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Genomic cloning of human Echinococcus granulosus DNA: isolation of recombinant plasmids and their use as genetic markers in strain characterization

Published online by Cambridge University Press:  06 April 2009

A. K. Rishi  and
D. P. McManus
A. K. Rishi
Affiliation:
Department of Pure and Applied Biology, Imperial College of Science and Technology, Prince Consort Road, London SW7 2BB
D. P. McManus
Affiliation:
Department of Pure and Applied Biology, Imperial College of Science and Technology, Prince Consort Road, London SW7 2BB
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Summary

A small, size selected (0·5–5·0Kbp) genomic DNA library has been constructed in the bacterial plasmid pAT153 using DNA extracted from a human isolate (Kenyan origin) of the hydatid disease organism, Echinococcus granulosus. A panel of taeniid cestode DNAs has been used in conjunction with hybridization and restriction-enzyme analysis to identify in the library two recombinant plasmids with Echinococcus-specific inserts and a single recombinant plasmid (coded pEG18) with a DNA fragment unique for E. granulosus. These, and other recombinant plasmids with E. granulosus DNA inserts, have been used in restriction endonuclease, Southern transfer and hybridization analysis to independently and repro-ducibly discriminate between the UK horse and sheep strains of E. granulosus; these probes may well prove of value for characterizing isolates from other endemic areas. The feasibility of using a cloned DNA fragment as the basis of a simple field test for distinguishing eggs of E. granulosus from those of other taeniid cestodes is discussed. The recombinant insert (approximately 2·3 Kbp in size) of pEG18 has a low copy number - estimated at approximately 26 - and it may not be sufficiently sensitive for practical use as a DNA probe in the identification of small numbers of E. granulosus eggs by molecular hybridization.

Type
Research Article
Information
Parasitology , Volume 94 , Issue 2 , April 1987 , pp. 369 - 383
Copyright
Copyright © Cambridge University Press 1987

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