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

Cheng,Anyuan; Zhang,Yan; Mei,Hongjun; Fang,Shuo; Ji,Peng; Yang,Jian; Yu,Ling; Guo,Weichun

doi:10.3892/ol.2016.4291

April-2016 Volume 11 Issue 4

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April-2016 Volume 11 Issue 4

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Article 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
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Affiliations: Orthopedic Department, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China, Department of Cardiology, The General Hospital of Fengfeng Group Co., Ltd., Handan, Hebei 056200, P.R. China, Orthopedic Department, Wuhan No. 5 Hospital, Wuhan, Hubei 430050, P.R. China

Copyright: © Cheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pages: 2768-2772
|
Published online on: March 1, 2016

https://doi.org/10.3892/ol.2016.4291
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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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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

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

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Abstract

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