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Science 9 June 1989:
Vol. 244. no. 4909, pp. 1167 - 1169
DOI: 10.1126/science.2499047
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Articles

Science, Vol 244, Issue 4909, 1167-1169
Copyright © 1989 by American Association for the Advancement of Science

articles

Expression of a bacterial gene in a trypanosomatid protozoan

V Bellofatto and GA Cross

Laboratory of Molecular Parasitology, Rockefeller University, New York, NY 10021.

A simple and reproducible assay for DNA-mediated transfection in the trypanosomatid protozoan Leptomonas seymouri has been developed. The assay is based on expression of the Escherichia coli chloramphenicol acetyl transferase (CAT) gene flanked by Leptomonas DNA fragments that are likely to contain necessary elements for gene expression in trypanosomes. After electroporation of cells in the presence of plasmid DNA, CAT activity was detected in crude cell lysates. No activity was detected when the orientation of the L. seymouri mini-exon sequence (placed upstream of the CAT gene) was reversed, or in additional control experiments. This system provides a method for defining transcriptional control elements in trypanosomes.


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