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Immuno-Localization of DEAD Family Proteins in Germ Line Cells of Xenopus Embryos.
Authors:Masanao Watanabe  Keiji Itoh  Kuniya Abe  Toshifumi Akizawa  Kohji Ikenishi  Mitsuru Furusawa
Affiliation:Furusawa MorphoGene Project, Exploratory Research for Advanced Technology (ERATO), Research Development Corporation of Japan (JRDC), Tsukuba 300-26, Japan;Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Setsunan University, Hirakata 573-01, Japan;Department of Biology, Faculty of Science, Osaka City University, Osaka 558, Japan
Abstract:In order to investigate whether a vasa -like protein is present in germ line cells of Xenopus , antibodies were produced which react specifically with synthetic oligopeptides of sequences from near the N- or C-termini or with one including the DEAD box of the Drosophila vasa protein.
Only the antibody against the oligopeptide including the DEAD box reacted strongly with germ plasm (GP) or with cytoplasm of germ line cells of Xenopus embryos by immunofluorescence microscopy. By immunoelectron microscopy, the antibody was demonstrated to react with the GP-specific structure, germinal granules, in cleaving embryos, and with their derivatives in the germ line cells of embryos at stages extending from gastrula to feeding tadpole. It also reacted with mitochondria not only in the GP and the germ line cells but also in somatic cells, and with myofibrils in muscle cells. By Western blotting, the antibody was shown to react with several bands of Mr 42–69 ± 103 in protein samples from Xenopus embryos. In samples from Drosophila ovaries, it reacted with a Mr 71 ± 103 band which was probably the vasa protein. This indicates the possibility that Xenopus embryos contain several DEAD family proteins. One of these is present on germinal granules, resembling the vasa protein on polar granules of Drosophila .
Keywords:(DEAD protein/vasa-like    germ line cells    germinal granules    Xenopus
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