[Prokaryotic expression of an antigen gene of Trichinella spiralis and identification of the recombinant protein].
OBJECTIVE
To obtain the recombinant protein of an antigen gene Ts88 of Trichinella spiralis and identify the characteristics of the recombinant protein.


METHODS
Ts88 cDNA obtained by immunoscreening the cDNA library of adult T. spiralis was subcloned into the pET-28c(+) expression vector and expressed in E. coli. Mice were immunized with the fusion protein incorporated into Freund' s adjuvant and the immune sera were collected. The titers of the Ts88 immune sera and the antigenicity of the recombinant protein were detected by ELISA and Western blotting. Immuno-fluorescence test was performed in order to confirm the distribution of Ts88 protein in the worm.


RESULTS
The fragment of Ts88 gene was expressed successfully in E. coli and a highly purified fusion protein was obtained. Immunization with the recombinant protein in mice produced high titers of antibodies, which recognized some components of native antigens of soluble proteins from adult worm of T. spiralis. Western blotting analysis showed that Ts88 recombinant antigen was recognized by all the positive sera, such as the sera from infected or immunized rabbits, from infected swine and from patients of trichinosis. Immuno-fluorescence test confirmed that Ts88 protein mainly distributed in the cuticle surface of the worm.


CONCLUSIONS
The Ts88 antigen gene from T. spiralis was successfully expressed. The recombinant protein presented antigenicity.
Lei
Li-ping
LP
Department of Parasitology, Capital University of Medical Sciences, Beijing 100054, China.
Zhu
Xin-ping
XP
Yang
Jing
J
Yang
Ya-ping
YP
Ding
Li
L
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000096:biosynthesis* / Q000276:immunology* / Q000302:isolation & purification; D005260:Female; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology* / Q000302:isolation & purification; D017160:Trichinella spiralis / Q000276:immunology*
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D007106:Immune Sera; D011994:Recombinant Proteins