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THE PLANT CELL, Vol 4, Issue 2 119-128, Copyright © 1992 by American Society of Plant Biologists
Cloning the Arabidopsis GA1 Locus by Genomic Subtraction
Tp. Sun, H. M. Goodman and F. M. Ausubel
Department of Genetics, Harvard Medical School, and Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts 02114
Arabidopsis thaliana ga1 mutants are gibberellin-responsive dwarfs. We used
the genomic subtraction technique to clone DNA sequences that are present
in wild-type Arabidopsis (ecotype Landsberg erecta, Ler) but are missing in
a presumptive ga1 deletion mutant, ga1-3. The cloned sequences correspond
to a 5.0-kb deletion in the ga1-3 genome. Three lines of evidence indicated
that the 5.0-kb deletion in the ga1-3 mutant is located at the GA1 locus.
First, restriction fragment length polymorphism mapping showed that DNA
sequences within the 5.0-kb deletion map to the GA1 locus. Second, cosmid
clones that contain wild-type DNA inserts spanning the deletion in ga1-3
complemented the dwarf phenotype when integrated into the ga1-3 genome by
Agrobacterium tumefaciens-mediated transformation. Third, we identified
molecular lesions in four additional ga1 alleles within the 5.0-kb region
deleted in mutant ga1-3. One of these lesions is a large insertion or
inversion located within the most distal intron encoded by the GA1 locus.
The three other lesions are all single base changes located within the two
most distal exons. RNA gel blot analysis indicated that the GA1 locus
encodes a 2.8-kb mRNA. We calculated a recombination rate of 10-5 cM per
nucleotide for the GA1 region of the Arabidopsis genome.
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