title: Construction of recombinant pEGFP-N1-hPer2 plasmid and its expression in osteosarcoma cells.
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.
authors.affiliation: Orthopedic Department, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
authors.forename: Anyuan
authors.initials: A
authors.lastname: Cheng
authors.affiliation: Department of Cardiology, The General Hospital of Fengfeng Group Co., Ltd., Handan, Hebei 056200, P.R. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Orthopedic Department, Wuhan No. 5 Hospital, Wuhan, Hubei 430050, P.R. China.
authors.forename: Hongjun
authors.initials: H
authors.lastname: Mei
authors.affiliation: Orthopedic Department, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Fang
authors.affiliation: Orthopedic Department, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: Ji
authors.affiliation: Orthopedic Department, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Yang
authors.affiliation: Orthopedic Department, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Yu
authors.affiliation: Orthopedic Department, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
authors.forename: Weichun
authors.initials: W
authors.lastname: Guo
keywords: MG63; biological rhythm; circadian gene period 2; eukaryotic expression vector pEGFP-N1; osteosarcoma