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DNA-binding activity in the excretory–secretory products of Trichinella pseudospiralis (Nematoda: Trichinelloidea)

Published online by Cambridge University Press:  28 November 2001

C. H. MAK  and
R. C. KO
C. H. MAK
Affiliation:
Department of Zoology, The University of Hong Kong, Hong Kong, China
R. C. KO
Affiliation:
Department of Zoology, The University of Hong Kong, Hong Kong, China
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Abstract

A novel DNA-binding peptide of Mr∼30 kDa was documented for the first time in the excretory–secretory (E–S) products of the infective-stage larvae of Trichinella pseudospiralis. Larvae recovered from muscles of infected mice were maintained for 48 h in DMEM medium. E–S products of worms extracted from the medium were analysed for DNA-binding activity by the electrophoretic mobility shift assay (EMSA). Multiple DNA-protein complexes were detected. A comparison of the Mr of proteins in the complexes indicated that they could bind to the target DNA as a dimer, tetramer or multiples of tetramers. Site selection and competition analysis showed that the binding has a low specificity. A (G/C-rich)-gap-(G/T-rich)-DNA sequence pattern was extracted from a pool of degenerate PCR fragments binding to the E–S products. Results of immunoprecipitation and electrophoretic mobility supershift assay confirmed the authenticity of the DNA-binding protein as an E–S product.

Keywords

Trichinella pseudospiralisDNA-binding proteinexcretory–secretory productsnematode
Type
Research Article
Information
Parasitology , Volume 123 , Issue 3 , March 2001 , pp. 301 - 308
Copyright
© 2002 Cambridge University Press

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