Operation of an efficient site-specific recombination system of Zygosaccharomyces rouxii in tobacco cells (original) (raw)
Journal Article
Search for other works by this author on:
Search for other works by this author on:
Search for other works by this author on:
2
Department of Fermentation Technology, Faculty of Engineering, Osaka University
2-1 Yamadaoka, suita-shi, Osaka 565, Japan
Search for other works by this author on:
2
Department of Fermentation Technology, Faculty of Engineering, Osaka University
2-1 Yamadaoka, suita-shi, Osaka 565, Japan
Search for other works by this author on:
1
Faculty of Agriculture, Nagoya University
Chikusa-ku, Nagoya 464-01
Search for other works by this author on:
1
Faculty of Agriculture, Nagoya University
Chikusa-ku, Nagoya 464-01
Search for other works by this author on:
* To whom correspondence should be addressed
Search for other works by this author on:
Received:
01 October 1991
Accepted:
13 November 1991
Published:
11 December 1991
PDF Cite
Hitoshi Onouchi, Kumi Yokoi, Chiyoko Machida, Hiroaki Matsuzaki, Yasuji Oshima, Ken Matsuoka, Kenzo Nakamura, Yasunori Machida, Operation of an efficient site-specific recombination system of Zygosaccharomyces rouxii in tobacco cells , Nucleic Acids Research, Volume 19, Issue 23, 11 December 1991, Pages 6373–6378, https://doi.org/10.1093/nar/19.23.6373
Close
Navbar Search Filter Mobile Enter search term Search
Abstract
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.
This content is only available as a PDF.
© 1991 Oxford University Press
I agree to the terms and conditions. You must accept the terms and conditions.
Submit a comment
Name
Affiliations
Comment title
Comment
You have entered an invalid code
Thank you for submitting a comment on this article. Your comment will be reviewed and published at the journal's discretion. Please check for further notifications by email.