|
12th International Symposium on Bioluminescence & Chemiluminescence |
Symposium abstracts:
Yamauchi, Masatake, Sasanuma, Motoe, Tsuji, Satsuki, Terada, Minako, Ishikawa, Yuji
Radiation Safety Research Centre, National Institute of Radiological Sciences,
Chiba 263-8555 Japan.
Email: m_yam@nirs.go.jp
Medakafish (Oryzias latipes) belongs to a group of small oviparous teleost
fishes that inhabit fresh water in Asian countries including Japan. Experimental
procedures to introduce foreign genes into medakafish have been reported using
electroporation and/or microinjection. Microinjection is widely used in gene
transfer experiments. However, it requires high technical sophistication. Electroporation
can be easily applied to gene transfer experiments, but with a low efficiency.
Therefore, we attempted to develop an experimental procedure to introduce a
foreign gene without high technical sophistication but at reasonably high efficiency.
A particle gun is an instrument used to introduce DNA that is coated on gold
microcarrier particles directly into cells at high speed using high-pressure
gas. Here we report a procedure to introduce a foreign gene into fertilized
eggs of medakafish using the particle gun method. A plasmid construct with the
green fluorescence protein (GFP) gene driven by the medakafish beta-actin gene
promoter was successfully introduced into eggs, and the expression of GFP was
observed in 20% of the primary transfectant fish. In addition, germ line transmission
of GFP was observed in 13% of the GFP-positive primary transfectant.
This
is a preprint of an article accepted for publication in Luminescence: Copyright
2001 John
Wiley & Sons, Ltd (Wiley website)