Operation of an efficient site-specific recombination system of Zygosaccharomyces rouxii in tobacco cells

Hitoshi Onouchi, Kumi Yokoi, Chiyoko Machida, Hiroaki Matsuzaki, Yasuji Oshima, Ken Matsuoka, Kenzo Nakamura, Yasunori Machida

研究成果: ジャーナルへの寄稿学術誌査読

98 被引用数 (Scopus)

抄録

Recombinase encoded by the R gene of pSR1 of Zygosaccharomyces rouxii mediates reciprocal recombination between two specific recombination sites (RSs) to induce excision or inversion of the DNA segment that is flanked by the RSs. We report here that site-specific recombination mediated by this system takes place efficiently in tobacco cells. To monitor the recombination events in tobacco cells, we have constructed two types of cryptic β-glucuronidase reporter gene in such a way that recombination such as inversion of the construct or excision of the intervening sequence results in their expression. When these cryptic reporter constructs were transiently introduced together with the R gene by electroporation into protoplasts of tobacco cells, β-glucuronidase activity was detected. The cryptic reporter genes, when stably resident in the chromosome of tobacco cells, were also activated by the R gene. Structural analyses of the genomic DNA isolated from these tobacco cells showed that the R protein did in fact catalyze precise recombination between two copies of RSs in tobacco cells, with resultant activation of the cryptic reporter genes. This observation provides the basis for development of a DNA technology whereby large regions of DNA can be manipulated in plant chromosomes. Potential uses of this recombination system are discussed.

本文言語英語
ページ(範囲)6373-6378
ページ数6
ジャーナルNucleic acids research
19
23
DOI
出版ステータス出版済み - 12月 11 1991
外部発表はい

!!!All Science Journal Classification (ASJC) codes

  • 遺伝学

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