Preface
for PHYSCOmanual
The moss Physcomitrella
patens is
an emerging model organism for various fields in science, and it will be more
extensively used in near future. There should be several advantages to use this
moss, in which the most prominent is its feasibility in gene targeting based on
the high homologous recombination rate similar to that of fission yeast. P.
patens
nuclear genome will be fully sequenced and annotated in a couple of years,
which will facilitate the reverse genetic approaches. Forward genetics are also
applicable for this moss and other new techniques especially on imaging at
cellular level are enormously improved recently. We intend this laboratory
manual to be useful for new in P. patens research. Furthermore,
we hope that this laboratory manual will be useful also for more established
researches to improve their experimental techniques. Actually, contents of this
manual are extensively changed in the last few years.
This
manual starts from the cultivation and the observation of wild type moss. Then,
necessary techniques for reverse genetics, such as gene isolation, southern
hybridization, northern hybridization, and gene targeting are described in
detail. These are mostly similar to regular techniques in molecular biology,
but there are some minor but critical tips for P. patens, which are
stressed in the text. Useful techniques to characterize gene functions, such as
the observation of disruptant and overexpressor phenotypes and the detection of
protein localization, were fully described. The last chapter focuses how to use
PHYSCObase. Many plasmids listed at the end of this manual are all available.
Japanese
version of this manual was originally made to share laboratory techniques in
our laboratory, and then opened for public in our web site since 1998. All
protocols were written by former and present members in our laboratory, and I
deeply appreciate for their enthusiasm to Physcomitrella patens. Rumiko
Kofuji (Faculty of Science, Kanazawa University), Keiko Sakakibara (Graduate School
of Science, Hiroshima University), Tomoaki Nishiyama (Advanced Science Research
Center, Kanazawa University), and Tomomichi Fujita (Graduate School of
Sciences, Hokkaido University) left my laboratory, and are now expanding moss
biology intheir new laboratories. All authors of this manual appreciate the
open-minded Physco community for sharing knowledge and many techniques,
especially David Cove, Didier Schaefer, Jean-Pierre Zryd, Andy Cumming, Celia
Knight, Ralf Reski, Ralph Quatrano and more moss friends. The old version was
expanded to cover a broader range of techniques for the laboratory course in
National Institute for Basic Biology, and revised year by year.
We hope this manual will be useful for moss
researchers, and any comments and suggestions on the improvement of this manual
are very welcome.
Mitsuyasu
Hasebe