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. Author manuscript; available in PMC: 2010 May 11.
Published in final edited form as: Curr Biol. 2004 Mar 9;14(5):412–417. doi: 10.1016/j.cub.2004.02.032

Figure 3. Map-Based Cloning and Gene and Protein structure of AtMAP65-3/PLE.

Figure 3

(A) Genetic map of the PLE region. Distances between molecular markers represent absolute numbers of recombinants. (B) Overlapping cosmid clones used for complementing ple mutants aligned with the genetic map in (A). Clone N16 (open box) complemented ple alleles, whereas the clones represented by the shaded boxes did not. The genomic DNA of clone N16 codes for three full and two partial open reading frames, and these are indicated by the arrows. Allele specific polymorphisms were only detected in the At5g51600 gene, which is AtMAP65-3. (C) The AtMAP65-3/PLE gene. There are 11 introns, with one in the 5′ UTR. The open reading frame encodes a 707 amino acid protein. Mutations in ple-5 and ple-6 introduce stop codons. The ple-1 mutation affects the splicing efficiency of intron II. (D) RT-PCR of cDNA from ple-1 (ple-1c) and wild- type cDNA (Col-c) showing the aberrant splicing of intron II. The majority of transcripts are not spliced and would result in a protein of 61 amino acids. M, markers; Col-g, genomic DNA control using the same primers; W, no template control. (E) Aligned wild-type and ple-1 sequences showing the mutation (arrowed) causing the aberrant splicing.