Abstract
Glucose-6-phosphate dehydrogenase (G6PD) has a major role in NADPH production and is found in almost all cell types. The structural gene for G6PD is X-linked in Drosophila melanogaster, as it is in most eukaryotic organisms, and due to its ubiquitous expression, it can be considered a typical ‘housekeeping’ gene. Here we present the complete nucleotide (nt) sequence of G6PD cDNAs as well as the genomic copy of the G6PD gene. The G6PD gene has three introns so that the protein-coding region is divided into four segments. The 5'-end of mature G6PD mRNA is located 289 ± 1 nt upstream from the start codon. The sequence upstream from the transcription start point is G + T-rich and contains no commonly found transcription regulatory elements, such as a TATA box or GGGCGG sequence. D. melanogaster G6PD is 65 % homologous with the human G6PD protein but has no homology with the human sequence for the first 42 amino acid residues. The G6PD gene was shown to be active when transduced to autosomal positions. For each transformant, G6PD activity in both male and female adults was not significantly different, indicating that the transduced gene, unlike the resident G6PD, is not dosage-compensated in males.
Highlights
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
G6PD activity was measured as the increase in absorbance at 340 nm resulting from the reduction of NADP (Lucchesi and Rawls, 1973)
600 000 I recombinant phages from a cDNA library constructed from adult poly(A) + RNA were screened with DNA fragments containing exon I and exons II-IV of the genomic G6PD gene (Fig. 1)
Summary
Glucosed-phosphate dehydrogenase (G6PD) has a major role in NADPH production and is found in almost all cell types. We present the complete nucleotide (nt) sequence of G6PD cDNAs as well as the genomic copy of the G6PD gene. The G6PD gene has three introns so that the protein-coding region is divided into four segments. The 5’-end of mature G6PD mRNA is located 289 + 1 nt upstream from the start codon. D. melanogaster G6PD is 65% homologous with the human G6PD protein but has no homology with the human sequence for the first 42 amino acid residues. The G6PD gene was shown to be active when transduced to autosomal positions. G6PD activity in both male and female adults was not significantly different, indicating that the transduced gene, unlike the resident G6PD, is not dosage-compensated in males
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