Construction of pEGFP-N1-Fcy::Fur recombinant plasmid and observation of its expression in ovarian cancer cells / 重庆医科大学学报
Journal of Chongqing Medical University
;
(12)2007.
Article
in Chinese
| WPRIM
| ID: wpr-578893
ABSTRACT
Objective:
To construct a EGFP(Enhanced green fluorescent protein)-labled euk-aryotic expression plasmid of FcyFur suicide gene and to detect its expression in SKOV3 cell line.Methods:
With the technology of gene re-arrangement,FcyFur gene in pORF-FcyFur plasmid was subcloned into pEGFP-N1 vector,with its correctness evaluated by the means of r-estriction enzyme analysis and sequencing.It was transfected into SKOV3 cells with lipofectin,the transient expression of GFP was observed under flu- orescence microscope after 24 hours and detected by Western blot.Results:
Correct construction of pEGFP-N1-FcyFur was identi- fied by methods of restriction enzyme analysis and nucleotide sequence determination.A total of 60% transfe-cted cells emitted out green fluorescence under fluorescent microscope after 24 h after transfecti-.on. FcyFur gene expressed by the transfected cells were testified by Western blot.Conclusion:
The recombinant eukaryotic expression vectors have been constructed successfully and effec- tive-ly expressed in ovarian cancer cells,which may provide an experimental basis for gene therapy of ovarian cancer.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Journal of Chongqing Medical University
Year:
2007
Type:
Article
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