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. 2007 Jun;144(2):821–835. doi: 10.1104/pp.107.096214

Figure 3.

Figure 3.

Identification of a Mu insertion in the maize Zmsmu2 gene that is tightly linked with the opaque kernel phenotype. A, Mu TAIL-PCR products from genomic DNA of seedlings of opaque F2 kernels (lanes 1–6), seedlings from vitreous F2 kernels (lanes 7–12), and homozygous wild-type W64A+ seedlings (lanes 13–18). A 150-bp fragment (arrow) was detected in DNA of all the opaque mutants (lanes 1–6) but none of the homozygous wild types (lanes 13–18). This DNA was detected in some of the vitreous F2 progeny (lanes 9–11), which may be heterozygous for zmsmu2-1. B, Linkage analysis of zmsmu2-1 with the opaque kernel phenotype. Genomic PCR using gene-specific primers F3 and R6 (lanes with a plus sign; see C and D for the position of the primers in the Zmsmu2 gene) yielded a DNA band specific for the wild-type allele at the Zmsmu2 locus, whereas another PCR using primers MuTir6 and R6 (lanes with a minus sign) produced a zmsmu2-1-specific band. The lane numbers correspond to F2 individuals that provided genomic DNA templates for the PCR. C, Structure of the maize Zmsmu2 gene. White and black boxes correspond to exon sequences in the 5′ and 3′ UTRs and the coding region, respectively, and gray bars between the boxes represent introns. The Mu element (not to scale) in alleles zmsmu2-1 to zmsmu2-4 is illustrated by reverse triangles; the allele numbers are specified in the black triangles. Black and gray arrowheads mark the positions of gene-specific primers and of a Mu-specific primer, MuTir6, respectively, used for PCR amplification. D, Alignment of the Zmsmu2 genomic DNA sequence from two maize inbreds, W64A+ and B73+. Only the 5′ ends of the available sequences are shown. The nucleotide sequence in gray corresponds to the Mu-containing sequence obtained from zmsmu2-1 cDNA. Transcription may start around the F1 primer site, because we were able to detect Zmsmu2 cDNA from wild-type RT products using the F1 and R5 primer pair (see Fig. 5A). Intron sequences are italicized. Capital letters over the W64A+ sequence are the first 16 amino acid residues of the deduced protein. The three sequences in boxes show the target duplication sites for Mu insertion in zmsmu2-1, zmsmu2-3, and zmsmu2-4. Primer sequences are shown in arrows. Underlined nucleotide sequences are conserved cis-elements identified by Transcription Element Search System (http://www.cbil.upenn.edu/tess).