HUSCAP logo Hokkaido Univ. logo

Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences >
Peer-reviewed Journal Articles, etc >

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

Files in This Item:
Son et al 20100805-2.pdf987.65 kBPDFView/Open
Please use this identifier to cite or link to this item:http://hdl.handle.net/2115/48276

Title: Development of an expression system using the heat shock protein 70 promoter in the red macroalga, Porphyra tenera
Authors: Son, Su Hyun Browse this author
Ahn, Joon-Woo Browse this author
Uji, Toshiki Browse this author →KAKEN DB
Choi, Dong-Woog Browse this author
Park, Eun-Jeong Browse this author
Hwang, Mi Sook Browse this author
Liu, Jang Ryol Browse this author
Choi, Dongsu Browse this author
Mikami, Koji Browse this author →KAKEN DB
Jeong, Won-Joong Browse this author
Keywords: Transient gene expression
Particle bombardment
Heat shock protein 70 (HSP70)
Porphyra tenera
Issue Date: Jan-2012
Publisher: Springer
Journal Title: Journal of Applied Phycology
Volume: 24
Issue: 1
Start Page: 79
End Page: 87
Publisher DOI: 10.1007/s10811-011-9652-9
Abstract: Porphyra is a commercially valuable source of food and drugs, and represents an important model organism for algal research. However, genetic research on P. tenera has been limited due to lack of a heterologous gene expression system. In the present study, we isolated a native promoter, the PtHSP70 promoter, for efficient expression of foreign genes in this organism. This promoter lies approximately 1 kb upstream of the coding sequence for Heat Shock Protein 70 (HSP70) and was isolated using adapter-mediated genomic PCR. Promoter activity was evaluated using the synthetic GUS gene (PyGUS) with optimized codons for Porphyra yezoensis. Interestingly, the PtHSP70 promoter allowed equivalent expression of PyGUS in both P. tenera and P. yezoensis, whereas the GAPDH promoter from P. yezoensis was not fully functional in P. tenera. These data suggest that the PtHSP70 promoter has a more conserved regulatory mechanism than the PyGAPDH promoter between these species. We also established an efficient transient transformation system for P. tenera by evaluating various transformation parameters such as quantity of gold particles, pressure of helium and vacuum, developmental stages of leafy gametophytes, and target distance. Under the optimal conditions of transient transformation, the frequency of GUS expression was determined by histochemical staining to be 30-50 cells per bombardment. 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.
Rights: The original publication is available at www.springerlink.com
Type: article (author version)
URI: http://hdl.handle.net/2115/48276
Appears in Collections:水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 三上 浩司

Export metadata:

OAI-PMH ( junii2 , jpcoar )

MathJax is now OFF:


 

 - Hokkaido University