Cloning, characterization and promoter analysis of common carp hairy/Enhancer-of-split-related gene, her6.
Some members of hairy/Enhancer-of-split-related gene (HES) family have important effects on axial mesoderm segmentation and the establishment and maintenance of the somite fringe. In fishes, the her6 gene, a member of the HES family, is the homologue of hes1 in mammals and chicken. In this study, the her6 gene and its full-length cDNA from the common carp (Cyprinus carpio) were isolated and characterized. The genomic sequence of common carp her6 is approximately 1.7 kb, with four exons and three introns, and the full-length cDNA of 1314 bp encodes a putative polypeptide of 271 amino acids. To analyse the promoter sequence of common carp her6, sequences of various lengths upstream from the transcription initiation site of her6 were fused to enhanced green fluorescent protein gene (eGFP) and introduced into zebrafish embryos by microinjection to generate transgenic embryos. Our results show that the upstream sequence of 500 bp can direct highly efficient and tissue-specific expression of eGFP in zebrafish embryos, whereas a fragment of 200 bp containing the TATA box and a partial suppressor of hairless paired site sequence (SPS) is not sufficient to drive eGFP expression in zebrafish embryos.
Liu
Jing
J
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Sun
Yong-Hua
YH
Wang
Na
N
Wang
Ya-Ping
YP
Zhu
Zuo-Yan
ZY
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002347:Carps / Q000196:embryology / Q000235:genetics*; D002874:Chromosome Mapping; D002875:Chromosomes; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D004625:Embryo, Nonmammalian; D004742:Enhancer Elements, Genetic; D005091:Exons; D017930:Genes, Reporter; D016678:Genome; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007438:Introns; D008845:Microinjections; D008969:Molecular Sequence Data; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D017386:Sequence Homology, Amino Acid; D024363:Transcription Initiation Site; D015027:Zebrafish / Q000196:embryology / Q000235:genetics
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D012313:RNA; D004247:DNA