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The construction of recombinant AAV vector expressing HSVtk gene controlled by Tet-On and the detection of its activity / 生物工程学报
Chinese Journal of Biotechnology ; (12): 360-364, 2005.
Article in Zh | WPRIM | ID: wpr-305269
Responsible library: WPRO
ABSTRACT
In order to investigate the application of recombinant adeno-associated virus (rAAV) vector containing Tet regulation system and HSVtk gene in cancer gene therapy, pAAV/TRE/HSVtk/Tet-On was constructed and identified with PCR and restriction enzyme digestion. Packaging cells HEK293 were cotransfected with plasmids pAAV/TRE/HSVtk/Tet-On, pAAV-RC and pAAV-helper to produce infectious rAAV, and CsCl2 densitygradient centrifugation method was performed for purification and concentration of rAAV. The viruses were then transduced into MCF-7 cells. The results of dot blot hybridization indicate that the rAAV can transfer the target gene into MCF-7 cells. MTT assay showed that GCV could kill AAV-infected MCF-7 cells under the induction of Dox. The data demonstrated that rAAV containing Tet regulation system and HSVtk gene was successfully obtained, and could be used for further investigation of in vivo and in vitro experiments.
Subject(s)
Full text: 1 Index: WPRIM Main subject: Pharmacology / Thymidine Kinase / Transfection / Genetic Therapy / Ganciclovir / Doxycycline / Simplexvirus / Dependovirus / Cell Line, Tumor / Genes, Transgenic, Suicide Type of study: Diagnostic_studies Limits: Humans Language: Zh Journal: Chinese Journal of Biotechnology Year: 2005 Type: Article
Full text: 1 Index: WPRIM Main subject: Pharmacology / Thymidine Kinase / Transfection / Genetic Therapy / Ganciclovir / Doxycycline / Simplexvirus / Dependovirus / Cell Line, Tumor / Genes, Transgenic, Suicide Type of study: Diagnostic_studies Limits: Humans Language: Zh Journal: Chinese Journal of Biotechnology Year: 2005 Type: Article