Construction and Quantitative Validation of Chicken CXCR4 Expression Reporter

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• 作者：Masoumeh Es-haghi ; Mohammadreza Bassami ; Hesam Dehghani
• 关键词：Hematopoietic lymphoblast cell line (MSB1) ; Promoter of CXCR4 gene ; Gallus gallus ; Promoter validation
• 刊名：Molecular Biotechnology
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：58
• 期：3
• 页码：202-211
• 全文大小：3,563 KB
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• 作者单位：Masoumeh Es-haghi (1)
  Mohammadreza Bassami (1)
  Hesam Dehghani (1) (2)
  
  1. Division of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran
  2. Embryonic and Stem Cell Biology and Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran
  
• 刊物主题：Biotechnology; Biochemistry, general; Cell Biology; Protein Science; Biological Techniques; Human Genetics;
• 出版者：Springer US
• ISSN：1559-0305

文摘

Site directional migration is an important biological event and an essential behavior for latent migratory cells. A migratory cell maintains its motility, survival, and proliferation abilities by a network of signaling pathways where CXCR4/SDF signaling route plays crucial role for directed homing of a polarized cell. The chicken embryo due to its specific vasculature modality has been used as a valuable model for organogenesis, migration, cancer, and metastasis. In this research, the regulatory regions of chicken CXCR4 gene have been characterized in a chicken hematopoietic lymphoblast cell line (MSB1). A region extending from −2000 bp upstream of CXCR4 gene to +68 after its transcriptional start site, in addition to two other mutant fragments were constructed and cloned in a promoter-less reporter vector. Promoter activity was analyzed by quantitative real-time RT-PCR and flow cytometry techniques. Our findings show that the full sequence from −2000 to +68 bp of CXCR4 regulatory region is required for maximum promoter functionality, while the mutant CXCR4 promoter fragments show a partial promoter activity. The chicken CXCR4 promoter validated in this study could be used for characterization of directed migratory cells in chicken development and disease models.