G. Mickey et al., USE OF DENATURING GRADIENT ELECTROPHORESIS TO DETERMINE THE DISTRIBUTIONS OF POLYMORPHISMS IN ENTIRE GENES IN NATURAL-POPULATIONS, Biochimica et biophysica acta, N. Gene structure and expression, 1260(2), 1995, pp. 123-131
We show here that the method of genomic denaturing gradient electropho
resis (gDGGE) can be used to examine any gene that has been previously
cloned and sequenced, and to detect and approximately localize within
the gene the majority of its polymorphisms. By using pooled-DNA gDGGE
, many different samples can be scanned on a single gel. Further, the
patterns on the gels and results from sequencing of some of the allele
s shows that a variety of different kinds of polymorphism, ranging fro
m single base changes to more substantial allelic differences, can be
distinguished. The extramacrochaetae (emc) gene of Drosophila melanoga
ster exhibits no polymorphism in its ORF that can be detected by this
method, although a rearrangement polymorphism was detected in the 3' d
ownstream region of the gene. The suppressor-of-hairless (Su(H)) gene
of D. melanogaster, however, exhibits a variety of polymorphisms in it
s ORF. Some are small deletions or insertions in a glutamine-rich part
of the gene product that would not have been detectable by ordinary s
creening methods. Many of the polymorphisms detected in this prelimina
ry survey are likely to have an impact on the function of the Su(H) ge
ne product.