© 2000 by Kazusa DNA Research Institute
Tagged Mutagenesis and Gene-trap in the Moss, Physcomitrella patens by Shuttle Mutagenesis
1Department of Biological Sciences, Graduate School of Science, the University of Tokyo Tokyo 113-0033, Japan
2National Institute for Basic Biology Okazaki 444-8585, Japan
3Department of Molecular Biomechanics, The Graduate University for Advanced Studies Okazaki 444-8585, Japan
4PRESTO, Japan Science and Technology Corporation
* To whom correspondence should be addressed. Tel./Fax. +81-564-55-7546, E-mail: mhasebe{at}nibb.ac.jp
The moss, Physcomitrella patens has been used as a useful material in many fields, because of its simple body plan, ease of gene targeting, and other reasons. Although many mutants have been reported, no method to isolate the corresponding genes was reported. We developed a gene tagging and gene-trap system in P. patens by using the shuttle mutagenesis technique, which has been used in the budding yeast. In 5264 tagged lines, 203 mutants with altered developmental or morphological phenotypes were obtained. In 129 of 4757 gene-trap lines, ß-glucuronidase (GUS) activity was detected in some tissue. Although multiple copies of a tag were detected in many tagged lines by Southern analyses, most copies are likely integrated at the same locus according to PCR analyses.
Key words: tagging; gene-trap; Physcomitrella patens; homologous recombination; shuttle mutagenesis
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