Expression of FTZ-F1alpha in transgenic Xenopus embryos and oocytes.
Fushi tarazu transcription factor-1 (FTZ-F1) was originally found as a regulator of fushi tarazu gene expression in Drosophila. The frog homologue (FTZ-F1alpha) and the 3.5 kb 5'-flanking region of the FTZ-F1alpha gene have been cloned, and it has been shown by reverse transcription-polymerase chain reaction that FTZ-F1alpha expression begins in embryos at stage 11 and becomes stronger after that. By in situ hybridization analysis, the FTZ-F1alpha mRNA was also found in immature frog oocytes. In this study, immunohistology revealed that the product of FTZ-F1alpha was localized in the cytoplasm of the immature oocyte. To analyze the promoter activity of the Rana rugosa FTZ-F1alpha gene, transgenic Xenopus were produced carrying the fusion construct, consisting of truncated 5'-flanking regions (3.0, 1.8 and 0.3 kb) of the FTZ-F1alpha gene and the green fluorescent protein (GFP) open reading frame. The 0.3 kb 5'-flanking region could drive GFP expression in Xenopus embryos at stage 20 and in immature oocytes in the ovary 2 months after metamorphosis. Gel mobility shift assay was used to test whether proteins in extracts from Xenopus embryos and ovaries bound to the 0.3 kb DNA. The extract from embryos at stage 11 formed one retarded band. The extract from ovaries formed a different retarded band. The results, taken together, indicate that production of transgenic Xenopus is very useful for the analysis of the promoter activity of genes in amphibians. The results also suggest that at least two proteins (one in the embryo and the other in the ovary of 2-month-old postmetamorphosing Xenopus) bind the 0.3 kb 5'-flanking region of the FTZ-F1alpha gene. These proteins may be involved in the regulation of FTZ-F1alpha gene expression in amphibians.
Asahi
Hirotaka
H
Department of Biology, School of Education, Waseda University, 1-6-1 Nishi-Waseda, Shinjuku-ku, Tokyo 169-8050, Japan.
Takase
Minoru
M
Yuge
Masahiro
M
Matsui
Kumi
K
Mori
Mitsuaki
M
Fujita
Takashi
T
Nakamura
Masahisa
M
D024506:5' Flanking Region / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D051717:Fushi Tarazu Transcription Factors; D017930:Genes, Reporter; D005822:Genetic Vectors; D018398:Homeodomain Proteins; D008845:Microinjections; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D018160:Receptors, Cytoplasmic and Nuclear; D054339:Steroidogenic Factor 1; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014981:Xenopus
D004268:DNA-Binding Proteins; D051717:Fushi Tarazu Transcription Factors; D018398:Homeodomain Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D054339:Steroidogenic Factor 1; D014157:Transcription Factors