Factors influencing efficiency of transient gene expression in the red macrophyte Porphyra yezoensis
Factors influencing efficiency of transient gene expression in the red macrophyte Porphyra yezoensis
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DOI：
10.1016/j.plantsci.2007.12.006
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发表时间：
2008-03-01
期刊：
影响因子：
5.2
通讯作者：
Saga, Naotsune
中科院分区：
文献类型：
--
作者：
Fukuda, Satoru;Mikami, Koji;Saga, Naotsune
海洋红藻紫菜（Porphyra yezoensis）由于其在生物学和经济上的重要性，已被提出作为海藻生理学和遗传学研究的模式植物。然而，利用基因转染和遗传转化系统进行分子生物学研究的进展一直受到外源基因在叶藻细胞中表达困难的阻碍。为了克服这种情况，我们开发了一种瞬时基因表达系统来监测叶藻细胞中的基因表达。合成了一个人工β -葡糖醛酸酶（GUS）编码区，使其适应yezoensis （PyGUS）密码子的使用，并通过粒子轰击评估了其作为瞬时基因表达报告者的效率。我们还证明了利用叶红油菜甘油醛-3-磷酸脱氢酶（GAPDH）基因启动子高效表达PyGUS基因的重要性，因为花椰菜花叶病毒（CaMV） 35S启动子在叶红油菜细胞中活性较低，而该启动子已成功用于监测高等植物细胞核和叶绿体中的基因表达。因此，缺乏对yezoensis和陆生植物之间基因表达调控机制差异的了解似乎是以前没有在yezoensis中开发出监测基因表达的实验系统的原因。在叶藻中建立瞬时基因表达系统有助于叶藻生物技术的发展。2007爱思唯尔爱尔兰有限公司版权所有。
The marine red alga Porphyra yezoensis has been proposed as a model plant for physiological and genetic studies in seaweeds because of its biological and economical importance. However, the progress of molecular biological studies using gene transfection and genetic transformation systems has been hindered by difficulties in the expression of foreign genes in P. yezoensis cells. To overcome this situation, we developed a transient gene expression system to monitor gene expression in P. yezoensis cells. An artificial beta-glucuronidase (GUS) coding region was synthesized to adapt it to the codon usage of P. yezoensis (PyGUS) and then evaluated for efficiency as a reporter of transient gene expression by particle bombardment. We also demonstrated the importance of using the promoter of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene from P. yezoensis for efficient expression of PyGUS, because the cauliflower mosaic virus (CaMV) 35S promoter, which has been successfully used for monitoring gene expression in nuclei and chloroplasts of higher plants, was less active in P. yezoensis cells. Therefore, the lack of knowledge about differences in the regulatory machinery of gene expression between P. yezoensis and terrestrial plants seems to be why experimental systems for monitoring gene expression were previously not developed in P. yezoensis. Establishment of the transient gene expression system in P yezoensis could facilitate biotechnological developments in this organism. (c) 2007 Elsevier Ireland Ltd. All rights reserved.