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The Plant Cell, Vol. 4, 119-128,February 1992O 1992American Society of Plant Physiologists RESEARCH ARTICLE

Summary: The Plant Cell, Vol. 4, 119-128,February 1992O 1992American Society of Plant Physiologists
Cloning the Arabidopsis GA7 Locus by Genomic Subtraction
Tai-ping Sun,' Howard M. Goodman, and Frederick M. Ausube12
Department of Genetics, Harvard Medical School, and Department of Molecular Biology, MassachusettsGeneral Hospital,
Boston, Massachusetts02114
Arabidopsis thalianagal mutantsare gibberellin-responsivedwarfs. We usedthe genomic subtractiontechniqueto clone
DNA sequencesthat are presentin wild-type Arabidopsis(ecotypeLandsbergerecta, Ler) but are missing in a presump-
tive gal deletion mutant, gal-3. The cloned sequences correspondto a 5.0-kb deletion in the gal-3 genome. Three lines
of evidence indicatedthat the 5.0-kb deletion in the gal-3 mutant is locatedat the GAl locus. First, restriction fragment
length polymorphism mapping showed that DNA sequences within the 5.0-kb deletion map to the GAl locus. Second,
cosmid clones that contain wild-type DNA inserts spanning the deletion in gal-3 complemented the dwarf phenotype
when integrated into the gal-3 genome by Agrobacterium tumefaciens-mediated transformation. Third, we identified
molecular lesions in four additional gal alleles within the 5.0-kb region deleted in mutant gal-3. One of these lesions
is a large insertion or inversionlocated within the most distal intron encoded by the GAl locus. The three other lesions
are all single base changes locatedwithin the two most distal exons. RNA gel blot analysis indicatedthat the GAl locus
encodes a 2.8-kb mRNA. We calculated a recombination rate of 10-5 cM per nucleotide for the GAl region of the
Arabidopsis genome.
Cloninga plantgenethat isdefinedsolely bya mutantpheno-


Source: Ausubel, Frederick M. - Department of Genetics, Harvard University


Collections: Biology and Medicine