title: Molecular and immunological characterization of encoding gene and 14-3-3 protein 1 in Fasciola gigantica.
abstract: A cDNA encoding Fg14-3-3 protein 1 was cloned by immunoscreening of an adult-stage Fasciola gigantica cDNA library using a rabbit antiserum against tegumental antigens of the parasite. The protein has a deduced amino acid sequence of 252 residues and a calculated molecular weight of 28.7 kDa. It shows sequence identity values between 57.6 and 58.1% to the human 14-3-3 beta, zeta, theta, and eta proteins and is in a phylogenetic cluster with the 14-3-3 protein 1 of Schistosoma spp. Nucleic acid analyses indicate that the Fg14-3-3 protein 1 is encoded by a single copy gene and that this gene is expressed as a transcript of 1250 nucleotides. In adult and 4-week-old parasites the gene's transcriptional and translational products were localized in the gut epithelium, parenchyma, tegument cells, and in the reproductive organs. An antiserum against recombinant Fg14-3-3 protein 1 detected a slightly smaller 14-3-3 protein in the parasite's excretion/secretion material and showed cross-reactivity with 14-3-3 proteins in extracts of other trematodes and mouse. Antibodies against Fg14-3-3 protein were detected in the sera of rabbits as early as 2 weeks after infection with metacercariae of F. gigantica and the antibody titre increased continuously over a 10-week observation period.
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, 10400, Thailand.
authors.forename: K
authors.initials: K
authors.lastname: Chaithirayanon
authors.forename: R
authors.initials: R
authors.lastname: Grams
authors.forename: S
authors.initials: S
authors.lastname: Vichasri-Grams
authors.forename: A
authors.initials: A
authors.lastname: Hofmann
authors.forename: G
authors.initials: G
authors.lastname: Korge
authors.forename: V
authors.initials: V
authors.lastname: Viyanant
authors.forename: E S
authors.initials: ES
authors.lastname: Upatham
authors.forename: P
authors.initials: P
authors.lastname: Sobhon
mesh_terms: D048948:14-3-3 Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood / Q000276:immunology; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D005209:Fasciola / Q000235:genetics* / Q000254:growth & development / Q000276:immunology / Q000378:metabolism; D005211:Fascioliasis / Q000276:immunology*; D005260:Female; D015723:Gene Library; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017422:Sequence Analysis, DNA
chemical_list: D048948:14-3-3 Proteins; D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018510:RNA, Helminth; D011994:Recombinant Proteins