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

文献类型: 外文期刊

作者: Ma, Qian 1 ; Fan, Yanjun 1 ; Zhuang, Zhimeng 1 ; Liu, Shufang 1 ;

作者机构: 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China

2.Qingdao Natl Lab Marine Sci & Technol, Funct Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266200, Peoples R China

3.Qingdao Natl Lab Marine Sci & Technol, Funct Lab Marine Biol & Biotechnolgy, Qingdao 266200, Peoples R China

关键词: cloning;gene expression pattern;promoter transcriptional activity;bone morphogenetic protein;Cynoglossus semilaevis;early developmental stages

期刊名称:ACTA OCEANOLOGICA SINICA ( 影响因子:1.431; 五年影响因子:1.445 )

ISSN: 0253-505X

年卷期: 2018 年 37 卷 2 期

页码:

收录情况: SCI

摘要: 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.

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