Molecular and immunological characterization of encoding gene and 14-3-3 protein 1 in Fasciola gigantica.
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.
Chaithirayanon
K
K
Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, 10400, Thailand.
Grams
R
R
Vichasri-Grams
S
S
Hofmann
A
A
Korge
G
G
Viyanant
V
V
Upatham
E S
ES
Sobhon
P
P
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
D048948:14-3-3 Proteins; D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018510:RNA, Helminth; D011994:Recombinant Proteins