Construction of eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG and its expression in vitro
- VernacularTitle:人真核表达质粒载体pcDNA3.1-hOPG的构建及体外表达
- Author:
Hua TANG
;
Ping GONG
;
Zhen TAN
;
Dapeng LIAO
- Publication Type:Journal Article
- Keywords:
osteoprotegerin;
eukaryotic expression plasmid
- From:Journal of Third Military Medical University
2003;0(23):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG,and to detect the expression of the plasmid in vitro.Methods hOPG cDNA fragment was extracted from plasmid MGC:29565,amplified by PCR,and inserted into pcDNA3.1(-)vector,then identified by restriction endonuclease digestion and nucleotide sequencing.C2C12 cells were transfected with the plasmid using Lipofectamine 2000.The expression of OPG protein was detected by immunohistochemistry techniques and bone wafer pit assay.Results The constructed recombinant plasmid contained the sequence of hOPG gene.After transfection with the plasmid,active OPG protein could be expressed in C2C12 cells.Conclusion The constructed eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG can express OPG in vitro,which is the basis for further study on the treatment of bone absorption caused by abnormal activity of osteoclasts.
