Green Fluorescent Protein As a Cell-Labeling Tool and a Reporter of Gene Expression in Transgenic Rainbow Trout |
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Authors: | Yutaka Takeuchi Goro Yoshizaki Toshio Takeuchi |
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Institution: | (1) Laboratory of Fish-Culture, Tokyo University of Fisheries, Konan, Minato-ku, Tokyo 108-0075, Japan, JP |
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Abstract: | Green fluorescent protein (GFP) has been used as an indicator of transgene expression in living cells and organisms. For
testing the utility of GFP in rainbow trout, we microinjected fertilized eggs with four types of supercoiled constructs containing
two variants of GFP complementary DNA (S65T and EGFP), driven by two ubiquitous regulatory elements, human cytomegalovirus
immediate early enhancer-promoter (CMV) and Xenopus laevis elongation factor 1α enhancer-promoter (EF1).
Green fluorescence was first observed at 3 days postfertilization, when the embryo was in the mid-blastula stage. Fluorescence
could be detected mosaically in various types of embryonic cells and tissues of swim-up fry. Both the percentage of fluorescent
cells and the fluorescence intensity of GFP-expressing cells on blastoderms, measured with a microscopic photometry system,
were highest in CMV-EGFP-microinjected embryos. We conclude that GFP is capable of producing detectable fluorescence in rainbow
trout, and can be a powerful tool as a cell marker and reporter gene for cold-water fish, and that analysis of GFP expression
in living cells is useful for characterizing the activity of cis-elements in vivo.
Received December 21, 1998; accepted March 31, 1999. |
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Keywords: | : GFP microinjection rainbow trout transgenic fish gene expression gene transfer |
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