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A novel bi-directional promoter cloned from melon and its activity in cucumber and tobacco

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Abstract

A bi-directional promoter, DP, was cloned by PCR amplification using the genomic DNA of melon as template. Analysis of itscis-acting elements in both directions revealed a series of inducible regulatory elements and some enhancer elements. To evaluate its transcriptional activity, DP in both directions was then cloned into vector pBI121 to replace the CaMV 35S promoter. DP in both directions also was inserted downstream of CaMV 35S to investigate whether the double promoter might affect expression of theuidA reporter gene at higher levels. Transient expression in cucumber leaves, stems, and fruits as well as in tobacco leaves and stems showed that DP in both directions drove transcription to much higher levels than did the single promoter CaMV 35S. However, activity of the double promoter was lower than the corresponding activity of the single promoter DP in both directions. These results demonstrate that DP is a natural bi-directional promoter, with much more activity than is found with the CaMV 35S promoter. Furthermore, in cucumber and tobacco, it is not suitable to insert DP in either direction downstream of the CaMV 35S promoter to form a double promoter.

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Authors and Affiliations

  1. Key Laboratory for Bioactive Material of Ministry of Education, Institute of Molecular Biology Nankai University, 300071, Tianjin, China

    Cuiyan Wang, Dongfeng Ding, Ruixiang Yan, Xiaoju Yu & Minggang Li

  2. National Engineering and Technology Research Center for Agricultural Products Freshness Protection, 300384, Tianjin, China

    Ruixiang Yan

  3. The University of Tokyo, Tokyo, Japan

    Weidong Li

Authors
  1. Cuiyan Wang
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  2. Dongfeng Ding
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  3. Ruixiang Yan
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  4. Xiaoju Yu
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  5. Weidong Li
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  6. Minggang Li
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Correspondence to Minggang Li.

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Wang, C., Ding, D., Yan, R. et al. A novel bi-directional promoter cloned from melon and its activity in cucumber and tobacco. J. Plant Biol. 51, 108–115 (2008). https://doi.org/10.1007/BF03030719

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  • DOI: https://doi.org/10.1007/BF03030719

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