The stable expression of human tissue-type plasminogen activator gene mediated by lipofectamine in human vein endothelial cell line cells / 生物医学工程学杂志
Journal of Biomedical Engineering
;
(6): 1330-1333, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-230692
ABSTRACT
We have established a human umbilical vein endothelial cell (HUVEC) line monoclonal cells with the stable expression of human tissue-type plasminogen activator (t-PA) gene to provide a basis for further study on the vascular tissue engineering. Recombinant plasmid pcDNA3. 1-Myc-His B (-)/t-PA was constructed by insertion of t-PAcDNA originated from PBS/t-PA into eukaryotic expression vector pcDNA3. 1-Myc-His B(-) and transfected into hUVEC line cells mediated by lipofectamine. The positive clones were obtained by the screen of G418. The transcription and expression of t-PA gene were investigated by RT-PCR and Western blotting respectively. The t-PA activity was measured by chromogenic substrate assay. The positive clone cells which transcripted the mRNA of t-PA gene was obtained by RT-PCR. Immunoreactive human t-PA of the medium was significantly increased in the group of transfected gene when compared with that in the controlled and transfected plasmid without t-PA gene group. The biological activity of the protein of the t-PA in the media was increased significantly in the positive clone cells with t-PA gene transfected. The HUVEC line monoclonal cells with the stable expression of t-PA gene was established successfully.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Recombinant Proteins
/
RNA, Messenger
/
Transfection
/
Cell Line
/
Tissue Plasminogen Activator
/
Tissue Engineering
/
Human Umbilical Vein Endothelial Cells
/
Genetics
/
Lipids
Limits:
Humans
Language:
Chinese
Journal:
Journal of Biomedical Engineering
Year:
2007
Type:
Article
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