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Activity of the medaka translation elongation factor 1α‐A promoter examined using the GFP gene as a reporter
Author(s) -
Kinoshita Masato,
Kani Shuichi,
Ozato Kenjiro,
Wakamatsu Yuko
Publication year - 2000
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1046/j.1440-169x.2000.00530.x
Subject(s) - biology , transgene , green fluorescent protein , microbiology and biotechnology , microinjection , gene , reporter gene , gene isoform , embryo , genetically modified mouse , promoter , eukaryotic translation elongation factor 1 alpha 1 , gene expression , genetics , rna , ribosome
The translation elongation factor 1α (EF‐1α) is known to have several isoforms, which are expressed in a tissue‐ and stage‐specific manner. Two genes encoding EF‐1α exist per haploid genome in the medaka. In the present study, the promoter activity of the 5′‐flanking region of the medaka EF‐1α‐A gene, an isoform of EF‐1α , was characterized using transgenic techniques. First, using CAT gene as a reporter, it was revealed that about 1.8 kbp 5′‐flanking sequence from the transcription initiation site of EF‐1α‐A was sufficient for high‐level promoter activity. Second, the green fluorescent protein ( GFP ) gene fused to this region was introduced into medaka eggs using the microinjection method. Three germline transgenic individuals (one male and two female) were mated with non‐transgenic medaka to obtain F1 offspring. In the case of embryonic and adult F1 transgenic individuals, GFP fluorescence was observed in almost all the tissues examined (e.g. kidney, liver, heart, gill, ovary, and testis), except for the skeletal muscle. In the case of F2 transgenic embryos derived from F1 transgenic males and non‐transgenic females, the fluorescence was observed from the early gastrula stage. On the other hand, in the case of F2 transgenic embryos derived from F1 transgenic females and non‐transgenic males, the fluorescence was observed even at the 1‐cell stage, suggesting that this region is transcriptionally active during oogenesis. The usefulness of the EF‐1α‐A promoter as a tool for introducing foreign proteins into oocytes is discussed.