HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Bingle, L. E. H. Articles by Gibson, W. C. Search for Related Content PubMed PubMed Citation Articles by Bingle, L. E. H. Articles by Gibson, W. C. Agricola Articles by Bingle, L. E. H. Articles by Gibson, W. C.

A novel GFP approach for the analysis of genetic exchange in trypanosomes allowing the in situ detection of mating events

School of Biological Sciences, University of Bristol, Bristol BS8 1UG, UK¹
Department of Clinical Veterinary Science, University of Bristol, Langford, Bristol BS40 7DU, UK²

Author for correspondence: Wendy C. Gibson. Tel: +44 117 928 8249. Fax: +44 117 925 7374. e-mail: w.gibson{at}bristol.ac.uk

Trypanosoma brucei undergoes genetic exchange in its insect vector by an unknown mechanism. To visualize the production of hybrids in the fly, a tetracycline (Tet)-inducible expression system was adapted. One parental trypanosome clone was transfected with the gene encoding Green Fluorescent Protein (GFP) under control of the Tet repressor in trans; transfection with these constructs also introduced genes for resistance to hygromycin and phleomycin, respectively. An experimental cross with a second parental clone carrying a gene for geneticin resistance produced fluorescent hybrids with both hygromycin and geneticin resistance. These results are consistent with the meiotic segregation and reassortment of the GFP and repressor genes. Fluorescent hybrids were visible in the salivary glands of the fly, but not the midgut, confirming that genetic exchange occurs among the trypanosome life cycle stages present in (or possibly en route to) the salivary glands. In conclusion, the experimental design has successfully produced fluorescent hybrids which can be observed directly in the salivary glands of the fly, and it has been shown that the recombinant genotypes were most probably the result of meiosis.

Keywords: Trypanosoma brucei, hybrids, meiosis, Green Fluorescent Protein, tsetse fly

Abbreviations: CM, Cunningham’s medium; GFP, Green Fluorescent Protein; UTR, untranslated region

^a Present address: School of Biological Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.

This article has been cited by other articles:

				T. Kulikowicz and T. A. Shapiro Distinct Genes Encode Type II Topoisomerases for the Nucleus and Mitochondrion in the Protozoan Parasite Trypanosoma brucei J. Biol. Chem., February 10, 2006; 281(6): 3048 - 3056. [Abstract] [Full Text] [PDF]

				M. A Miles, M D. Feliciangeli, and A. R. de Arias American trypanosomiasis (Chagas' disease) and the role of molecular epidemiology in guiding control strategies BMJ, June 25, 2003; 326(7404): 1444 - 1448. [Full Text] [PDF]