Cloning and analysis of a cDNA encoding a putative serine protease comprising two trypsin-like domains of Trichinella spiralis.
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.
Trap
Catherine
C
UMR 956 BIPAR (INRA-AFSSA-ENVA-UPVM), 23 avenue du Général de Gaulle, 94703, Maisons-Alfort, France.
Fu
Baoquan
B
Le Guerhier
Franck
F
Liu
Mingyuan
M
Le Rhun
Danielle
D
Romand
Thibault
T
Perret
Catherine
C
Blaga
Radu
R
Boireau
Pascal
P
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
D018076:DNA, Complementary; D018509:DNA, Helminth; D012697:Serine Endopeptidases; D014357:Trypsin