Transient Gene Expression System Established in Porphyra yezoensis Is Widely Applicable in Bangiophycean Algae

Abstract

The establishment of transient gene expression systems in the marine red macroalga Porphyra yezoensis has been useful for the molecular analysis of cellular processes in this species.However, there has been no successful report about the expression of foreign genes in other red macroalgae, which has impeded a broader understanding of the molecular biology of these species. We therefore examined whether the P. yezoensis transient gene expression system was applicable to other red macroalgae. The results indicated that a codon-optimized GUS,

Transient Transformation of Red Algal Cells: Breakthrough Toward Genetic Transformation of Marine Crop Porphyra Species

“…1 and 2), indicating the enhancement of the expression level of the GUS reporter gene. Optimization of the codon usage of the reporter gene is therefore one of the important factors for successful expression in P. yezoensis cells (Fukuda et al, 2008;Mikami et al, 2009;Takahashi et al, 2010;Uji et al, 2010;Hirata et al, 2011).…”

“…Thus, it was concluded that the transient gene expression system developed in P. yezoensis is widely applicable in Bangiophycean red algae . The number of reporter-expressing cells varies among species (Table 2; Hirata et al, 2011), suggesting the possibility of the improvement of the gene transfer and expression systems in each species using respective endogenous strong promoters to drive the expression of the PyGUS and the sGFP(S65T) reporter genes. Similarly, expression of these reporter genes in Florideophycean algae seems to be possible if suitable endogenous promoters can be employed.…”

Advances in genetic engineering of marine algae

“…The duplicated carbonic anhydrase 1 (DCY1) promoter was identified and used for stable nuclear transformation in D. salina (Li et al, 2010, Lu et al, 2011. The endogenesis PyAct1 (5′ upstream region of the actin1 gene from P. yezoensis) promoter was also confirmed to be effective in the transient gene expression of 12 red seaweed species (Hirata et al, 2011, Takahashi et al, 2010. In Nannochloropsis sp.…”

“…The GUS gene encoding the β-glucuronidase is typically selected as an effective reporter for transient and stable expression in marine algae, e.g., D. salina (Tan et al, 2005), Amphidinium sp., Symbiodinium microadriaticum (ten Lohuis and Miller, 1998), T. weissflogii (Falciatore et al, 1999), Ectocarpus sp. (Cheney and Kurtzman, 1992), Porphyra yezoesis (Hirata et al, 2011, Kuang et al, 1998, Liu et al, 2003, Ulva lactuca (Huang et al, 1996), L. japonica , Qin et al, 1994, Undaria pinnatifida (Qin et al, 1994). However, because the substrate of this gene has an impermeable membrane, it is toxic to the transformed cell and damages the cellular ultrastructure during the dyeing process.…”

Identification and Efficient Utilization of Antibiotics for the Development of a Stable Transformation System in Porphyra yezoensis (Bangiales, Rhodophyta)

“…To date, several studies have described the establishment of a transient gene expression system in P. yezoensis [9]; for example, we have succeeded in eliciting the efficient expression of the codon-modified β-glucuronidase (PyGUS) and humanized cyan and green fluorescent protein (AmCFP and ZsGFP) genes by transient transformation of P. yezoensis gametophytes using particle bombardment [10][11][12][13], which was then adapted to other Bangiophycean algae [14,15]. Despite the development of transient gene expression systems in P. yezoensis, a stable transformation system, which is needed for the fine analysis of gene function, has not been established, because of a lack of techniques to select and isolate stably transformed cells from gametophytes.…”