The construction of bicistronic eukaryotic vector carrying green fluorescent protein and hytk gene and its expression in bladder carcinoma / 中国病理生理杂志
Chinese Journal of Pathophysiology
;
(12)1999.
Article
Dans Chinois
| WPRIM
| ID: wpr-518135
ABSTRACT
AIM:
To facilitate the suicide gene delivery into neoplasm, a chimeric gene of HSV-tk and green fluorescent protein (gfp) was constructed.METHODS:
Molecular cloning technique was used to construct this kind of eukaryotic vector. The internal ribosome entry site (IRES) of encephalomyocarditis virus (EMCV), which could coordinate expression of two genes in a single vector, was optioned. By using liposome-mediated transfection, eukaryotic expression vector tgCMV/hytk-IRES-gfp was transfected into human bladder carcinoma cells EJ.RESULTS:
A bicistronic eukaryotic vector carrying gfp and hygromycin phosphotransferase-thymidine kinase fusion (hytk) gene was constructed. The results of PCR and microscopy detection show that the hytk-IRES-gfp gene was successfully transferred into EJ cells. There were no differences in the growth pattern or the morphology between EJ and EJ/hytk-GFP cells. In vitro experiments demonstrated dose- and time-dependent cell killing by transduction of the hytk-IRES-gfp gene followed by GCV treatment. The IC50 (the concentration required to elicit 50% growth inhibition) was 2.16 mg/L in treatment with GCV for 72 hours.CONCLUSION:
These results suggest that this new kind of eukaryotic vector could serves as a new tool and method for neoplasm gene therapy.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
langue:
Chinois
Texte intégral:
Chinese Journal of Pathophysiology
Année:
1999
Type:
Article
Documents relatifs à ce sujet
MEDLINE
...
LILACS
LIS