-
Methods
Strain production
Lines were generated by transposition of P{GawB}
element of Brand
and Perrimon (1993), mapped and balanced according to the protocol
by Yoshihara and Ito (2000). Some of
the lines were balanced directly with strains containing second and
third chromosome balancers.
Gal4 expression
analyses
Embryo: Males
of each strain were crossed to w; UAS-gfpnlacZ UAS-gfp[S65T] females
and embryos were collected and fixed according to the method by Hummel
et al. (Dev. Genes & Evol. 207: 131-135). Embryos
were stained with anti-beta galactosidase monoclonal antibody and ABC
elite kit and mounted in aqueous mountant (Mowiol) for microscopic observation.
Larva:
Dissected larvae were fixed and stained for beta galactosidase activity
with X-gal. Stained imaginal discs were mounted for microscopic observation.
Live larvae were also observed directly under dissecting GFP microscope
(Leika).
Adult:
Some of adult flies were observed under dissecting GFP microscope.
Inverse PCR and
sequencing
DNA purification
Inverse PCR
DNA sequencing
Blast Search,
Mapping and Database Construction
Blast search was performed by the program package
Dynaclust lite TM against Release 1.0 Cerela/BDGP sequence. Database
was constructed by Sybase platform.
- Contributor
Strain
production, maintenance and expression analyses in NIG
Misako Taniguchi, Hiroko Takeuchi, Yuko Kurita,
Hiromi Niwa, Reiko Iwoki,
Qun Rei, Sachiko Yoneyama, Kazumi Tanimoto, Aya Matsunaga, Miyuki Kobari,
Yuki Kagoshima.
Inverse
PCR Mapping
Yukiko Sado, Misako Taniguchi, Ai Akimoto
DNA sequencing
Yuji Kohara (NIG, Genetic Resource Center)
- Acknowledgement
This project in Hayashi lab was supported by Japan Society for Promotion
of Science (Research for Future), and by Grant-in-Aid for Scientific
Research on Priority Areas (C) "Genome Science" from the Ministry
of Education, Culture, Sports, Science and Technology of Japan.
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