ZFIN ID: ZDB-PUB-050318-14
Cloning and Biological Analysis of the Zebrafish (Danio rerio) Insulin-Like Growth Factor Binding Protein-2 Proximal Promoter Region
Chen, J.Y., Chou, M.J., Gong, H.Y., Huang, T.C., Wu, J.L., and Kuo, C.M.
Date: 2005
Source: DNA and cell biology   24(3): 199-208 (Journal)
Registered Authors: Chen, Jyh-Yih, Gong, Hong-Yi, Wu, Jen-Leih
Keywords: none
MeSH Terms:
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Fireflies/genetics
  • Gene Expression Regulation, Developmental
  • Insulin-Like Growth Factor Binding Protein 2/genetics*
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Transfection
  • Zebrafish/genetics*
PubMed: 15767786 Full text @ DNA Cell Biol.
ABSTRACT
Insulin-like growth factor binding protein 2 (IGFBP-2) plays an important role in the regulation of IGF's action and endocrinology in fish. To understand the molecular mechanism which controls transcription of the IGFBP-2 gene, we cloned and sequenced the IGFBP-2 proximal promoter region of the zebrafish IGFBP-2 gene and characterized its activity by firefly luciferase transient transfection expression assays. Different fragments of the zebrafish IGFBP-2 5'-flanking region were transfected into Hela and ZFL cells. In these cell lines, maximum promoter activity was located in the 900 base pairs (bp) of the zebrafish IGFBP-2 5' flanking region in the ZFL cell line and 318 bp of the zebrafish IGFBP-2 5' flanking region in the Hela cell line. The in vivo actions of the IGFBP-2 promoter on developmental stage expression were further investigated in transgenic zebrafish in which an IGFBP-2 (900-bp) promoter-driven green fluorescent protein encoding the GFP cDNA transgene was microinjected into zebrafish embryos. Morphological and RT-PCR studies of transgenic zebrafish indicated that the IGFBP-2 promoter-driven GFP transcripts appeared for the first time in the 32-cell stage. These results indicate that the IGFBP-2 promoter is active in a development-specific manner. These results suggest that the IGFBP-2 promoter plays an important role in teleost embryo growth.
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