Figure 5.

Positional cloning and molecular characterization of FIN219. (A) Positional cloning of FIN219. Genetic mapping first located the gene on the lower arm of chromosome 2 between SSLP marker AthBio2 and CAPS marker 90J19T7. The arrow points to the telomere on the bottom arm of chromosome 2. First-round analysis of recombinant lines positioned the gene within the overlapping region of CIC10H6 and CIC5C4. Further mapping of the gene using overlapping BAC clones covering the region located the gene within a single BAC clone (TAM7G15). Overlapping binary cosmid clones were constructed using this BAC clone to transform fin219 mutants. Two cosmid clones (nos. 36 and 42) rescued the mutant phenotype. The overlapping region between clones 36 and 42 was dissected further into 5.1- and 7.2-kb BamHI fragments and used for functional rescue again. Only the 5.1-kb BamHI fragment complemented the mutant fin219. (B) BamHI; (H) HindIII; (Xb) XbaI. FIN219 is located between SPA1 and an unknown gene, F11C10.7. (B) Diagram of the 5.1-kb BamHI fragment, which contains one transcribed region with five predicted exons and four introns. The locations of the initiation codon (ATG) and stop codon are indicated by stars. (C) Sequence alignment of FIN219 (accession no. AF279129) and a GH3-like protein (accession no. AF071527.1) from Arabidopsis. Identical residues of these two proteins are shown in shaded boxes.