Green fluorescent proteins with short half-lives as reporters in Dictyostelium discoideum |
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Authors: | Heike Deichsel Shirin Friedel Anne Detterbeck Catharine Coyne Ursula Hamker H. K. MacWilliams |
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Affiliation: | Zoologisches Institut, Ludwig-Maximilians-Universit?t, Luisenstrasse 14, D-80333 München, Germany, DE
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Abstract: | We describe two modifications of the popular reporter green fluorescent protein (GFP) which have short half-lives in our system, the cellular slime mould Dictyostelium discoideum. One of these bears an N-terminal ubiquitin; this GFP was originally planned to be a substrate of the ”N-end-rule” pathway, but deubiquitination does not seem to occur, and a degradation by the UFD (ubiquitin-fusion-degradation pathway seems more probable. The protein half-life is about 3–5 h. The second construct has an N-terminus derived from the L11 ribosomal protein; it is transported to the nucleus and broken down much more rapidly than the ubiquitin fusion (protein half-life about 30 min). We show examples of the use of these reporters in the study of gene expression in Dictyostelium. Received: 20 April 1998 / Accepted: 23 August 1998 |
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