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

Cited 16 time in scopus
Metadata Downloads
Title
Development of an expression system using the heat shock protein 70 promoter in the red macroalga, Porphyra tenera
Author(s)
S H Son; Joon Woo Ahn; T Uji; D W Choi; E J Park; M S Hwang; Jang Ryol Liu; D Choi; K Mikami; Won Joong Jeong
Bibliographic Citation
Journal of Applied Phycology, vol. 24, no. 1, pp. 79-87
Publication Year
2012
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.
Keyword
Heat shock protein 70HSP70Particle bombardmentPorphyra teneraTransient gene expression
ISSN
0921-8971
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s10811-011-9652-9
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Cell Factory Research Center > 1. Journal Articles
Files in This Item:
  • There are no files associated with this item.


Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.