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Article

Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk

  • Authors:
    • Eui-Man Jung
    • Beum-Soo An
    • Yu-Kyung Kim
    • Inho Hwang
    • Jong-Yun Lee
    • Tae‑Young Shin
    • Sang-Hwan Hyun
    • Woo-Suk Hwang
    • Eui-Bae Jeung
  • View Affiliations / Copyright

    Affiliations: Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 361-763, Republic of Korea, Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 361-763, Republic of Korea, Sooam Biotech Research Foundation, Seoul 152‑904, Republic of Korea, Laboratory of Veterinary Embryology and Biotechnology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 361-763, Republic of Korea
  • Pages: 406-412
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    Published online on: November 13, 2012
       https://doi.org/10.3892/mmr.2012.1182
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Abstract

Human interferon α (IFN-α) and erythropoietin (EPO) have been used for a variety of purposes in clinical medicine. Human IFN-α has been used to treat several types of viral infection and cancer, as well as renal anemia, via stimulation of erythrocyte formation in the bone marrow. Transgenic cattle are excellent candidates for pharmaceutical production for humans due to their ability to produce recombinant proteins in milk. The purpose of the present study was to generate bovine transgenic fibroblasts capable of producing recombinant human IFN-α and EPO proteins in transgenic cattle milk. First, we analyzed the promoter activities of various bovine milk protein genes in HC11 mouse mammary epithelial cells. The bovine milk protein gene promoters were cloned into the Luc gene in a promoter-less pGL3-Basic vector. Presence of the αS1-casein promoter (-175 to +796 nt) resulted in an up to 16-fold increase in luciferase activity compared with that of the promoter-less construct. In addition, the human IFN-α and EPO genes were identified as significantly overexpressed in HC11 cells compared with the promoter-less construct. Together, the present results demonstrate that the construct with the αS1-casein promoter may induce secretion of recombinant human IFN-α and EPO into bovine milk. Furthermore, we generated transgenic fibroblasts expressing human IFN-α and EPO cDNA controlled by the αS1-casein promoter and two screening markers, enhanced green fluorescent protein and neomycin resistance. These transgenic fibroblasts may be a source of somatic cells for generating transgenic cattle that produce recombinant human IFN-α and EPO proteins during lactation.
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Copy and paste a formatted citation
Spandidos Publications style
Jung E, An B, Kim Y, Hwang I, Lee J, Shin TY, Hyun S, Hwang W and Jeung E: Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk. Mol Med Rep 7: 406-412, 2013.
APA
Jung, E., An, B., Kim, Y., Hwang, I., Lee, J., Shin, T. ... Jeung, E. (2013). Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk. Molecular Medicine Reports, 7, 406-412. https://doi.org/10.3892/mmr.2012.1182
MLA
Jung, E., An, B., Kim, Y., Hwang, I., Lee, J., Shin, T., Hyun, S., Hwang, W., Jeung, E."Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk". Molecular Medicine Reports 7.2 (2013): 406-412.
Chicago
Jung, E., An, B., Kim, Y., Hwang, I., Lee, J., Shin, T., Hyun, S., Hwang, W., Jeung, E."Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk". Molecular Medicine Reports 7, no. 2 (2013): 406-412. https://doi.org/10.3892/mmr.2012.1182
Copy and paste a formatted citation
x
Spandidos Publications style
Jung E, An B, Kim Y, Hwang I, Lee J, Shin TY, Hyun S, Hwang W and Jeung E: Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk. Mol Med Rep 7: 406-412, 2013.
APA
Jung, E., An, B., Kim, Y., Hwang, I., Lee, J., Shin, T. ... Jeung, E. (2013). Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk. Molecular Medicine Reports, 7, 406-412. https://doi.org/10.3892/mmr.2012.1182
MLA
Jung, E., An, B., Kim, Y., Hwang, I., Lee, J., Shin, T., Hyun, S., Hwang, W., Jeung, E."Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk". Molecular Medicine Reports 7.2 (2013): 406-412.
Chicago
Jung, E., An, B., Kim, Y., Hwang, I., Lee, J., Shin, T., Hyun, S., Hwang, W., Jeung, E."Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk". Molecular Medicine Reports 7, no. 2 (2013): 406-412. https://doi.org/10.3892/mmr.2012.1182
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