Construction of a fusion expression plasmid containing the G250 gene and human granulocyte-macrophage colony stimulating factor and its significance in renal cell carcinoma

  • Authors:
    • Jun-Wu Li
    • Jun-Yuan Gong
    • Ke Li
    • Yan Liu
    • Qiu-Ping Ye
    • Xin Liu
    • Ze-Xuan Su
  • View Affiliations

  • Published online on: December 30, 2010     https://doi.org/10.3892/ol.2010.230
  • Pages: 343-347
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Abstract

This study aimed to construct a eukaryotic expression plasmid containing the G250/MN/CA IX (G250) and human granulocyte-macrophage colony stimulating factor (hGM-CSF) genes, and to detect the expression of these proteins in vitro by recombinant plasmids in eukaryotic cells. pORF-hGM-CSF and pcDNA3.0-G250 were used as the template to amplify G250 and hGM-CSF by routine polymerase chain reaction (PCR). The two PCR products were cloned into the eukaryotic vector pVAX1, in order to construct a recombinant plasmid pVAX1-G250-hGM, and the plasmid was transfected into human embryonic kidney 293 cells. The protein expression was then determined by immunocytochemi- stry, atomic force microscopy, ELISA and Western blotting. DNA sequencing showed that the cloned G250 and hGM-CSF sequences were consistent with the reported Gene Bank ones. Moreover, a high expression was noted following recombinant plasmid transfection of the G250 and hGM-CSF proteins. Thus, the eukaryotic expression vector pVAX1-G250-hGM containing G250 and hGM-CSF was constructed, allowing for the investigation of the anti-G250 antigen vaccine and immune response mechanisms of biological immunotherapy in renal cell carcinoma.
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March-April 2011
Volume 2 Issue 2

Print ISSN: 1792-1074
Online ISSN:1792-1082

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Spandidos Publications style
Li J, Gong J, Li K, Liu Y, Ye Q, Liu X and Su Z: Construction of a fusion expression plasmid containing the G250 gene and human granulocyte-macrophage colony stimulating factor and its significance in renal cell carcinoma. Oncol Lett 2: 343-347, 2011
APA
Li, J., Gong, J., Li, K., Liu, Y., Ye, Q., Liu, X., & Su, Z. (2011). Construction of a fusion expression plasmid containing the G250 gene and human granulocyte-macrophage colony stimulating factor and its significance in renal cell carcinoma. Oncology Letters, 2, 343-347. https://doi.org/10.3892/ol.2010.230
MLA
Li, J., Gong, J., Li, K., Liu, Y., Ye, Q., Liu, X., Su, Z."Construction of a fusion expression plasmid containing the G250 gene and human granulocyte-macrophage colony stimulating factor and its significance in renal cell carcinoma". Oncology Letters 2.2 (2011): 343-347.
Chicago
Li, J., Gong, J., Li, K., Liu, Y., Ye, Q., Liu, X., Su, Z."Construction of a fusion expression plasmid containing the G250 gene and human granulocyte-macrophage colony stimulating factor and its significance in renal cell carcinoma". Oncology Letters 2, no. 2 (2011): 343-347. https://doi.org/10.3892/ol.2010.230