Tagged Mutagenesis and Gene-trap in the Moss, Physcomitrella patens by Shuttle Mutagenesis |
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Authors: | Nishiyama Tomoaki; Hiwatashi Yuji; Sakakibara Keiko; Kato Masahiro; Hasebe Mitsuyasu |
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Institution: | 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 |
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Abstract: | The moss, Physcomitrella patens has been used as a useful materialin many fields, because of its simple body plan, ease of genetargeting, and other reasons. Although many mutants have beenreported, no method to isolate the corresponding genes was reported.We developed a gene tagging and gene-trap system in P. patensby using the shuttle mutagenesis technique, which has been usedin the budding yeast. In 5264 tagged lines, 203 mutants withaltered developmental or morphological phenotypes were obtained.In 129 of 4757 gene-trap lines, ß-glucuronidase (GUS)activity was detected in some tissue. Although multiple copiesof a tag were detected in many tagged lines by Southern analyses,most copies are likely integrated at the same locus accordingto PCR analyses. |
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Keywords: | tagging gene-trap Physcomitrella patens homologous recombination shuttle mutagenesis |
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