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Isolation and Functional Characterization of Bidirectional Promoters in Rice

文献类型： 外文期刊

作者： Wang, Rui 1 ; Yan, Yan 3 ; Zhu, Menglin 1 ; Yang, Mei 1 ; Zhou, Fei 1 ; Chen, Hao 1 ; Lin, Yongjun 1 ;

作者机构： 1.Huazhong Agr Univ, Natl Key Lab Crop Genet Improvement, Wuhan, Peoples R China

2.Huazhong Agr Univ, Natl Ctr Plant Gene Res, Wuhan, Peoples R China

3.Chinese Acad Trop Agr Sci, Haikou, Hainan, Peoples R China

关键词： rice;bidirectional promoter;stable transformation;GUS assay;GFP assay;deletion analysis;conservation analysis

期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：5.753； 五年影响因子：6.612 ）

ISSN： 1664-462X

年卷期： 2016 年 7 卷

页码：

收录情况： SCI

摘要： Bidirectional promoters, which show great application potential in genetic improvement of plants, have aroused great research interest recently. However, most bidirectional promoters were cloned individually in the studies of single genes. Here, we initiatively combined RNA-seq data and cDNA microarray data to discover the potential bidirectional promoters in rice genome. Based on the expression level and correlation of each adjacent and oppositely transcribed gene pair, we selected four candidate gene pairs. Then, the intergenic region between each pair was isolated and cloned into a dual reporter vector pDX2181 for functional identification. GUS and GFP assays of the transgenic plants indicated that all the intergenic regions showed bidirectional expression activity in various tissues. Through 5' and 3' deletion analysis on one of the above bidirectional promoters, we identified the enhancing region which sharply increased its bidirectional expression efficiency and the essential regions respectively responsible for its 5' and 3' basic expression activity. The bidirectional arrangement of the four gene pairs in six gramineous plants was also analyzed, showing the conserved characteristics of the four bidirectional promoters identified in our study. In addition, two novel cis-sequences conserved in the four bidirectional promoters were discovered by bioinformatic identification. Our study proposes a feasible method for selecting, cloning, and functionally identifying bidirectional promoters as well as for discovering their bidirectional regulatory regions and conserved sequences in rice.

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