Figure 2

Molecular genetics and transformation rescue of bas-1.(A) Genetic and physical map of bas-1 region and bas-1 mutant alleles. Locations and extent of mutations for each bas-1 allele are shown to scale with respect to C05D2.4 and C05D2.3 coding sequences, based on known splicing patterns or Genefinder predictions. Exons are indicated by rectangular bars; an alternatively spliced 27 bp exon is also indicated after exon 2 in C05D2.4. Four of five bas-1 mutants affect only C05D2.4; ad446 is a larger deletion removing most coding sequence of both C05D2.4 and C05D2.3. (B) Genomic DNA constructs that rescue or fail to rescue bas-1 mutants. Constructs are shown to scale (top), based on the 15.1 kbp insert of the plasmid clone C05D2XN; construct names are indicated in the box on the left. The cosmid C05D2 is larger, as indicated by the arrows. Clones below are subclones or modifications of C05D2XN. Coding regions for the two predicted AADC genes C05D2.4 and C05D2.3 are indicated by the blue boxes; intergenic regions are shown in yellow. C05D2XN upstream of C05D2.4 contains two other predicted genes, one complete (C05D2.8) and one partial (C05D2.5). There are no predicted genes in the 3 kb downstream of C05D2.3. In the constructs with the least upstream sequence, only a portion of C05D2.8 remains. Constructs mutated to introduce premature stop codons are indicated with a X in the coding sequence, and red downstream of the introduced stop codon. The construct pCL8001 has a GFP gene inserted in a manner that would inactivate the C05D2.3 gene, so is comparable to the pCL7991 construct. [No GFP expression was seen in the C05D2.3::GFP reporter construct lines.]