Advantages and Limitations of Using Spm as a Transposon Tag
Transposon tagging has become the method of choice for isolating genes whose products are in low abundance. We have recently used the transposable element Spm to tag and clone maize regulatory loci. Our choice of Spm was dictated by several factors: The frequency of transposition of Spm is high enough to obtain detectable transposition events, into loci affecting kernel traits, in populations of <106 seed. Although the copy number of Spm is high in the maize genome, insertions into the gene of interest can be distinguished from other Spm copies by digesting DNAs from segregating populations with methyl-sensitive restriction enzymes, and hybridizing with Spm-specific probes. Since all members of the Spm family thus far examined share DNA homology, hybridization with appropriate probes allows detection of insertions of both autonomous and defective elements. Thus, if a mutable allele can be shown to be under Spm control, one can be reasonably confident of successfully cloning that allele.
KeywordsTransposable Element Mutable Allele Transposition Event Mapping Restriction Fragment Length Polymorphism Sensitive Restriction Enzyme
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