American Journal of Life Sciences

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Construction and Functional Analysis of Luciferase Reporter Plasmid Containing CD25 Gene Promoter

Received: Dec. 27, 2017    Accepted:     Published: Dec. 28, 2017
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Abstract

CD25 is the alpha chain of IL-2R and IL-2R is distributed on the surface of activated T cells, B cells, and NK cell. The transcriptional factor CD25 plays an important role in the proliferation and differentiation of Treg cells. MKL-1 is also called MAL (megakaryocytic acute leukemia) and belongs to the Myocardin family which also includes MKL-2 (megakaryoblastic leukemia 2) and Myocardin. Studies have shown that in the process of mouse megakaryocyte differentiation, the expression of MKL-1 is up-regulated, and MKL-1 promotes the physiological maturation of human and mouse megakaryocytes. However, the exact mechanism of the CD25 on megakaryocytes is unclear. In this study, human CD25 promoter luciferase reporter construct was generated by PCR amplification. Then the PCR fragment was digested and cloned into pGL3 vector, and this promoter sequence was verified by sequencing. The result showed that luciferase reporter with CD25 promoter was successfully constructed. The expression vectors of human MKL-1 were bought from addgene. Then the activation of the CD25 promoter was detected in 293T cells by luciferase reporter assay after transfected expression vectors of human MKL-1. The result showed that transfected the expression vectors can enhance the transcriptional activity of CD25. Our research will reveal the effect of CD25 on megakaryocytes and may provide a theoretical basis and therapeutic method for diseases caused by megakaryocytes.

DOI 10.11648/j.ajls.20170506.12
Published in American Journal of Life Sciences ( Volume 5, Issue 6, December 2017 )
Page(s) 160-163
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

CD25, MKL-1, Megakaryocytes, Luciferase Activity Assay

References
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  • APA Style

    Yuan Xiang, Mei Wu, Jia-Peng Li, Zi-Jian Zhang, Zhou-Tong Dai, et al. (2017). Construction and Functional Analysis of Luciferase Reporter Plasmid Containing CD25 Gene Promoter. American Journal of Life Sciences, 5(6), 160-163. https://doi.org/10.11648/j.ajls.20170506.12

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    ACS Style

    Yuan Xiang; Mei Wu; Jia-Peng Li; Zi-Jian Zhang; Zhou-Tong Dai, et al. Construction and Functional Analysis of Luciferase Reporter Plasmid Containing CD25 Gene Promoter. Am. J. Life Sci. 2017, 5(6), 160-163. doi: 10.11648/j.ajls.20170506.12

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    AMA Style

    Yuan Xiang, Mei Wu, Jia-Peng Li, Zi-Jian Zhang, Zhou-Tong Dai, et al. Construction and Functional Analysis of Luciferase Reporter Plasmid Containing CD25 Gene Promoter. Am J Life Sci. 2017;5(6):160-163. doi: 10.11648/j.ajls.20170506.12

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  • @article{10.11648/j.ajls.20170506.12,
      author = {Yuan Xiang and Mei Wu and Jia-Peng Li and Zi-Jian Zhang and Zhou-Tong Dai and Feng Huang and Han-Han Li and Xing-Hua Liao},
      title = {Construction and Functional Analysis of Luciferase Reporter Plasmid Containing CD25 Gene Promoter},
      journal = {American Journal of Life Sciences},
      volume = {5},
      number = {6},
      pages = {160-163},
      doi = {10.11648/j.ajls.20170506.12},
      url = {https://doi.org/10.11648/j.ajls.20170506.12},
      eprint = {https://download.sciencepg.com/pdf/10.11648.j.ajls.20170506.12},
      abstract = {CD25 is the alpha chain of IL-2R and IL-2R is distributed on the surface of activated T cells, B cells, and NK cell. The transcriptional factor CD25 plays an important role in the proliferation and differentiation of Treg cells. MKL-1 is also called MAL (megakaryocytic acute leukemia) and belongs to the Myocardin family which also includes MKL-2 (megakaryoblastic leukemia 2) and Myocardin. Studies have shown that in the process of mouse megakaryocyte differentiation, the expression of MKL-1 is up-regulated, and MKL-1 promotes the physiological maturation of human and mouse megakaryocytes. However, the exact mechanism of the CD25 on megakaryocytes is unclear. In this study, human CD25 promoter luciferase reporter construct was generated by PCR amplification. Then the PCR fragment was digested and cloned into pGL3 vector, and this promoter sequence was verified by sequencing. The result showed that luciferase reporter with CD25 promoter was successfully constructed. The expression vectors of human MKL-1 were bought from addgene. Then the activation of the CD25 promoter was detected in 293T cells by luciferase reporter assay after transfected expression vectors of human MKL-1. The result showed that transfected the expression vectors can enhance the transcriptional activity of CD25. Our research will reveal the effect of CD25 on megakaryocytes and may provide a theoretical basis and therapeutic method for diseases caused by megakaryocytes.},
     year = {2017}
    }
    

