AN ANTIGENIC PEPTIDE OF MYOSIN HEAVY CHAIN-LIKE PROTEIN FROM TRICHINELLA SPIRALIS

Abstract

We produced antigenic peptide of Trichinella spiralis newborn larvae (NBL) which seemed to be a part (approximately one fourth) of myosin heavy chain, and some basic profiles were revealed. The cDNA library was constructed from NBL and immunoscreened with an antibody against the parasite. A clone, designated NBL21, was selected. It contained a cDNA transcript of 1656 by in length, which encoded 552-amino acids (64868 Da in the estimated molecular weight). The fusion protein encoded by the clone NBL21 was produced in an Escherichia coli expression system and affinity purified. NBL21 fusion proteins migrated at 64 kDa and reacted to T spiralis infected mouse sera and the antibody against NBL crude antigen. Antisera were developed against NBL21 fusion proteins, which reacted to a single band migrating at 200 kDa on Western blotting analysis of crude extracts from muscle larvae, and reacted to hypodermal muscles of T spiralis on immunohistochemical staining. The antigen was recognized by the mouse serum obtained from the early phase of infection, but the antigenicity was devoid of species specificity.