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Translation products of mRNA from infective larvae of Trichinella spiralis include an antigenic polypeptide

Published online by Cambridge University Press:  05 June 2009

Kazuo Sugane ,
Tadashi Matsuura  and
Hideaki Maekawa
Kazuo Sugane
Affiliation:
Department of Parasitology, School of Medicine, Shinshu University, Asahi 3-1-1, Matsumoto City, Japan
Tadashi Matsuura
Affiliation:
Department of Biology, Faculty of Science, Tokyo Metropolitan University, Fukazawa 2-1-1, Setagaya-ku, Tokyo, Japan
Hideaki Maekawa
Affiliation:
National Institute of Health, Kamioosaki 2-10-35, Shinagawa-ku, Tokyo, Japan
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Abstract

Total RNA was extracted from packed infective larvae of Trichinella spiralis by centrifugation through a 5·7 M caesium chloride cushion. Polyadenylated messenger RNA was separated from total RNA in an oligothymidylic acid-cellulose gel column. The in vitro translation of the mRNA, isolated from infective larvae of T. spiralis, was carried out using the rabbit reticulocyte cell-free translation system. Incorporation of 35S-methionine into trichloracetic acid precipitates in the lysate containing mRNA was 5 times greater than that in control. The translation products were analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by autoradiography. Many polypeptides with molecular weights of less than 100 000 were synthesized in the lysate. A T. spiralis positive mouse serum was mixed with translation products to form antigen-antibody complexes, which were then absorbed by Staphylococcus aureus Cowan 1 strain and analysed by autoradiography of SDS-PAGE. An antigenic polypeptide with a molecular weight of 48 000 was demonstrated to react specifically with IgG antibody in T. spiralis positive mouse serum. T. spiralis larvae were cultured in methionine-free medium containing 35S-methionine, and antigenic polypeptides in somatic extracts and ES products were compared with those in translation products by autoradiography of SDS-PAGE. Several polypeptides in ES products and somatic extracts reacted specifically with IgG anitbodies in positive serum. Especially the polypeptide with a molecular weight of 48 000 in ES products strongly reacted with IgG antibody in positive serum.

Keywords

Trichinella spiralismRNA translationantigens
Type
Research Papers
Information
Journal of Helminthology , Volume 61 , Issue 1 , March 1987 , pp. 1 - 8
Copyright
Copyright © Cambridge University Press 1987

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