Molecular cloning and characterization of a superoxide dismutase (sod) gene in Pneumocystis carinii.
This work reports the isolation and characterization of a gene encoding a superoxide dismutase (SOD, EC.1.15.1.1.) from Pneumocystis carinii derived from rat. Sense and antisense oligonucleotides, deduced from SOD amino acid sequences from a wide variety of organisms, allowed amplification of a 669 bp genomic DNA fragment specific to this P. carinii. RACE-PCR was used to obtain the major part of the complementary DNA; the 5'- and 3'-genomic regions were obtained respectively from a Mbol subgenomic library and from an amplified fragment using oligonucleotides designed from the cDNA sequence. Comparison of genomic and cDNA sequences showed an open reading frame of 660 bp interrupted by seven small introns. The deduced amino acid sequence contained 220 residues. Protein sequence alignment demonstrated the highest homology (50.5% identity; 70.3% similarity) with Saccharomyces cerevisiae manganese-SOD (MnSOD) suggesting that P. carinii SOD belongs to the mitochondrial MnSOD group. A putative targeting peptide found at the 5'-end of the P. carinii SOD sequence also suggested its mitochondrial localization.
Denis
C M
CM
INSERM U42, Villeneuve d'Ascq, France. cdenis@worf.molbiol.ox.ac.uk
Guyot
K
K
Wakefield
A E
AE
Dive
D
D
Dei-Cas
E
E
Camus
D
D
Odberg-Ferragut
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000201:enzymology; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D008969:Molecular Sequence Data; D011010:Pneumocystis / Q000201:enzymology* / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D051381:Rats; D011916:Rats, Inbred F344; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*
D017931:DNA Primers; D013482:Superoxide Dismutase