The construction of recombinant adenovirus expressing bifunctional fusion protein sCAR-EGF and the detection of its activity / 生物工程学报
Chinese Journal of Biotechnology
; (12): 713-719, 2006.
Article
in Zh
| WPRIM
| ID: wpr-286221
Responsible library:
WPRO
ABSTRACT
To improve the targeting of adenovirus vector for gene therapy, a fusion gene sCAR-EGF, in which epidermal growth factor gene was fused to the 3' end of extracellular Coxsackie virus-adenovirus receptor gene, was constructed and cloned into shuttle plasmid pDC315 to obtain a recombinant plasmid pDC315-sCAR-EGF. With the AdMax system, AD-293 cells were co-transfected with pDC315-sCAR-EGF and adenovirus genomic plasmid pBHGloxdeltaE13cre. Through high efficiency site specific recombination, a replication-defective adenovirus Ad5-CMV-sCAR-EGF was constructed. The recombinant adenovirus was analyzed by PCR and Western blotting, the results indicated that Ad5-CMV-sCAR-EGF contained the fusion gene sCAR-EGF, and the adenovirus infected cells was induced to produce and secrete the fusion protein into the supernatant. We have demonstrated that the fusion protein sCAR-EGF is helpful for elevating the infection efficiency of Ad5-CMV-luc with the reporter gene in vitro, which providing a new approach to the gene therapy for tumors overexpressing EGFR.
Full text:
1
Database:
WPRIM
Main subject:
Receptors, Virus
/
Therapeutics
/
Recombinant Fusion Proteins
/
Enzyme-Linked Immunosorbent Assay
/
Genetic Therapy
/
Cell Line
/
Adenoviridae
/
Polymerase Chain Reaction
/
Epidermal Growth Factor
/
Coxsackie and Adenovirus Receptor-Like Membrane Protein
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
Zh
Journal:
Chinese Journal of Biotechnology
Year:
2006
Document type:
Article