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Molecular analysis of the gene encoding an antigenic polypeptide of Trichinella spiralis infective larvae

Published online by Cambridge University Press:  05 June 2009

Kazuo Sugane  and
Tadashi Matsuura
Kazuo Sugane
Affiliation:
Department of Parasitology, Faculty of Medicine, Shinshu University, Asahi 3-1-1, Matsumoto City, Nagano Prefecture, Japan
Tadashi Matsuura
Affiliation:
Department of Parasitology, Faculty of Medicine, Shinshu University, Asahi 3-1-1, Matsumoto City, Nagano Prefecture, Japan
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Abstract

The gene encoding an antigenic polypeptide of Trichinella spiralis infective larvae was studied using recombinant DNA techniques. cDNA synthesized from poly(A)-rich mRNA from T. spiralis infective larvae was ligated into phage vector λgt11 DNA and packaged in vitro. The phages were propagated on Escherichia coli and a λgt11 expression library was constructed. A cDNA clone encoding a 46 kDa antigenic polypeptide was selected by immunoscreening of the library and identified by the epitope selection method. A clone containing nearly full-length cDNA for a 46 kDa protein was isolated. The gene encoding this 46 kDa antigenic polypeptide was characterized by DNA and RNA blot analysis using the cDNA as a probe. The gene was transcribed to mRNA with approximately 1400 nucleotides and translated to 46 kDa polypeptide. The antigenic polypeptide was excreted/secreted as a 46 kDa native antigen. The antigenic β-galactosidase fusion protein synthesized by bacteria had no cross-reactivity with other parasite-infected sera.

Keywords

Trichinella spiralisfusion proteinepitope selectionλgt11
Type
Research Article
Information
Journal of Helminthology , Volume 64 , Issue 1 , March 1990 , pp. 1 - 8
Copyright
Copyright © Cambridge University Press 1990

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