Cloning and dosage compensation of the 6-phosphogluconate dehydrogenase gene (Pgd+) of Drosophila melanogaster.
Using a heterologous rat cDNA probe, we have identified a 14.7 kbp Drosophila melanogaster genomic clone containing the X-linked gene Pgd+, which encodes the enzyme 6-phosphogluconate dehydrogenase (6PGD). We used in situ hybridization to larval polytene chromosomes, a somatic transient expression assay for enzyme activity, and the rescue of the lethal Pgd- phenotype by germline transformation to verify the identity of the gene. A 7.4 kbp fragment including the gene and approximately 1.2 kbp of upstream and 1.8 kbp of downstream sequences was relocated to autosomal ectopic sites by germline transformation; this transduced gene exhibits levels of enhanced activity in males comparable to those of the indigenous gene at its normal X chromosome locus. We conclude that the sequences responsible for dosage compensation of Pgd+ are included in this fragment.
Gutierrez
A G
AG
Department of Biology, University of North Carolina, Chapel Hill 27514.
Christensen
A C
AC
Manning
J E
JE
Lucchesi
J C
JC
D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004303:Dosage Compensation, Genetic; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics*; D005260:Female; D008040:Genetic Linkage; D008297:Male; D010734:Phosphogluconate Dehydrogenase / Q000235:genetics*; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014161:Transduction, Genetic; D014960:X Chromosome
D004247:DNA; D010734:Phosphogluconate Dehydrogenase