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.
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.