Kage-Nakadai, Eriko et al. (2011) International Worm Meeting "Low-copy integration of transgenes by TMP/UV methods."

Yoshina, Sawako, Gengyo-Ando, Keiko, Kage-Nakadai, Eriko, Kobuna, Hiroyuki, Funatsu, Osamu, Mitani, Shohei, Ootori, Muneyoshi, Hori, Sayaka

[International Worm Meeting, 2011]

In Caenorhabditis elegans, transgenic strains are typically generated by injecting DNA into the germline to form high-copy extrachromosomal arrays. These transgenes are semi-stable and their expression are silenced in germline. In order to create single- or low-copy chromosomal integrated lines, methods using Mos1 transposon or microparticle bombardment have been developed. We have developed an alternative method by using TMP/UV, that produces low-copy integrations. We have successfully integrated low-copy of transgenes using positive selection with vps-45 rescue fragment / temperature sensitivity and negative selection with ben-1 rescue fragment / benomyl sensitivity. We confirmed that low-copy integrants express transgenes in germline. Currently, using this method, we are constructing a PhiC31-attBP / Cre-LoxP system.