Additional file 3: of Elevated growth temperature decreases levels of the PEX5 peroxisome-targeting signal receptor and ameliorates defects of Arabidopsis mutants with an impaired PEX4 ubiquitin-conjugating enzyme

2015-09-16T05:00:00Z (GMT) by Yun-Ting Kao Bonnie Bartel
PCR-based markers for genotyping mutants. pex4-1 carries a C-to-T mutation at the 533rd nucleotide that destroys an MnlΙ restriction enzyme cut site. pex4-2 carries an intronic G-to-A mutation at the 1085th nucleotide (18 bp upstream of the fourth exon of PEX4). We used the dCAPS website ( http://helix.wustl.edu/dcaps/dcaps.html , [73]) to design a genotyping marker for pex4-2. pex5-1 carries a C-to-T mutation at the 2910th nucleotide that destroys an EcoRΙ restriction enzyme cut site [3]. pex5-10 has a T-DNA inserted in exon 5 that results in reduced accumulation of a truncated pex5-10 protein [13]. The 35S:PEX5 construct used a PEX5 cDNA [17]; for 35S:PEX5 genotyping, PEX5 primers spanning introns were used to amplify larger genomic PEX5 or the smaller PEX5 cDNA products. PEX7 lacks introns, so a primer from the 35S promoter and a reverse PEX7 primer were used to genotype PEX7-overexpressing lines. (PDF 71 kb)