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红球藻Cyanidioschyzon merolae 10D培养条件的改善及同源重组介导的核转化证据

Improvement of culture conditions and evidence for nuclear transformation by homologous recombination in a red alga, Cyanidioschyzon merolae 10D.

作者信息

Minoda Ayumi, Sakagami Rei, Yagisawa Fumi, Kuroiwa Tsuneyoshi, Tanaka Kan

机构信息

Institute of Molecular and Cellular Biosciences, The University of Tokyo, 1-1-1 Yayoi, 113-0032 Japan.

出版信息

Plant Cell Physiol. 2004 Jun;45(6):667-71. doi: 10.1093/pcp/pch087.

Abstract

Although the nuclear genome sequence of Cyanidioschyzon merolae 10D, a unicellular red alga, was recently determined, DNA transformation technology that is important as a model plant system has never been available thus far. In this study, improved culture conditions resulted in a faster growth rate of C. merolae in liquid medium (doubling time = 9.2 h), and colony formation on gellan gum plates. Using these conditions, spontaneous mutants (5-fluoroortic acid resistant) deficient in the UMP synthase gene were isolated. The lesions were then restored by introducing the wild-type UMP synthase gene into the cells suggesting DNA transformation by homologous recombination.

摘要

尽管单细胞红藻嗜热栖热放线菌10D的核基因组序列最近已被测定,但作为一种模式植物系统很重要的DNA转化技术迄今仍未获得。在本研究中,改良的培养条件使嗜热栖热放线菌在液体培养基中的生长速度加快(倍增时间 = 9.2小时),并在结冷胶平板上形成菌落。利用这些条件,分离出了UMP合酶基因缺陷的自发突变体(对5-氟乳清酸有抗性)。然后通过将野生型UMP合酶基因导入细胞来修复损伤,这表明通过同源重组进行了DNA转化。

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