title: Isolation and characterization of Cu/Zn-superoxide dismutase in Fasciola gigantica.
abstract: A full-length complementary DNA (cDNA) encoding Cu/Zn-superoxide dismutase was isolated from Fasciola gigantica that on nucleotide sequencing showed a close homology (98.9%) with Cu/Zn-superoxide dismutase (SOD) of the temperate liver fluke, F. hepatica. Expression of the gene was found in all the three developmental stages of the parasite viz. adult, newly excysted juvenile and metacercaria at transcriptional level by reverse transcription-polymerase chain reaction (RT-PCR) and at the protein level by Western blotting. F. gigantica Cu/Zn-SOD cDNA was cloned and expressed in Escherichia coli. Enzyme activity of the recombinant protein was determined by nitroblue tetrazolium (NBT)-polyacrylamide gel electrophoresis (PAGE) and this activity was inactivated by hydrogen peroxide but not by sodium azide, indicating that the recombinant protein is Cu/Zn-SOD. The enzyme activity was relatively stable at a broad pH range of pH 4.0-10.0. Native Cu/Zn-superoxide dismutase protein was detected in the somatic extract and excretory-secretory products of the adult F. gigantica by Western blotting. NBT-PAGE showed a single Cu/Zn-SOD present in the somatic extract while three SODs are released ex vivo by the adult parasite. The recombinant superoxide dismutase did not react with the serum from buffaloes infected with F. gigantica. The role of this enzyme in defense by the parasite against the host reactive oxygen species is discussed.
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: H
authors: H
authors: Lalrinkima
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India. Electronic address: rainaok@rediffmail.com.
authors: O K
authors: OK
authors: Raina
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: Dinesh
authors: D
authors: Chandra
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: Siju Susan
authors: SS
authors: Jacob
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: R K
authors: RK
authors: Bauri
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: Subhash
authors: S
authors: Chandra
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: H S
authors: HS
authors: Yadav
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: M N
authors: MN
authors: Singh
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: A
authors: A
authors: Rialch
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: A
authors: A
authors: Varghese
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: P S
authors: PS
authors: Banerjee
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: Navneet
authors: N
authors: Kaur
authors: Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243122, U.P., India.
authors: Arvind
authors: A
authors: Sharma
mesh_terms: D000003:Abattoirs; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002020:Buffaloes / Q000469:parasitology; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification*; D018509:DNA, Helminth / Q000737:chemistry / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005209:Fasciola / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D005210:Fasciola hepatica / Q000201:enzymology / Q000235:genetics; D005211:Fascioliasis / Q000469:parasitology / Q000662:veterinary; D015971:Gene Expression Regulation, Enzymologic; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D008018:Life Cycle Stages / Q000235:genetics; D009580:Nitroblue Tetrazolium; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry; D017422:Sequence Analysis, DNA; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D018076:DNA, Complementary; D018509:DNA, Helminth; D007202:Indicators and Reagents; D018510:RNA, Helminth; D011994:Recombinant Proteins; D009580:Nitroblue Tetrazolium; D013482:Superoxide Dismutase
keywords: Cu/Zn–superoxide dismutase; Fasciola gigantica; Hydrogen peroxide; NBT–PAGE; Recombinant SOD; Sodium azide; Western blot