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Acta Oceanologica Sinica

, Volume 37, Issue 2, pp 76–84 | Cite as

Cloning, expression profiling and promoter functional analysis of bone morphogenetic protein 2 in the tongue sole (Cynoglossus semilaevis)

  • Qian Ma
  • Yanjun Fan
  • Zhimeng Zhuang
  • Shufang Liu
Article
  • 19 Downloads

Abstract

BMP2 plays crucial roles in vertebrate developmental process and acts as a bone inducer during osteogenesis. We present here the molecular cloning of bmp2 cDNA from the marine flatfish Cynoglossus semilaevis, and the analysis of bmp2 expression profiling and promoter function. The full length of bmp2 cDNA sequence is 2 048 bp, which encodes a protein of 422 amino acids. Tissue expression distribution of bmp2 was examined in 14 tissues of mature individuals by quantitative real time PCR (qRT-PCR). The results revealed that bmp2 was expressed ubiquitously, and the highest expression level was detected in the spinal cord. Moreover, bmp2 expression levels were detected at 15 sampling time points of early developmental stages (egg, larva, juvenile and fingerling stages). The highest expression level of bmp2 was observed at the gastrula stage, which was about ten times higher than those at the other three embryo stages. Whole-mount in situ hybridization showed that the bmp2 signal was strongly detected at the location of the crown-like larval fin, heart and liver, and slightly expressed in the notochord at one day post hatch (dph); then the expression of bmp2 started to be concentrated in notochord at three dph. Subsequently, we characterized the 5′-flanking region of bmp2 by testing the promoter activity by Luciferase reporter assays. Positive regulatory region was detected at the location of–179 to +109. The predicted transcription factor binding sites (E-box binding factors, zinc finger transcription factor, etc.) in this region might participate in the transcriptional regulation of the bmp2 gene.

Keywords

cloning gene expression pattern promoter transcriptional activity bone morphogenetic protein Cynoglossus semilaevis early developmental stages 

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Copyright information

© The Chinese Society of Oceanography and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Qian Ma
    • 1
    • 2
  • Yanjun Fan
    • 1
  • Zhimeng Zhuang
    • 1
    • 3
  • Shufang Liu
    • 1
    • 2
  1. 1.Yellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdaoChina
  2. 2.Function Laboratory for Marine Fisheries Science and Food Production ProcessesQingdao National Laboratory for Marine Science and TechnologyQingdaoChina
  3. 3.Function Laboratory for Marine Biology and BiotechnolgyQingdao National Laboratory for Marine Science and TechnologyQingdaoChina

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