Construction of a recombinant adenovirus carrying endostatin gene / 南方医科大学学报
Journal of Southern Medical University
;
(12): 1514-1516, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-283094
ABSTRACT
<p><b>OBJECTIVE</b>To construct a recombinant adenovirus carrying human endostatin gene with AdEasy system.</p><p><b>METHODS</b>Endostatin gene fragment was amplified from Pshuttle-Endostatin plasmid with PCR and subcloned into the pAdTrack-CMV shuttle vector. The resultant plasimid was cotransduced into E.coli BJ 5183 cells with pAdEasy-1 plasmid for homologous recombination. The linearized recombinant plasmid was subsequently transfected into AAV 293 cells, and the recombinant adenovirus was detected by GFP, PCR and restriction analysis.</p><p><b>RESULTS AND CONCLUSION</b>The positive clones of the recombinants were verified by restriction analysis and the titer of the virus reached 2.06 x 10(10)pfu/ml, suggesting successful construction of recombinant adenovirus pAd-Endo.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Genetic Engineering
/
Gene Expression
/
Cell Line
/
Adenoviridae
/
Cloning, Molecular
/
Endostatins
/
Escherichia coli
/
Genetic Vectors
/
Genetics
/
Metabolism
Limits:
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2007
Type:
Article
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