Figure 3.

AHAS621 conversion examined by restriction fragment length polymorphism and sequence analysis. (A) A map of the amplified AHAS621 target sequence from both wild-type and mutant alleles indicates the positions of PCR primers and the BfaI restriction site. (B) Polymorphism of wild-type and mutant alleles in target PCR fragments from positive control (P), negative control (N), and a representative event (E) before (U) and after BfaI restriction (R). (C) Sequence comparison of cloned AHAS621 alleles from the above samples. (D) Sequence comparison of cloned AHAS165 alleles from a positive control event (P), a negative control event (N), and two events with the predicted nucleotide conversion (E). Sequences were generated directly from PCR-amplified DNA from maize tissues. Because multiple AHAS genes exist in maize, as expected, both unconverted wild-type and converted mutant alleles are present in the events, as represented by the two overlapping peaks and the N nucleotide designation in the chromatograms.