[Construction of adeno-associated virus vector containing human papilloma virus 16 E7 gene and its expression in eukaryotic cells].
Zhonghua Fu Chan Ke Za Zhi
; 40(7): 480-3, 2005 Jul.
Article
em Zh
| MEDLINE
| ID: mdl-16080877
ABSTRACT
OBJECTIVE:
To construct recombinant adeno-associated virus vector containing human papilloma virus (HPV) 16E7 gene and identify its effectiveness of expression in eukaryotic cell.METHODS:
Recombinant adeno-associated virus particles containing HPV16E7 gene were generated by co-transfection of plasmids pAAV-MCS-E7, pAAV-RC and pAAV-helper into HEK293 cells. The viral particles were collected and purified. The vector titer was measured by southern blot. After the eukaryotic cells were transfected by virus particles, RT-PCR and western blot analysis were performed to identify rAAV expression.RESULTS:
The recombinant adeno-associated virus vector containing HPV16E7 gene was correctly constructed. The vector titer measured by southern blot was approximately 1 x 10(11)/ml. After the eukaryotic cells were transfected by the recombinant adeno-associated virus vector, the expression of HPV16E7 gene was identified by RT-PCR and western blot.CONCLUSIONS:
Recombinant adeno-associated virus HPV16E7 vector is successfully constructed and can stably express in eukaryotic cells.
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Base de dados:
MEDLINE
Assunto principal:
Adenoviridae
/
Papillomavirus Humano 16
/
Proteínas E7 de Papillomavirus
Tipo de estudo:
Risk_factors_studies
Limite:
Female
/
Humans
Idioma:
Zh
Ano de publicação:
2005
Tipo de documento:
Article