Determination of a necdin cis-acting element required for neuron specific expression by using zebra fish.
To determine cis-acting elements required for neuron specific expression of a necdin gene, we tried to use zebra fish assay system in vivo instead of cell lines in vitro. Various expression vectors carrying upstream sequences of necdin gene fused to MEKA (lacZ) gene as a reporter were injected into fertilized zebra fish embryos and then the expression of the reporter gene was analyzed by the whole mount immunochemical method. No promoter activity was obtained with a construct carrying sequence from -63 to +63 of the necdin gene, while promoter activity with preferential skin expression was obtained with a construct having sequence from -86 to +28. Further upstream sequence from -173 to +28 exhibited neuron specific expression as well as that from -845 to +63. These results indicate that a cis-acting element responsible for neuron specific expression is located in an 87bp sequence from -173 to -87 of necdin gene.
Kuo
C H
CH
Department of Pharmacology I, Medical School, Osaka, Japan.
Uetsuki
T
T
Kim
C H
CH
Tanaka
H
H
Li
B S
BS
Taira
E
E
Higuchi
H
H
Okamoto
H
H
Yoshikawa
K
K
Miki
N
N
D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D017931:DNA Primers; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005136:Eye Proteins / Q000096:biosynthesis*; D020555:GTP-Binding Protein Regulators; D015870:Gene Expression; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics*; D009474:Neurons / Q000378:metabolism*; D009687:Nuclear Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics*; D010750:Phosphoproteins; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D015027:Zebrafish; D001616:beta-Galactosidase / Q000096:biosynthesis
D017931:DNA Primers; D005136:Eye Proteins; D020555:GTP-Binding Protein Regulators; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; C069766:necdin; C065631:phosducin; D001616:beta-Galactosidase