Characterization of the biological activities of lyophilized recombinant adenovirus expressing the triple mutant of hypoxia inducible factor-1α / 南方医科大学学报
Journal of Southern Medical University
;
(12): 469-472, 2011.
Article
Dans Chinois
| WPRIM
| ID: wpr-307908
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the influence of lyophilization on the biological activity of recombinant adenovirus-mediated triple mutant of hypoxia inducible factor-1α (Ad-HIF-1α-564/402/803).</p><p><b>METHODS</b>Ad-HIF-1α-564/402/803 was amplified from HEK293A cells and purified by ultracentrifugation in CsCl gradient solutions. The infection efficiency was observed by X-gal staining. The lyophilized adenovirus was prepared under appropriate conditions. Before and after lyophilization, the effect of Ad-HIF-1α-564/402/803 on hMVEC proliferation was evaluated by MTS assay. The recombinant adenovirus was confirmed by PCR and DNA sequence analysis before and 1 day, 6 months and 12 months after lyophilization, and hMVECs infected with Ad-HIF-1α-564/402/803 at these time points were examined for HIF-1α protein expression using Western blotting.</p><p><b>RESULTS</b>No significant changes were observed in the effect of lyophilized Ad-HIF-1α-564/402/803 on hMVECs proliferation at the optimal multiplicity of infection of 100 pfu/cell (P>0.05). At the 4 time points, the recombinant adenovirus HIF-1α showed no structural alterations or significant changes in the expression level of HIF-1α protein in the transfected hMVECs (P>0.05).</p><p><b>CONCLUSION</b>Lyophilized Ad-HIF-1α-564/402/803 can maintain its biological activities for a long time.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Adenoviridae
/
Sous-unité alpha du facteur-1 induit par l'hypoxie
/
Protéines mutantes
/
Cellules HEK293
/
Lyophilisation
/
Vecteurs génétiques
/
Génétique
/
Métabolisme
/
Anticorps monoclonaux
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Journal of Southern Medical University
Année:
2011
Type:
Article
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