Isolation and functional characterization of bidirectional promoters in rice
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 microar...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2016-05-01
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| Series: | Frontiers in Plant Science |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fpls.2016.00766/full |
| _version_ | 1818878121741385728 |
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| author | Rui eWang Yan eYan Menglin eZhu Mei eYang Fei eZhou Hao eChen Yongjun eLin |
| author_facet | Rui eWang Yan eYan Menglin eZhu Mei eYang Fei eZhou Hao eChen Yongjun eLin |
| author_sort | Rui eWang |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-19T14:09:09Z |
| format | Article |
| id | doaj.art-2c270c64fe954567aa8c8a322967f154 |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-12-19T14:09:09Z |
| publishDate | 2016-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-2c270c64fe954567aa8c8a322967f1542022-12-21T20:18:12ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2016-05-01710.3389/fpls.2016.00766197106Isolation and functional characterization of bidirectional promoters in riceRui eWang0Yan eYan1Menglin eZhu2Mei eYang3Fei eZhou4Hao eChen5Yongjun eLin6Huazhong Agricultural UniversityChinese Academy of Tropical Agricultural SciencesHuazhong Agricultural UniversityHuazhong Agricultural UniversityHuazhong Agricultural UniversityHuazhong Agricultural UniversityHuazhong Agricultural UniversityBidirectional 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.http://journal.frontiersin.org/Journal/10.3389/fpls.2016.00766/fullricestable transformationbidirectional promoterconservation analysisGUS assayGFP assay |
| spellingShingle | Rui eWang Yan eYan Menglin eZhu Mei eYang Fei eZhou Hao eChen Yongjun eLin Isolation and functional characterization of bidirectional promoters in rice Frontiers in Plant Science rice stable transformation bidirectional promoter conservation analysis GUS assay GFP assay |
| title | Isolation and functional characterization of bidirectional promoters in rice |
| title_full | Isolation and functional characterization of bidirectional promoters in rice |
| title_fullStr | Isolation and functional characterization of bidirectional promoters in rice |
| title_full_unstemmed | Isolation and functional characterization of bidirectional promoters in rice |
| title_short | Isolation and functional characterization of bidirectional promoters in rice |
| title_sort | isolation and functional characterization of bidirectional promoters in rice |
| topic | rice stable transformation bidirectional promoter conservation analysis GUS assay GFP assay |
| url | http://journal.frontiersin.org/Journal/10.3389/fpls.2016.00766/full |
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