Figure 2.

Map-based cloning of HCF222 and gene expression in the mutants. A, Mapping of the hcf222-1 locus. The gene was mapped on the upper arm of chromosome 5 between the single-nucleotide polymorphism markers 483168 and 483268. Genes located in the region around the gene HCF222/At5g15802 are depicted. B, Gene structure of HCF222 and mutant alleles. The coding sequence of HCF222 comprises 943 nucleotides consisting of two exons separated by an intron at nucleotide positions 114 to 756. Sequence analysis of the mutant allele hcf222-1 revealed a point mutation at nucleotide 98 in exon 1. This mutation modifies codon GCT into GTT, causing an amino acid exchange from the original Ala to Val. In hcf222-2 (GABI-Kat line 038B09), the T-DNA insertion was detected in the intron between nucleotides 347 and 348. C, Determination of HCF222 mRNA levels in the wild type and the two allelic mutants by real-time PCR. n.d., Not detectable. Expression levels of HCF222 were determined in three biological replicates, and means with sd are depicted.