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Development of an expression system using the heat shock protein 70 promoter in the red macroalga, Porphyra tenera

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Abstract

Porphyra is a commercially valuable source of food and drugs and an important model organism for algal research. However, genetic research on Porphyra tenera has been limited by a lack of a heterologous gene expression system. In this study, we isolated native promoter PtHSP70 for the efficient expression of foreign genes in this organism. This promoter lies approximately 1 kb upstream of the heat shock protein 70 coding sequence and was isolated using adapter ligation-mediated genomic polymerase chain reaction. Promoter activity was evaluated using the synthetic GUS gene (PyGUS) with optimized codons for Porphyra yezoensis. Interestingly, the PtHSP70 promoter allowed the efficient expression of PyGUS in P. tenera and P. yezoensis, whereas the PyGAPDH promoter from P. yezoensis was not fully functional in P. tenera. The PtHSP70 promoter may have a more conserved regulatory mechanism than the PyGAPDH promoter between these species, suggesting that PtHSP70 could serve as a universal promoter for Porphyra species. We also established an efficient transient transformation system for P. tenera by evaluating transformation parameters including gold particle quantity, helium and vacuum pressure, developmental stages of leafy gametophytes, and target distance. Under optimal conditions of transient transformation, the frequency of GUS expression was determined by histochemical staining as 30–50 cells per bombardment. In addition, PyGUS expression was detected during the regeneration of monospores in P. tenera, indicating successful genetic transformation. Therefore, the new transient transformation system using the PtHSP70 promoter can be used for foreign gene expression in P. tenera, which may advance the development of P. tenera as a model organism.

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Acknowledgments

This research was supported by a grant (RP-2010-AQ) from National Fisheries Research and Development Institute, Korea, a grant from the Advanced Biomass R&D Center (ABC) of Korea Grant funded by the Ministry of Education, Science and Technology (ABC-2010-0029723), and a grant from the KRIBB Research Initiative Program.

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Authors and Affiliations

  1. Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 111 Gwahangno, Yuseong-gu, Daejeon, 305-806, South Korea

    Su Hyun Son, Joon-Woo Ahn, Jang Ryol Liu & Won-Joong Jeong

  2. Department of Biology, Kunsan National University, 1170 Daehangno, Gunsan, 573-701, South Korea

    Su Hyun Son & Dongsu Choi

  3. Graduate School of Fisheries Sciences, Hokkaido University, 3-1-1 Minato, Hakodate, 041-8611, Japan

    Toshiki Uji

  4. Department of Biology Education, Chonnam National University, Kwagnju, 500-757, South Korea

    Dong-Woog Choi

  5. Seaweed Research Center, National Fisheries Research and Development Institute, Mokpo, Jeonnam, 530-831, South Korea

    Eun-Jeong Park & Mi Sook Hwang

  6. Faculty of Fisheries Sciences, Hokkaido University, 3-1-1 Minato, Hakodate, 041-8611, Japan

    Koji Mikami

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  1. Su Hyun Son
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  2. Joon-Woo Ahn
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  3. Toshiki Uji
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  5. Eun-Jeong Park
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  7. Jang Ryol Liu
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  8. Dongsu Choi
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  9. Koji Mikami
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  10. Won-Joong Jeong
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Correspondence to Won-Joong Jeong.

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Su Hyun Son and Joon-Woo Ahn contributed equally to this work.

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Son, S.H., Ahn, JW., Uji, T. et al. Development of an expression system using the heat shock protein 70 promoter in the red macroalga, Porphyra tenera . J Appl Phycol 24, 79–87 (2012). https://doi.org/10.1007/s10811-011-9652-9

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  • DOI: https://doi.org/10.1007/s10811-011-9652-9

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