Identification of 5′-upstream region of pufferfish ribosomal protein L29 gene as a strong constitutive promoter to drive GFP expression in zebrafish
Journal
Biochemical and Biophysical Research Communications
Journal Volume
314
Journal Issue
1
Pages
249-258
Date Issued
2004
Date
2004
Author(s)
Chang, Ming-Huang
Chou, Chih-Ming
Hsieh, Yueh-Chun
Lu, I-Ching
M.Kothai Nachiar Devi
Chang, Juei-Pei
Huang, Chang-Jen
Abstract
The genomic structure of Tetraodon fluviatilis L29 gene was determined and its promoter activity was analyzed in COS-1 cells and zebrafish embryos. The TfL29 gene comprises four exons and three introns, spanning approximately 1.7kb. The 5′-upstream 2.2-kb of the first exon contains 10 E-boxes and many putative binding motifs for transcription factors GATA-1, AML-1a, c-Myb, Oct-1, CdxA, and NRF-2. Promoter activity assay showed that the distal 2.2-kb fragment not only had high luciferase activity in COS-1 cells, but also strong and ubiquitous GFP expression in a variety of tissues in zebrafish embryos. On the other hand, there are no TATA or CAAT boxes within a 300-bp region upstream from the transcription initiation site. Although this region has high luciferase activity in COS-1 cells, it is not sufficient to drive GFP expression in zebrafish embryos. In this proximal 300-bp region, there are two E-boxes, two CdxA sites, and one NRF-2 site that is immediately downstream of the transcription start site. ? 2003 Elsevier Inc. All rights reserved.
Other Subjects
protein c Myb; ribosome protein; TATA binding protein; transcription factor; transcription factor GATA 1; transcription factor RUNX1; green fluorescent protein; hybrid protein; photoprotein; Rpl29 protein, mouse; animal cell; article; cell strain COS1; controlled study; enzyme activity; exon; fish genetics; gene expression regulation; gene identification; gene location; gene structure; genome analysis; intron; nonhuman; nucleotide sequence; priority journal; promoter region; protein expression; protein motif; puffer fish; transcription initiation site; zebra fish; amino acid sequence; animal; Caenorhabditis elegans; chemistry; comparative study; Drosophila; genetics; human; metabolism; molecular genetics; sequence alignment; sequence analysis; sequence homology; species difference; structure activity relation; Tetraodontiformes; transactivation; Danio rerio; Tetraodon; Tetraodon fluviatilis; Tetraodon fluviatilis; Tetraodontidae; Amino Acid Sequence; Animals; Base Sequence; Caenorhabditis elegans; COS Cells; Drosophila; Gene Expression Regulation; Green Fluorescent Proteins; Humans; Luminescent Proteins; Molecular Sequence Data; Promoter Regions (Genetics); Recombinant Fusion Proteins; Ribosomal Proteins; Sequence Alignment; Sequence Analysis, Protein; Sequence Homology, Amino Acid; Species Specificity; Structure-Activity Relationship; Tetraodontiformes; Trans-Activation (Genetics); Zebrafish
Type
journal article
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