇李云伏马荣才. 桃PpMADS1基因启动子的克隆及功能分析[J]. 中国生物工程杂志, 2010, 30(05): 57-62.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I05/57

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