PUBLICATION
Upstream Regulatory Region of Zebrafish lunatic fringe: Isolation and Promoter Analysis
- Authors
- Liu, J., Sun, Y.H., Wang, N., Wang, Y.P., and Zhu, Z.Y.
- ID
- ZDB-PUB-060616-8
- Date
- 2006
- Source
- Marine biotechnology (New York, N.Y.) 8(4): 357-365 (Journal)
- Registered Authors
- Liu, Jing, Sun, Yonghua
- Keywords
- Green fluorescent protein, lfng, maternal factor, promoter, zebrafish
- MeSH Terms
-
- Animals, Genetically Modified
- Cloning, Molecular
- Blotting, Western
- Gene Expression Regulation, Developmental/physiology
- DNA, Complementary/chemistry
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/physiology
- Animals
- Regulatory Sequences, Nucleic Acid/genetics*
- Sequence Alignment
- DNA Primers/chemistry
- Promoter Regions, Genetic/genetics*
- Zebrafish/genetics*
- Microscopy, Fluorescence
- Molecular Sequence Data
- Green Fluorescent Proteins/analysis
- Green Fluorescent Proteins/biosynthesis
- Glycosyltransferases/genetics*
- Glycosyltransferases/physiology
- Gene Transfer Techniques
- Female
- Base Sequence
- Reverse Transcriptase Polymerase Chain Reaction
- PubMed
- 16729214 Full text @ Mar. Biotechnol.
Citation
Liu, J., Sun, Y.H., Wang, N., Wang, Y.P., and Zhu, Z.Y. (2006) Upstream Regulatory Region of Zebrafish lunatic fringe: Isolation and Promoter Analysis. Marine biotechnology (New York, N.Y.). 8(4):357-365.
Abstract
Lunatic fringe (Lfng), one modulator of Notch signaling, plays an essential part in demarcation of tissues boundaries during animal early development, especially somitogenesis. To characterize the promoter of zebrafish lfng and generate somite-specific transgenic zebrafish, we isolated the upstream regulatory region of zebrafish lfng by blast search at the Ensembl genome database (http://www.ensembl.org) and analyzed the promoter activity using green fluorescent protein (GFP) as a reporter. Promoter activity assay in zebrafish shows that the 0.2-kb fragment containing GC-box, CAAT-box, and TATA-box can direct tissue-specific GFP expression, while the 0.4-kb and 1.2-kb fragments with further upstream sequence included drive GFP expression more efficiently. We produced lfngEGFP-transgenic founders showing somite-specific expression of GFP and consequently generated a hemizygous lfngEGFP-transgenic line. The eggs from lfngEGFP-transgenic female zebrafish show strong GFP expression, which is consistent to the reverse-transcription polymerase chain reaction PCR (RT-PCR) detection of lfng transcripts in the fertilized eggs. This reveals that zebrafish lfng is a maternal factor existing in matured eggs, suggesting that fish somitogenesis may be influenced by maternal factors.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping