Isolation and characterization of the glucose-6-phosphate dehydrogenase gene of Drosophila melanogaster

Nucleotide sequence of the Drosophila glucose-6-phosphate dehydrogenase gene and comparison with the homologous human gene

“…We find this particularly puzzling since the restriction map reported by Hori et al (1985) is identical to that previously published by Ganguly et al (1985). Although we have no explanation for this discrepancy, it seems likely that the data presented in Fig.…”

“…Direct comparison of the genomic and cDNA sequences confirms the position of intron I but also indicates the presence of two additional small introns in the previously designated exon II (Ganguly et al, 1985). Therefore, exon II must now be redefined as containing three exons and two introns.…”

“…The purified fragment was denatured and hybridized with 10 ,ug of adult Drosophila poly(A) + RNA for 16 h at 46' C in a IO-$ reaction mixture. As described previously (Casey and Davidson, 1977;Ganguly et al, 1985), these conditions allow RNA/DNA hybridization in the absence of DNA/DNA hyb~d~ation. Following hybridization, the RNA/DNA hybrids were treated with S 1 nuclease and the size of the protected DNA fragment was resolved on a sequencing gel.…”

“…Following hybridization, the RNA/DNA hybrids were treated with S 1 nuclease and the size of the protected DNA fragment was resolved on a sequencing gel. A 6.7-kb HpaI-SstI DNA fragment (Ganguly et al, 1985) putatively containing the entire G6PD gene sequence was inserted into the single S&I site of the transformation vector Carnegie-20 (Rubin and Spradling, 1982) following conversion of the HpaI and SstI sites into XhoI sites. First, the 4.9-kb HindIII-BumHI fragment containing exons II-IV was cloned into pBR322.…”

“…G6PD activity has been used extensively for molecular, developmental, physiological and population studies with a variety of organisms, including Drosophila and man (Yoshida and Beutler, 1986). Recently, the gene for human and Drosophila G6PD have been cloned Martini et al, 1986;Takizawa et al, 1986;Gang&y et al, 1985;Hori et al, 1985). The amino acid sequence of human G6PD has been determined by direct amino acid sequence analysis of the purified enzyme (Takizawa et al, 1986), and it has been derived from the sequence of cDNA clones which encode the enzyme M.G.…”