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  • TY  - JOUR
    T1  - Construction and Functional Analysis of Luciferase Reporter Plasmid Containing CD25 Gene Promoter
    AU  - Yuan Xiang
    AU  - Mei Wu
    AU  - Jia-Peng Li
    AU  - Zi-Jian Zhang
    AU  - Zhou-Tong Dai
    AU  - Feng Huang
    AU  - Han-Han Li
    AU  - Xing-Hua Liao
    Y1  - 2017/12/28
    PY  - 2017
    N1  - https://doi.org/10.11648/j.ajls.20170506.12
    DO  - 10.11648/j.ajls.20170506.12
    T2  - American Journal of Life Sciences
    JF  - American Journal of Life Sciences
    JO  - American Journal of Life Sciences
    SP  - 160
    EP  - 163
    PB  - Science Publishing Group
    SN  - 2328-5737
    UR  - https://doi.org/10.11648/j.ajls.20170506.12
    AB  - CD25 is the alpha chain of IL-2R and IL-2R is distributed on the surface of activated T cells, B cells, and NK cell. The transcriptional factor CD25 plays an important role in the proliferation and differentiation of Treg cells. MKL-1 is also called MAL (megakaryocytic acute leukemia) and belongs to the Myocardin family which also includes MKL-2 (megakaryoblastic leukemia 2) and Myocardin. Studies have shown that in the process of mouse megakaryocyte differentiation, the expression of MKL-1 is up-regulated, and MKL-1 promotes the physiological maturation of human and mouse megakaryocytes. However, the exact mechanism of the CD25 on megakaryocytes is unclear. In this study, human CD25 promoter luciferase reporter construct was generated by PCR amplification. Then the PCR fragment was digested and cloned into pGL3 vector, and this promoter sequence was verified by sequencing. The result showed that luciferase reporter with CD25 promoter was successfully constructed. The expression vectors of human MKL-1 were bought from addgene. Then the activation of the CD25 promoter was detected in 293T cells by luciferase reporter assay after transfected expression vectors of human MKL-1. The result showed that transfected the expression vectors can enhance the transcriptional activity of CD25. Our research will reveal the effect of CD25 on megakaryocytes and may provide a theoretical basis and therapeutic method for diseases caused by megakaryocytes.
    VL  - 5
    IS  - 6
    ER  - 

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Author Information
  • Department of Molecular Biology, Institute of Biology and Medicine, College of Life and Health Sciences, Wuhan University of Science and Technology, Wuhan, P.R.China

  • Department of Clinical Microbiology, School of Laboratory Medicine, Hubei University of Chinese Medicine, Wuhan, P.R.China

  • Department of Molecular Biology, Institute of Biology and Medicine, College of Life and Health Sciences, Wuhan University of Science and Technology, Wuhan, P.R.China

  • Department of Molecular Biology, Institute of Biology and Medicine, College of Life and Health Sciences, Wuhan University of Science and Technology, Wuhan, P.R.China

  • Department of Molecular Biology, Institute of Biology and Medicine, College of Life and Health Sciences, Wuhan University of Science and Technology, Wuhan, P.R.China

  • Department of Molecular Biology, Institute of Biology and Medicine, College of Life and Health Sciences, Wuhan University of Science and Technology, Wuhan, P.R.China

  • Department of Molecular Biology, Institute of Biology and Medicine, College of Life and Health Sciences, Wuhan University of Science and Technology, Wuhan, P.R.China

  • Department of Molecular Biology, Institute of Biology and Medicine, College of Life and Health Sciences, Wuhan University of Science and Technology, Wuhan, P.R.China

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