Heterologous activation of the Porphyra tenera HSP70 promoter in Bangiophycean algal cells.

Ryo Hirata, Won-Joong Jeong, Naotsune Saga, Koji Mikami
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引用次数: 8

Abstract

Porphyra has attracted great attention for its biological and industrial importance. However, establishment of a stable nuclear transformation has not yet been achieved in these organisms, which impedes the molecular biological study and the development of a molecular breeding method for them. Toward establishing the stable transformation, we have recently developed an efficient transient gene expression system in Bangiophycean algae, in which the HSP70 promoter from P. tenera (PtHSP70 promoter) was activated heterologously in P. yezoensis cells. Since heterologous promoters are required for homologous recombination-based stable transformation, the identification of heterologously activated promoters is important in establishing a stable transformation system in individual Bangiophycean alga. We here examined the activation of the PtHSP70 promoter using the GC-rich PyGUS reporter system in additional Porphyra and Bangia species. The results indicated that this promoter drove expression of the PyGUS gene efficiently in all examined algae, whereas there was quite low expression of PyGUS by the cauliflower mosaic virus 35S promoter that is widely used as a heterologous promoter in the transformation of green land plants. Therefore, heterologous activation of the PtHSP70 promoter could promote the establishment of the stable transformation system in various kinds of Bangiophycean algae.
斑藻细胞中卟啉HSP70启动子的异源激活。
紫菜因其在生物和工业上的重要性而受到广泛关注。然而,在这些生物中尚未建立稳定的核转化,这阻碍了分子生物学研究和分子育种方法的发展。为了建立稳定的转化,我们最近开发了一个高效的瞬时基因表达系统,该系统在P. yezoensis细胞中异源激活来自P. tenera的HSP70启动子(PtHSP70启动子)。由于同源重组稳定转化需要异源启动子,因此鉴定异源激活启动子对于在bangiophyean藻类个体中建立稳定转化体系具有重要意义。我们利用富含gc的PyGUS报告系统,在另外的卟啉和Bangia物种中检测了PtHSP70启动子的激活。结果表明,该启动子在所有被试藻类中都能有效地驱动PyGUS基因的表达,而在绿地植物转化中广泛用作异源启动子的花椰菜花叶病毒35S启动子对PyGUS基因的表达水平很低。因此,异源激活PtHSP70启动子可以促进各种bangiophyan藻类中稳定转化体系的建立。
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