Cloning and characterization of the gene encoding Schistosoma mansoni glutathione peroxidase

• Published in: Gene Vol. 138; no. 1; pp. 149 - 152
• Main Authors: Roche, Catherine, Williams, David L., Khalife, Jamal, LePresle, Thérése, Capron, André, Pierce, Raymond J.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 28.01.1994; Amsterdam Elsevier; New York, NY
• Subjects: Amino Acid Sequence; Animals; antioxidant enzyme; Base Sequence; Biological and medical sciences; Blotting, Southern; Cloning, Molecular; DNA - analysis; DNA - genetics; DNA, Protozoan; free radicals; Fundamental and applied biological sciences. Psychology; Genes, Helminth; Genes. Genome; Glutathione Peroxidase - genetics; helminth; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Parasite; Polymerase Chain Reaction; regulatory sequences; Schistosoma mansoni; Schistosoma mansoni - enzymology; Schistosoma mansoni - genetics; selenoprotein; UGA; vaccine; very small intron; aa; free radicals; SSC; SDS; nt; Parasite; GST; UGA; bp; Gpx; antioxidant enzyme; selenoprotein; regulatory sequences; vaccine; UTR; S; kb; RACE; helminth; very small intron; PCR; Glutathione peroxidase; Schistosoma mansoni; Nucleotide sequence; Enzyme; Transcription promoter; Plathelmintha; Gene organization; Trematoda; Peroxidases; Gene; Helmintha; Regulatory sequence; Oxidoreductases; Invertebrata; Molecular cloning; Structural analysis
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(94)90798-6

Antioxidant enzymes are thought to play a crucial role in the survival of the parasite, Schistosoma mansoni, during its migration through the tissues of the definitive host. We recently cloned the cDNA encoding one such enzyme, glutathione peroxidase (Gpx). In order to elucidate the regulation of expression of this gene, we describe the cloning and characterization of a Gpx gene of S. mansoni. An initial screen of a λEMBL4 genomic library using the corresponding cDNA sequence as a probe yielded 14 positive clones, two of which have so far been analyzed in detail. The complete Gpx gene contains five introns, four of which, located at the 5′ end, are extremely short (30–51 bp) and the last of which is approximately 6 kb long. We present the sequence of the gene including 73 bp at the 5′ end, the complete sequence to 137 bp downstream from the penultimate exon, 164 bp upstream and 131 bp downstream from the last 3′ exon. The potential mRNA cap site is situated 219 bp upstream from the ATG start codon. All intron/exon junctions correspond to the conventional eukaryotic splice signal. Analysis of the 5′ flanking region revealed the presence of a potential TATA box at −26 bp from the cap site, but no CAAT-like element is present. Southern blot analysis showed a unique Gpx gene organisation in the S. mansoni genome.