Figure 9.

Schematic Representation of the Mutant Allele wp Genomic Sequence and Details of the Tmg-Express1 Insertion in Intron II.

(A) Genomic sequence of the mutant wp allele obtained from PCR fragments amplified from the wp line 2 (Table 1). The introns are indicated and their length given in base pairs. The 5725-bp Tgm-Express1 insertion in Intron II, 231 bp into the intron, is drawn at the top as a double line, with the arrowheads representing the inverted repeats. The full length of the mutant gene, 9251 bp, is written to the right of the thicker solid line representing the cDNA if proper splicing should take place.

(B) Detailed representation of the Tgm-Express1 insertion drawn to scale and flanked by the target site duplication, TGA. Arrowheads represent the 24-bp imperfect inverted repeats. The segments of sequence with high identity scores to soybean EST sequences in GenBank are indicated by the solid blocks and intron regions by gray boxes. The annotation, clone IDs, and GenBank accession numbers are given underneath. The first portion of the transposon insertion with 97% identity to a genomic sequence entered in GenBank as G. max CDC2 (1) pseudogene is measured at top with the 1697-bp line and accession number U64200. The dashed line pointing upstream of the 1697-bp portion of the U64200 genomic fragment represents the diverging sequence with no similarity to the F3H Intron II to the left of the insertion. Instead, it matches an EST with accession number CF922959. The left border inverted repeat of U64200 identical to that in the wp insertion is represented with an arrowhead. The sequence divergence upstream of the identical left border inverted repeats reveals the existence of a second Tgm-Express2 insertion somewhere else in the genome. CDC2, cell division cycle 2; FPKFB2, fructose-6-phosphate 2-kinase/fructose-2-6-biphosphatase; MDH, malate dehydrogenase; CS, Cys synthase.