Generation of Two-color Transgenic Zebrafish Using the Green and Red Fluorescent Protein Reporter Genes gfp and rfp |
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Authors: | Haiyan Wan Jiangyan He Bensheng Ju Tie Yan Toong Jin Lam Zhiyuan Gong |
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Institution: | (1) Department of Biological Sciences, National University of Singapore, Singapore 119260, SG |
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Abstract: | Two tissue-specific promoters were used to express both green fluorescent protein (GFP) and red fluorescent protein (RFP)
in transgenic zebrafish embryos. One promoter (CK), derived from a cytokeratin gene, is active specifically in skin epithelia
in embryos, and the other promoter (MLC) from a muscle-specific gene encodes a myosin light chain 2 polypeptide. When the
2 promoters drove the 2 reporter genes to express in the same embryos, both genes were faithfully expressed in the respective
tissues, skin or muscle. When the 2 fluorescent proteins were expressed in the same skin or muscle cells under the same promoter,
GFP fluorescence appeared earlier than RFP fluorescence in both skin and muscle tissues, probably owing to a higher detection
sensitivity of GFP. However, RFP appeared to be more stable as its fluorescence steadily increased during development. Finally,
F1 transgenic offspring were obtained expressing GFP in skin cells under the CK promoter and RFP in muscle cells under the MLC
promoter. Our study demonstrates the feasibility of monitoring expression of multiple genes in different tissues in the same
transgenic organism. |
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Keywords: | : dsRed EGFP cytokeratin myosin light chain skin muscle |
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