Open Access

Construction of recombinant pEGFP‑N1‑hPer2 plasmid and its expression in osteosarcoma cells

  • Authors:
    • Anyuan Cheng
    • Yan Zhang
    • Hongjun Mei
    • Shuo Fang
    • Peng Ji
    • Jian Yang
    • Ling Yu
    • Weichun Guo
  • View Affiliations

  • Published online on: March 1, 2016     https://doi.org/10.3892/ol.2016.4291
  • Pages: 2768-2772
  • Copyright: © Cheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of this study was to construct the eukaryotic expression vector pEGFP‑N1‑hPer2 and assess its expression in the human osteosarcoma cell line MG63. Total mRNA was extracted from human osteosarcoma MG63 cells, the human period 2 (hPer2) gene was obtained by reverse transcription‑polymerase chain reaction (RT‑PCR) and cloned into the pEGFP‑N1 vector, then the recombinant pEGFP‑N1‑hPer2 plasmid was constructed and transfected into MG63 cells using Lipofectamine 2000. The expression of hPer2 in MG63 cells was measured by quantitative RT‑PCR and western blot analysis. The accurate construction of pEGFP‑N1‑hPer2 was verified by double enzyme digestion and DNA sequencing. hPer2 gene expression in the transfected cells was assessed by RT‑qPCR and western blot analysis. In conclusion, the recombinant pEGFP‑N1‑hPer2 plasmid was constructed successfully, and expressed effectively in MG63 cells.
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April-2016
Volume 11 Issue 4

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

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Spandidos Publications style
Cheng A, Zhang Y, Mei H, Fang S, Ji P, Yang J, Yu L and Guo W: Construction of recombinant pEGFP‑N1‑hPer2 plasmid and its expression in osteosarcoma cells. Oncol Lett 11: 2768-2772, 2016
APA
Cheng, A., Zhang, Y., Mei, H., Fang, S., Ji, P., Yang, J. ... Guo, W. (2016). Construction of recombinant pEGFP‑N1‑hPer2 plasmid and its expression in osteosarcoma cells. Oncology Letters, 11, 2768-2772. https://doi.org/10.3892/ol.2016.4291
MLA
Cheng, A., Zhang, Y., Mei, H., Fang, S., Ji, P., Yang, J., Yu, L., Guo, W."Construction of recombinant pEGFP‑N1‑hPer2 plasmid and its expression in osteosarcoma cells". Oncology Letters 11.4 (2016): 2768-2772.
Chicago
Cheng, A., Zhang, Y., Mei, H., Fang, S., Ji, P., Yang, J., Yu, L., Guo, W."Construction of recombinant pEGFP‑N1‑hPer2 plasmid and its expression in osteosarcoma cells". Oncology Letters 11, no. 4 (2016): 2768-2772. https://doi.org/10.3892/ol.2016.4291