title: Upstream regulatory region of zebrafish lunatic fringe: isolation and promoter analysis.
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.
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.forename: Yong-Hua
authors.initials: YH
authors.lastname: Sun
authors.forename: Na
authors.initials: N
authors.lastname: Wang
authors.forename: Ya-Ping
authors.initials: YP
authors.lastname: Wang
authors.forename: Zuo-Yan
authors.initials: ZY
authors.lastname: Zhu
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000737:chemistry; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D018014:Gene Transfer Techniques; D016695:Glycosyltransferases / Q000235:genetics* / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000096:biosynthesis; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D015027:Zebrafish / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics* / Q000502:physiology
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins; D016695:Glycosyltransferases; C498390:lfng protein, zebrafish