A 9080 bp XbaI-SmaI fragment of T26E3, representing par-6
, was subcloned into the pBluescript KS+ vector (Stratagene). This subclone was predicted to include the entire coding region, 3'-untranslated region and 3-5 kb of sequence 5' of the gene. A PstI site was introduced either just before or in the stop codon of the gene by site-directed mutagenesis (Quickchange kit, Stratagene) or recombinant PCR. To construct par-6
::gfp the coding region of gfp was amplified by PCR from plasmid pPD95.75 (1995 Fire Lab vector kit, www.ciwemb.edu) and cloned into the introduced PstI site.