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.
journal_name
Oncol Lettjournal_title
Oncology lettersauthors
Cheng A,Zhang Y,Mei H,Fang S,Ji P,Yang J,Yu L,Guo Wdoi
10.3892/ol.2016.4291subject
Has Abstractpub_date
2016-04-01 00:00:00pages
2768-2772issue
4eissn
1792-1074issn
1792-1082pii
OL-0-0-4291journal_volume
11pub_type
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