Worm Breeder's Gazette 14(1): 22 (October 1, 1995)

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

Integration of extrachromosomal DNA arrays into a chromosome by UV- irradiation

Shohei Mitani

Dept. of Physiology, Tokyo Women's Medical College, 8-1, Kawada-cho, Shinjuku-ku, Tokyo, 162, Japan. FAX: 81-3-5269-7362.

To work with transgenic C. elegans animals, it is common to construct
strains with extrachromosomal DNA arrays by microinjection of DNA (Mello
et al., 1991). These strains, however, are mosaic in terms of the
transgene and sometimes need a second step to integrate the transgene into
a chromosome. The most common method to do this is to irradiate the parent
animals with gamma ray from a l37Cs source.

Because UV light induces the nematodes similar gene rearrangements with
those by gamma ray irradiation (Stewart et al., 1991), I developed a UV
irradiation method which may substitute a common gamma ray method for
integration of DNA into a chromosome of C. elegans.

Parent strains were irradiated with a UV light (254 nm) using a UV cross
linker (commonly used to fix polynucleotides to nylon membranes;
spectrolinker, Spectronics) at a dose of 300 J/m2 (the optimal dose among
examined [ranging from 150-1000 J/m2]). Irradiated parents were allowed to
lay eggs and healthy Fl animals were picked onto individual plates. F2
rollers from each plate were singled and strains which produced only
roller animals were identified later, as the gamma ray method. I have
found the probability of integration is high enough for routine
experiments: about 5%.

Since UV light is much easier to access for most researchers than the
gamma ray source, it appears a convenient alternative to obtain stable
transgenic animals with reporter gene fusion or heat-shock expression.