Figure 2.

Genetic organization and expression of C. elegans catp-5. A) C. elegans N2 wild-type and norspermidine-resistant nor-2 mutant worms were analyzed for their genotype by single-worm PCR analyses. A part of the gene catp-5 was amplified by using oligonucleotides flanking the deletion present in the catp-5(mun1) allele of nor-2 (see Materials and Methods). B) RT-PCR analyses of C. elegans N2 wild-type and catp-5(mun1) animals. Oligonucleotides flanking the deletion of catp-5 were used in the PCR reaction (see Materials and Methods). No catp-5 PCR product was obtained for the catp-5(mun1) allele. Owing to the deletion identified at the genomic level, the theoretical size of the catp-5 PCR fragment in catp-5(mun1) worms is 612 bp. Band of ∼900 bp corresponds to the internal control PCR fragment of the act-1 gene. C) Genomic organization of C. elegans catp-5 and reporter gene constructs used in microinjection. Position of the deletion allele catp-5(mun1) identified in the nor-2 strain is shown. D) Expression pattern of C. elegans CATP-5. Transgenic animals carrying the construct catp-5(3.9)::gfp (a) or catp-5(6.2)::gfp (b) exhibited cytosolic GFP signals with the same spatial and temporal expression pattern. In transgenic animals carrying catp-5(12.2)::gfp (c–h), which includes the complete ORF of catp-5, fluorescent signals are associated with the excretory cell (e, f) and the apical membrane of the intestinal cells (g, h). Scale bars = 100 μm (a–d); 50 μm (e–h).