Isolation of a structural gene mapping to subregions 63F of Drosophila melanogaster and 90B of D. hydei polytene chromosomes.

We have isolated by molecular cloning techniques a structural gene that maps to subregion 63F of Drosophila melanogaster chromosome 3L. This locus being one of the early chromosomal targets for ecdysone stimulation, may be induced to puff by the hormone. The gene is on a plasmid vector and it has been designated as pDm 63F. This recombinant molecule also maps to the early ecdysone inducible subregion 90B of Drosophila hydei chromosome 4. The cytological inspection of large number of chromosomal sets after in situ hybridization of the cloned DNA, locates the cloned sequence between bands 63F 2-4 according to Bridges map. Similarly, in Drosophila hydei the cloned DNA maps between subdivisions 90B 2-4 according to Berendes' map. In situ hybridization of the pDm 63F cloned DNA, directed to the nascent RNA rather than to the DNA, shows two to three times more silver grains over the corresponding regions when puffed than in the resting stage. There are however quantitative differences attending to the transcriptional activity of homologous loci in both species. By RNA excess hybridization we have found that the cellular concentration of the 63F cloned mRNA is bout 2 times higher in hormone stimulated total larval tissues than in non-stimulated ones.