For tissue specific rescue experiments, mig-21
genomic sequence was amplified using 5-ATGGAGCGCGACTCTAAC-3 and 5-TCACCAAAAACTTTCATCTTTCGGG-3 and sub-cloned into pJET1.2. To obtain Pegl-17, 4.6 kb of sequence upstream of the egl-17
coding sequence was PCR amplified. Next, Pegl-17, mig-21
and the unc-54
-3 UTR were cloned using Gateway technology into pDONRP4-P1R, pDONR221 and pDONRP2R-P3 entry clones respectively. Pegl-17::mig-21
-3UTR (pKN162) was generated by 3-way Gateway cloning into the destination vector pKN133.