Fig. 2. Isolation of the ROXY1 gene. (A) T-DNA insertion sites are shown for each allele. The T-DNA insert in roxy1-1 is located in the promoter, 175 bp upstream of the start codon ATG. roxy1-2 and roxy1-3 carry insertions in the single coding exon, 91 and 291 bp downstream of the ATG, respectively. Numbering of coordinates is based on the Arabidopsis genomic clone BACF1C9 that contains the Col-0 ROXY1 gene. Left and right (L and R) T-DNA borders are only indicated where they could be identified. (B) ROXY1 cDNA and predicted amino acid sequence encoding a 136 amino acid protein (GenBank Accession Number AY910752). The dithiol CCMC and the monothiol CSGS motif are underlined. Locus name is At3g02000. (C) Comparison of ROXY1 expression in wild-type and roxy1-2 and roxy1-3 mutants by RT-PCR. RNA was isolated from inflorescences and primers were used allowing specific cDNA amplification. As a control, reactions with 18S rRNA were carried out.