상위피인용논문
- 조회3,023
Abstract
Hae-Chul Parka, 1, Cheol-Hee Kimb, 1, Young-Ki Baec, Sang-Yeob Yeoa, Seok-Hyung Kima, Sung-Kook Honga, Jimann Shina, Kyeong-Won Yooa, Masahiko Hibic, Toshio Hiranoc, Naomasa Mikid, Ajay B Chitnisb, Tae-Lin Huha, 2
a Department of Genetic Engineering, Kyungpook National University, Taegu, 702-701, Korea
b Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, NIH, Bethesda, Maryland, 20892
c Division of Molecular Oncology, Biomedical Research Center, Osaka University School of Medicine, Osaka, 565, Japan
d Biomedical Research Center, Department of Pharmacology, Osaka University School of Medicine, Osaka, 565, Japan
1 Both authors contributed equally to this work.
2 To whom correspondence should be addressed at Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University, Taegu 702-701, Korea.
Abstract
HuC encodes an RNA binding protein homologous to Drosophila elav that serves as an excellent early marker for differentiating neurons. We have characterized the promoter of the zebrafish HuC gene by examining the ability of 5′-upstream fragments to drive expression of green fluorescent protein (GFP) in live embryos. We determined that 2.8 kb of the 5′-flanking sequence is sufficient to restrict GFP gene expression to neurons. The core promoter spans 251 base pairs and contains a CCAAT box and one SP1 sequence but no TATA box is present near the transcription start site. A putative MyT1 binding site and at least 17 E-box sequences are necessary to maintain the neuronal specificity of HuC expression. Interestingly, sequential removal of the putative MyT1 binding site and 14 distal E boxes does not appear to abolish neuronal expression; rather, it leads to a progressive expansion of GFP expression into muscle cells. Further removal of the three proximal E boxes eliminates neuronal and muscle specificity of GFP expression and leads to ubiquitous expression of GFP in the whole body. Identification of key components of the HuC promoter has led to the establishment of a stable zebrafish transgenic line (HuC-GFP) in which GFP is expressed specifically in neurons. We crossed mind bomb (mib) fish with this line to visualize their neurogenic phenotype in live mib-/- mutant embryos. This cross illustrates howHuC-GFP fish could be used in the future to identify and analyze zebrafish mutants with an aberrant pattern of early neurons.
Keywords
HuC; elav; zebrafish; neuron; promoter; Myt1; E box; transgenic; mind bomb.
논문정보
- Research article
- 2000년 11월 (BRIC 등록일 )
- 국내(교신)+국외 연구진
- 의약학 > 응용의학
- 120회 이상 인용된 논문
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