PUBLICATION

Epidermis-restricted expression of zebrafish cytokeratin II is controlled by a -141/+85 minimal promoter, and cassette -141/-111 is essential for driving the tissue specificity

Authors
Wang, Y.H., Chen, Y.H., Lu, J.H., Lin, Y.J., Chang, M.Y., and Tsai, H.J.
ID
ZDB-PUB-060517-15
Date
2006
Source
Differentiation; research in biological diversity   74(4): 186-194 (Journal)
Registered Authors
Chen, Yau-Hung, Tsai, Huai-Jen
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • Epidermis/cytology
  • Epidermis/metabolism*
  • Female
  • Gene Expression Regulation*
  • Genes, Reporter
  • Keratin-2/genetics
  • Keratin-2/metabolism*
  • Male
  • Molecular Sequence Data
  • Promoter Regions, Genetic*
  • Recombinant Fusion Proteins/genetics
  • Recombinant Fusion Proteins/metabolism
  • Tissue Distribution
  • Zebrafish/anatomy & histology
  • Zebrafish/metabolism*
PubMed
16683989 Full text @ Differentiation
Abstract
We isolated a 2.3 kb DNA segment from the upstream region of the zebrafish cytokeratin II (zfCKII) gene. Transgenic embryos, produced by using a series of 5' deletions linked to the red fluorescent protein (RFP) reporter, showed that the -141/+85 segment of zfCKII directed RFP expression in epidermal cells, whereas the -111/+85 segment did not. When -141/-111 was deleted from -355/+85 and microinjected into one-celled embryos, no fluorescence was observed at later stages, indicating that the -141/-111 segment is required for green fluorescent protein expression in epidermal cells. Furthermore, when a putative KLF-binding site at -119/-117 was mutated, RFP expression rates and intensities were reduced dramatically, although still observed, suggesting that -119/-117 within -141/-111 is a key cis-element for controlling epidermis-specific expression of the zfCKII gene. Finally, we generated a zebrafish transgenic line, Tg(zfCKII(2.3):RFP), which carries an upstream 2.3 kb regulatory region of the zfCKII gene fused with RFP. The expression pattern in the epidermal cells of Tg(zfCKII(2.3):RFP) fish recapitulated that of the endogenous gene. F2 embryos derived from Tg(zfCKII(2.3):RFP) males crossed with wild-type females revealed that the earliest onset of RFP expression was at the sphere stage, indicating that this transgenic approach can be used for studying zygotic expression of maternally inherited genes.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes