Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, 041-8611, Japan.
Mar Biotechnol (NY). 2011 Oct;13(5):1038-47. doi: 10.1007/s10126-011-9367-6. Epub 2011 Feb 1.
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 the 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, designated PyGUS, and plant-adapted sGFP(S65T) were successfully expressed under the control of the P. yezoensis PyAct1 promoter in gametophytic cells of six Porphyra species and also in Bangia fuscopurpurea, all of which are classified as Bangiophyceae. In contrast, there were no reporter-expressing cells in the Florideophycean algae examined. These results indicate the availability of PyGUS and sGFP as reporters and the 5' upstream region of the PyAct1 gene as a heterologous promoter for transient gene expression in Bangiophycean algae, which could provide a clue to the efficient expression of foreign genes and transformation in marine red macroalgae.
在海洋红藻紫菜中建立瞬时基因表达系统对于该物种细胞过程的分子分析非常有用。然而，关于外源基因在其他红藻中的表达还没有成功的报道，这阻碍了对这些物种分子生物学的更广泛理解。因此，我们研究了紫菜瞬时基因表达系统是否适用于其他红藻。结果表明，一种密码子优化的 GUS，命名为 PyGUS，以及植物适应的 sGFP(S65T)，在 PyAct1 启动子的控制下，在六个紫菜物种的配子体细胞中以及在 Bangia fuscopurpurea 中成功表达，所有这些都被归类为红藻门。相比之下，在检查的 Florideophycean 藻类中没有报告基因表达细胞。这些结果表明 PyGUS 和 sGFP 可作为报告基因，PyAct1 基因的 5'上游区可作为异源启动子，用于 Bangiophycean 藻类中的瞬时基因表达，这可能为海洋红藻中外源基因的高效表达和转化提供线索。
