Cloning and differential expression of manganese superoxide dismutase (Mn-SOD) of Trichinella pseudospiralis.
The complete coding sequence of manganese superoxide dismutase (Mn-SOD) of Trichinella pseudospiralis (Tp) was obtained and characterized for the first time by degenerative reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of complementary DNA ends (RACE) reactions. The open reading frame of Tp Mn-SOD contained 663 nucleotides, encoding 220 amino acid residues. This included the conserved histidine and aspartate residues for metal binding, cysteine residues for disulfide bond formation, and arginine residue for directing the superoxide ion to the protein. The presence of mitochondrial transit peptides and maturation cleavage site suggest that the cloned Tp Mn-SOD gene is a mitochondrial enzyme. It is a single copy gene containing three introns. Northern blotting suggested that the expression level of Mn-SOD is lower than Cu/Zn SOD in infective stage larvae. Semi-quantitative RT-PCR demonstrated that a single dominant transcript of Tp Mn-SOD was highly expressed in the infective-stage larvae but not in adult worms. The information provides a better understanding of the highly compartmentalized superoxide dismutases of adenophorean nematodes.
Wu
W K
WK
Department of Zoology, The University of Hong Kong, Pokfulam, Hong Kong, SAR, People's Republic of China.
Mak
C H
CH
Ko
R C
RC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018509:DNA, Helminth / Q000032:analysis; D015971:Gene Expression Regulation, Enzymologic; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014234:Trichinella / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D013482:Superoxide Dismutase