title: Cloning and analysis of a cDNA encoding a putative serine protease comprising two trypsin-like domains of Trichinella spiralis.
abstract: The cDNA encoding a putative serine protease, TsSerP, was cloned by degenerative polymerase chain reaction and screening of the cDNA library from Trichinella spiralis adult-newborn larvae stage. Sequence analysis revealed the presence of two trypsin-like serine protease domains flanking a hydrophilic domain, with the catalytic triad residue histidine in the alpha domain substituted by an arginine residue. Southern blots indicated that this was a single copy gene in the parasite genome. Northern blots demonstrated a single 2.3-kb transcript during the muscle larvae and adult stages of T. spiralis. The recombinant protein from the TsSerP beta domain (betaSerP) was produced but not recognised by T. spiralis-infected swine serum. An anti-betaSerP polyclonal serum detected a 69-kDa polypeptide in the soluble antigens of T. spiralis muscle larvae. Immunolocalisation analysis located TsSerP on the inner layer of the cuticle and oesophagus of the parasite, suggesting a potential role in its moulting and/or digestive functions.
authors.affiliation: UMR 956 BIPAR (INRA-AFSSA-ENVA-UPVM), 23 avenue du Général de Gaulle, 94703, Maisons-Alfort, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Trap
authors.forename: Baoquan
authors.initials: B
authors.lastname: Fu
authors.forename: Franck
authors.initials: F
authors.lastname: Le Guerhier
authors.forename: Mingyuan
authors.initials: M
authors.lastname: Liu
authors.forename: Danielle
authors.initials: D
authors.lastname: Le Rhun
authors.forename: Thibault
authors.initials: T
authors.lastname: Romand
authors.forename: Catherine
authors.initials: C
authors.lastname: Perret
authors.forename: Radu
authors.initials: R
authors.lastname: Blaga
authors.forename: Pascal
authors.initials: P
authors.lastname: Boireau
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D018509:DNA, Helminth / Q000032:analysis*; D015723:Gene Library; D017238:Genes, Helminth; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D017160:Trichinella spiralis / Q000201:enzymology* / Q000235:genetics; D014357:Trypsin / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D018509:DNA, Helminth; D012697:Serine Endopeptidases; D014357:Trypsin