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Fig. 1. Map-based cloning and characterization of hve mutations.
(A) Strategy followed to identify the HVE gene. After
analyzing 3090 chromosomes, 41 recombinants (shown in parentheses) placed the
HVE gene in the interval flanked by the nga1145 and T17M13B markers.
Sequencing of segments of the candidate region in the informative recombinants
rendered five novel SNPs, which allowed us to map the HVE gene within
a 61 kb interval encompassed by the T8K22 BAC. (B-D) Structure of the
HVE gene of Arabidopsis with the position and nature of
hve mutations indicated. (B) Schematic representation of the
HVE gene. Exons and introns are depicted as black boxes and lines,
respectively. The predicted translation start (ATG) and stop (TAA) codons are
indicated. The region harboring the 16 nt deletion found in the hve-1
allele is aligned with those of the wild-type HVE alleles of Col-0
and Ws-2, the splicing donor signals of which are boxed. (C) PCR amplification
products encompassing the 14th exon and 14th intron of HVE, using as
templates genomic DNA (gDNA) and cDNA from Col-0, Ws-2 and
hve-1/hve-1 plants. The three different splicing products identified
in hve-1 cDNA are numbered. M, molecular weight marker; NC, negative
control. (D) Partial alignment of the predicted HVE proteins from Col-0 and
two of the splicing products of hve-1. Numbers correspond to amino
acid positions. Asterisks represent premature stop codons.
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