Construction of recombinant adenovirus vector carrying human TIMP-1 cDNA and its expression in vitro / 生物医学工程学杂志
Journal of Biomedical Engineering
; (6): 420-424, 2007.
Article
Dans Zh
| WPRIM
| ID: wpr-357685
Responsable en Bibliothèque :
WPRO
ABSTRACT
The full-length cDNA of hTIMP-1 was obtained from a surgical patient with HCC by the method of RT-PCR. Then it was cloned into the adenoviral shuttle plasmid pAdTrack-CMV, and subsequently cotransformed into competent BJ5183 cells with the adenoviral backbone plasmid pAdEasy-1. Thereupon, a recombinant adenoviral plasmid containing full-length cDNA of hTIMP-1 was generated by homologous recombination in E. coli. The adenoviruses (AdhTIMP-1) were packaged and amplified in adenoviral packaging cells HEK 293. Then the viral titer was checked by green fluorescent protein (GFP), and the expression of hTIMP-1 was detected by the techniques of Western blot and RT-PCR. The recombinant adenovirus vector carrying human TIMP-1 was successfully constructed and expressed in vitro and may pave the way for further application in liver gene therapy.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Protéines de fusion recombinantes
/
Lignée cellulaire
/
Adenoviridae
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Clonage moléculaire
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ADN complémentaire
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Carcinome hépatocellulaire
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Inhibiteur tissulaire de métalloprotéinase-1
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Matrice extracellulaire
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Vecteurs génétiques
/
Génétique
Limites du sujet:
Humans
langue:
Zh
Texte intégral:
Journal of Biomedical Engineering
Année:
2007
Type:
Article